Effects of rumen-protected methionine supplementation and bacterial lipopolysaccharide infusion on nitrogen metabolism and hormonal responses of growing beef steers

Metabolic demand for sulfur-containing AA increases during inflammation in nonruminants. Therefore, Met supplementation may alleviate the negative effects of infection on N balance. Effects of gram-negative bacterial lipopolysaccharide (LPS) and supplemental dietary Met on N balance, serum hormones and haptoglobin, and plasma urea-N and AA were evaluated in 20 Angus-cross steers (BW = 262 +/- 6.3 kg). Treatments (2 x 2 factorial) were infusion of no LPS (-LPS) or a prolonged low dose of LPS (+LPS) and dietary supplementation of no (-MET) or 14 g/d (+MET) of rumen-protected Met (providing 7.9 g/d of dl-Met). Steers were adapted to a roughage-based diet (DMI = 1.4% of BW daily) and supplemental Met for 14 d, and were then infused (1 mL/min via intravenous catheter) with LPS on d 1 (2 microg/kg of BW) and 3 (1 microg/kg of BW) of a 5-d collection period. Blood was collected on d 1, before LPS infusion, and at 2, 4, 6, 8, 10, 12, and 24 h after LPS challenge. Diet samples, feed refusals, feces, and urine were collected daily for 5 d. Rectal temperature and serum concentrations of cortisol, prolactin, tumor necrosis factor-alpha, and haptoglobin increased, whereas thyroxine and triiodothyronine decreased for +LPS vs. -LPS steers (LPS x h; P < 0.01). Plasma urea-N was greater for +LPS than -LPS steers (LPS; P = 0.03), and serum IGF-1 was not affected (P > or = 0.26) by LPS or Met. Plasma concentrations of Thr, Lys, Leu, Ile, Phe, Trp, Asn, Glu, and Orn decreased, plasma Ala increased, and Gly and Ser initially increased, then declined in +LPS vs. -LPS steers (LPS x h; P < or = 0.04). Plasma Met was greater for +MET than -MET steers before LPS infusion, but declined in +MET steers after LPS infusion (LPS x Met x h; P < 0.01). By design, DMI was not different, but DM digested was less (P = 0.04) for +LPS than -LPS steers. Infusion of LPS did not affect (P > or = 0.24) N intake, fecal N excretion, or N digested, but resulted in greater (P < 0.01) urinary N excretion and less (P < 0.01) N retention. The absence of an LPS x Met interaction (P = 0.26) for N retention indicates that supplemental Met does not improve the N utilization of growing beef steers exposed to a gram-negative bacterial endotoxin. Decreases in plasma concentrations of several essential AA in +LPS steers suggest that metabolic demand for these AA likely increased in steers exposed to endotoxin.     

Modeling the effects of estradiol and progesterone on the acute phase proinflammatory axis: variability in tumor necrosis factor-α, nitric oxide, and xanthine oxidase responses to endotoxin challenge in steers

The severity of host response in some diseases differs between sexes, and this dimorphism has been attributed to the immunomodulating effects of reproductive steroid hormones. In females, susceptibility to disease stress has been associated with reproductive status and attributed to prevailing progesterone (P4) or estrogen concentrations during different estrous cycle phases. Our objective was to clarify and define the effect of P4 or 17β-estradiol (E2) on the acute proinflammatory component of the innate immune system by administering these hormones to steers and evaluating initial and tolerance-associated concentration patterns of circulating proinflammatory immune response mediators after two consecutive lipopolysaccharide (LPS) challenges (LPS1 and LPS2, 6 d apart; 2.5 μg/kg BW, intravenously, Escherichia coli 055:B5). Plasma concentrations of the proinflammatory initiation cytokine tumor necrosis factor-α (TNF-α), nitrate+nitrite [NO(x), estimate of nitric oxide (NO) production], haptoglobin (HG; acute phase protein) and plasma xanthine oxidase activity (mediator of superoxide production) were measured. Crossbred steers (392 ± 7 kg) were fed a forage-concentrate diet (15% CP) to appetite and assigned to control (C; n = 7), P4 (n = 8), or E2 (n = 5) treatment. Jugular blood samples were obtained at 0, 1, 2, 3, 4, 7, and 24 h relative to each of the two LPS injections. For each proinflammatory biomarker, the area under the time by concentration curve (AUC) was used to evaluate and compare responses to the LPS challenge. Treatment with E2 disrupted LPS tolerance as observed in augmented plasma TNF-α (P < 0.01) and NO(x) (P < 0.01) responses to LPS2. Compared with C, P4 treatment decreased plasma NO(x) AUC after LPS2 (P < 0.05) and tended to reduce TNF-α AUC after LPS1 (P = 0.08). Plasma xanthine oxidase activity AUC was increased (P < 0.01) over C by E2 treatment after both LPS1 and LPS2. HG response to LPS1 within 24 h was not affected by any treatment. However, 6 d after LPS1 plasma HG concentration remained higher (P < 0.01) in steers treated with E2 than with C or P4. Results indicate that in cattle, P4 and E2, respectively, attenuate or amplify the response to LPS challenge at several points critical to the regulation of the progression of the proinflammatory cascade.     

The effect of abrupt weaning of suckler calves on the plasma concentrations of cortisol, catecholamines, leukocytes, acute-phase proteins and in vitro interferon-gamma production

The objective of this study was to examine the effect of abrupt weaning (inclusive of social group disruption and maternal separation) on the physiological mediators of stress and measures of immune function. Thirty-six male and 36 female calves (Limousin and Charolais crosses), habituated to handling, were blocked by sex, weight, and breed of dam and randomly assigned, within block, to either a control or abruptly weaned group. Animals were separated into the respective treatment groups at 0 h. Calves were bled at -168, 6 (males only), 24, 48, and 168 h after weaning, and the behavioral reaction of calves to handling was scored. Cortisol, catecholamine (not sampled at -168 h), acute-phase protein concentrations, and in vitro interferon-gamma production and neutrophil:lymphocyte ratio were measured. The effects of weaning, calf sex, time, and their respective interactions were described. Disruption of the established social group at 0 h increased (P < 0.001) the plasma cortisol concentration and neutrophil:lymphocyte ratio and decreased the leukocyte concentration (P < 0.001) and the in vitro interferon-gamma response to the mitogen concanavalin-A (P < 0.001) and keyhole limpet hemocyanin (P < 0.001) for weaned and control animals compared with -168 h. There was no effect of weaning or sex on the behavioral reaction of calves to handling. Plasma cortisol and adrenaline concentrations were not affected by weaning or sex. Plasma noradrenaline concentration was influenced by weaning x sex (P < 0.05) and time x sex (P < 0.05). The response increased for male calves with weaning and increased with each sampling time after weaning. For heifers, the response was not affected by weaning and plasma concentrations decreased at 168 h after weaning. There was no effect of weaning or sex on leukocyte concentration. The neutrophil:lymphocyte ratio increased after weaning (P < 0.01) and was affected by sex (P < 0.05). Weaning decreased (P < 0.05) the in vitro interferon-gamma response to the antigen keyhole limpet hemocyanin. There was a time x weaning x sex (P < 0.05) interaction for fibrinogen concentration but no effect of treatment on haptoglobin concentration. Abrupt weaning increased plasma cortisol and noradrenaline concentrations that were accompanied by attenuation of in vitro interferon-gamma production to novel mitogen and antigen complexes up to 7 d after weaning.     

Serum immunoglobulin concentrations in genetically different types of suckling beef calves in a tropical environment

Factors influencing the serum concentrations of gamma-globulin (gamma-G) during the neonatal period were studied in Shorthorn x Hereford (SH), Africander x SH and Brahman x SH calves born to cows grazing in a tropical environment. There were no significant effects of age of dam, sex or breed of calf on the gamma-G concentrations of calves from birth to 48 hours of age. Concentrations of gamma-G fell within two ranges: group A, 10 to 20 g/l and group B, 35 to 70 g/l. The number of calves in each group was not significantly different between breeds and overall 30% of calves were in group A. Body weight gain from birth to 10-day-old was greater (P less than 0.01) in calves in group B than in group A. Plasma cholesterol concentrations in 10-day-old calves were higher in group B than in group A calves supporting the interpretation that group B calves had higher milk intakes. Identification of calves receiving adequate amounts of colostrum has fundamental significance for the efficient production of cattle in the tropics.     

Tumor necrosis factor as a potential mediator of acute metabolic and hormonal responses to endotoxemia in calves

The effects of coliform endotoxin (E) and recombinant bovine tumor necrosis factor alpha (TNF) were compared with respect to clinical signs of disease and changes in plasma metabolite and pituitary and pancreatic hormone concentrations in calves. In addition, changes in plasma TNF concentration during each challenge exposure were quantitated by use of radioimmunoassay. Healthy Holstein bull calves with mean body weight of 90 kg were each given, in order, on different days, saline solution (5.0 ml, IV, day 1, n = 4), E (type 055:B5, 1.0 micrograms/kg of body weight IV, day 2, n = 4) and TNF (5.0 micrograms/kg IV, day 9, n = 3). Jugular venous blood samples, rectal temperature reading, and PCV were obtained at hourly intervals before (2 hours) and after challenge exposure. The PCV increased (P less than 0.05) after E and TNF administrations for the first 5 hours, then returned to normal in calves given E, but decreased and remained low in calves given TNF through 24 hours. Plasma triglyceride and nonesterified free fatty acids concentrations were increased through 10 hours (P less than 0.05) after E administration, whereas triglyceride and nonesterified free fatty acids concentrations were not significantly affected by TNF administration. Increase in blood glucose concentration at 1 hour after administration of E and TNF was followed by prolonged hypoglycemia that lasted through 6 hours. Changes in plasma insulin concentration paralleled the observed changes in glucose concentration, initially increased at 2 hours after E and TNF (P less than 0.05) administrations, but then tended to decrease below control values thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of transportation and commingling on the acute-phase protein response,growth, and feed intake of newly weaned beef calves

The objective of this study was to investigate the effect of transportation and commingling on measures of the acute-phase protein response in newly weaned beef calves. Thirty-two (Exp. 1; average BW = 266 +/- 20.8 kg) and thirty-six (Exp. 2; average BW = 222 +/- 34.6 kg) Brahman-crossbred calves were randomly allotted to one of four treatments (2 x 2 factorial arrangement [transportation x commingling] in a completely randomized design). Body weight and jugular blood were collected at weaning, after shipment, and 1, 3, and 7 d after transport for Exp. 1, and at weaning and 1, 5, 9, 13, 17, and 21 d after transport for Exp. 2. Feed intake within pen was recorded daily for Exp. 2. Plasma fibrinogen, ceruloplasmin, haptoglobin, and cortisol concentrations were determined for all collection times. Additionally, serum amyloid-A and alpha-acid glycoprotein concentrations were determined in Exp. 1 and 2, respectively. In Exp. 2, commingled calves tended (P = 0.13) to have a higher DMI than noncommingled calves (5.3 and 4.8 kg/d, respectively). Transported calves lost more BW than nontransported calves from the time of weaning to d 1 (2.0 and 3.1% more BW loss for Exp. 1 and 2, respectively). With the exception of haptoglobin in Exp. 1, each of the acute-phase proteins measured in these studies increased over each sampling day. In Exp. 1, transported calves had higher (P < 0.05) mean serum amyloid-A concentrations than nontransported calves (48.9 vs. 33.4 microg/mL). There was a significant sampling day x transportation interaction (P < 0.01) for fibrinogen, ceruloplasmin, and haptoglobin in Exp. 1; transported calves had higher concentrations of fibrinogen following transport and on d 2 and 3, and ceruloplasmin on d 3. Haptoglobin concentrations were higher (P = 0.04) in nontransported calves on d 1 and 2 of Exp. 1. In Exp. 2, overall mean haptoglobin concentrations were higher in nontransported vs. transported calves. The results of these studies indicate that stressors associated with transportation affect the acute-phase protein response in newly weaned beef calves. More research is needed to determine whether these proteins might be valuable indicators of stress following the weaning process.     

Integrated adrenal, somatotropic, and immune responses of growing pigs to treatment with lipopolysaccharide

The objective of this research was to provide an integrated look at systemic adrenal, somatotropic, and immune responses of growing pigs to challenge with lipopolysaccharide (LPS). Weaned pigs were challenged intraperitoneally with 100 microg/kg BW of LPS or sterile saline, and rectal temperature and blood data were collected for 72 h. Daily feed intake also was monitored. Plasma was analyzed for concentrations of cortisol, tumor necrosis factor alpha (TNFalpha), the acute phase protein haptoglobin, growth hormone (GH), insulin-like growth factor I (IGF-I), and prostaglandin E2 (PGE2). As expected, LPS decreased feed intake, stimulated a febrile response, and activated the hypothalamic-pituitary-adrenal (HPA) axis as demonstrated by increased cortisol levels. Cortisol reached maximum elevation 2 h after treatment (P < .001) and remained elevated through 12 h (P < .001). Circulating TNFalpha was increased by LPS at 2 and 4 h after treatment (P < .001), and an apparent (not statistically significant) increase in haptoglobin also occurred in challenged animals. The LPS injection suppressed IGF-I by 2 h following treatment (P < .01), and circulating IGF-I remained reduced relative to controls through 44 h. Overall, GH was increased in LPS-treated pigs (P < .05), although the treatment x time interaction was not significant. Plasma PGE2 was increased transiently at 2 h (P < .05) and then subsequently suppressed at 4, 8, and 12 h following LPS (P < .05). This study provides a comprehensive view of systemic effects of LPS on components of the HPA, growth, and immune axes. In addition, these are the first data to document changes in circulating PGE2 in unrestrained animals during the early hours of the acute phase response to LPS.     

Influence of orally administered bovine lactoferrin on lipid metabolism in lipopolysaccharide-injected preruminant calves

The aim of this study was to investigate the influence of oral lactoferrin (LF) administration on lipid metabolism changes in calves given lipopolysaccharide (LPS). Twenty-one 4-day-old Holstein calves were divided into three groups, with each group receiving one of three oral doses of LF (0, 1, 3 g/day) for 10 consecutive days (day −10 to day −1). All calves were intravenously injected with LPS (50 ng/kg BW) on day 0, the day after LF treatment ended. Plasma triglyceride concentrations were lower ( P  < 0.05) in the LF-treated calves than in the control calves given 0 g/day of LF at 12 and 24 h after LPS injection. Plasma NEFA concentrations were elevated between 6 and 24 h after LPS treatment. At 12 h, the concentration of plasma NEFA was lower ( P  < 0.05) in the calves given LF 3 g/day than in the control calves. On day 0, plasma total cholesterol and phospholipid concentrations tended to be lower in the LF groups administered 1 and 3 g of LF/day than in the control group, but did not differ significantly among the groups. The plasma very-low-density and low-density lipoprotein concentrations were lower ( P  < 0.05) at 12, 24, and 72 h in the LF groups than in the control calves. The concentrations of plasma high-density lipoprotein tended to be lower in the LF groups than in the control group between day 0 and 96 h, though there were no significant group differences. The concentration of plasma interleukin-1β was lower ( P  < 0.05) in the calves fed LF 3 g/day than in the control calves at 2 and 12–48 h after LPS injection. These data suggest that LF inhibits LPS-induced alterations in lipid metabolism in preruminant calves.     

Effects of feeding beef females supplemental fat during gestation on cold tolerance in newborn calves.

Effects of prepartum fat supplementation of the dam on cold tolerance of calves were determined in two studies. In Exp. 1, 22 F1, crossbred heifers gestating F2 calves received diets containing either 1.7 or 4.7% dietary fat starting at d 230+/-2d of gestation. Safflower seeds (Carthamus tinctorius) containing 37% oil with 79% linoleic acid were the supplemental fat source in isocaloric-isonitrogenous diets. Calves were separated from their dams at birth, fed pooled dairy-cow colostrum, muzzled to prevent sucking, and returned to their dams in a heated (22 degrees C) barn for 3.5 h. At 4 h of age, a jugular catheter was inserted. At 5 h of age, calves were placed in a 0 degrees C room for 140 min and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for glucose, cortisol, and cholesterol. In Exp. 2, 18 multiparous, crossbred beef cows bred to Murray Grey sires were randomly assigned to receive diets containing either 1.7 or 3.1% dietary fat starting at 235+/-2 d gestation. Safflower seeds were used as the supplemental fat source in isocaloric-isonitrogenous diets. At d 260 of gestation, premature parturition was induced in one-half of the cows from each diet group by feeding Ponderosa pine (Pinus ponderosa) needles. Experimental protocols were the same as in Exp. 1, except that cold exposure was at 9 degrees C for 200 min. Rectal temperatures were affected in Exp. 1 by time and diet x time (both P < .01) and diet x calf sex (P < .05) and in Exp. 2 by calf age (P < .05), time, and calf age x time (both P < .01). Plasma cortisol concentrations were affected by time (P < .01) and calf sex x time (P < .05) in Exp. 1 and by time ( P < .01) in Exp. 2. Cholesterol concentrations in Exp. 1 were affected by diet x time (P < .05) and in Exp. 2 by time (P < .05). Plasma glucose concentrations were affected in Exp. 1 by diet (P < .05) and in Exp. 2 by calf age, time, and calf age x time (all P < .01). We conclude from Exp. 1 that feeding heifers supplemental fat during late gestation increased glucose concentrations in the newborn calf, resulting in favorable responses in body temperature in the cold-stressed newborns. This increase in substrate availability suggests a potential positive effect on heat generation in newborns during sustained periods of cold stress. In Exp. 2, premature calves had compromised cold tolerance possibly due to impaired shivering or brown adipose tissue thermogenesis.     

Alterations in clinical, hematological and metabolic variables in bovine neonatal endotoxemia.

Endotoxemia is an important cause of morbidity and mortality in the neonate. Although many models are used to study the problem, none completely simulates the natural disease. To more clearly define a bovine neonatal endotoxemia model we studied the effects of dose of endotoxin on clinical, hematological and biochemical variables. Thirty-four neonatal calves were administered Escherichia coli endotoxin (LPS) at 0 (0.9% saline solution), 0.2, 2.0 or 20 micrograms/kg, by either IV bolus or infusion over 50 minutes. Variables monitored included mean arterial blood pressure (MAP), leukocyte (WBC) count, plasma glucose and lactate concentrations and clinical status. All LPS-treated calves displayed similar clinical signs within one hour. Dose-dependent differences in response to LPS among groups became evident over time. Substantial dose-dependent changes in attitude, appetite, mucous membrane character, capillary refill time, MAP, plasma glucose and lactate concentrations, and WBC count were noted in LPS-treated calves. Higher doses of LPS induced a more prolonged clinical response and significantly (p < 0.05) greater hypotension, lacticemia and hypoglycemia. While dose altered the response to endotoxin, the method of administration had no overall effect on the variables measured.     

Effect of growth and parturition on hair cortisol in Holstein cattle

The effect of growth and parturition on hair cortisol concentrations of cattle was investigated. Plasma, saliva, and hair (black and white from the shoulders and hip) samples were collected from calves at 6 and 24 weeks old and from dairy cattle at the dry (1 and 2 months prepartum) and lactation (10, 50, 150, and 250 days postpartum) periods. Plasma and saliva cortisol concentrations were lower in 24-week-old calves than those of 6-week-old calves, and hair cortisol concentrations decreased regardless of color and position. In 6-week-old calves, hair cortisol concentrations differed between sampling positions, but this difference was not observed in 24-week-old calves. Plasma and saliva cortisol concentrations increased before parturition until 10 days postpartum then decreased until 50 days postpartum. The same trend was observed in the cortisol concentrations of white hair. Contrarily, cortisol concentrations in black hair remained unchanged and was lower than that in white hair. Hair cortisol concentration can vary greatly depending on the location on the body, hair color, cattle age, or parturition. When this method is used, all of the above factors must be considered.     

Effects of prepartum supplementary fat and muscle hypertrophy genotype on cold tolerance in newborn calves.

Effects of feeding pregnant dams supplemental dietary fat during the last 55 d of gestation on cold tolerance of newborn crossbred calves with (Piedmontese cross, P, n = 15) or without (Hereford cross, H, n = 16) the muscle hypertrophy allele was determined. Primiparous F1 dams gestating F2 calves of the respective breeds were assigned randomly within breed to receive gestation diets containing either 2.2 (Low Fat; LF) or 5.1% fat (High Fat; HF). Safflower (Carthamus tinctorius L.) seeds containing 37% oil with 79% linoleic acid were the supplemental fat source in diets formulated to be isocaloric-isonitrogenous. At parturition, calves were separated from their dams, fed 38 degrees C pooled dairy cow colostrum (30 mL/kg BW), muzzled to prevent suckling, and returned to their dams in a heated (22 degrees C) room for 3.5 h. At 4 h of age (birth = 0 h), a catheter was inserted into the jugular vein. At 5 h of age, calves were placed in a 0 degrees C room for 140 min, and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for cortisol and glucose. Rectal temperature was affected by diet (P<.05), time, diet x time, and breed x time (P<.01 for time and the interactions). Cortisol and glucose concentrations were not affected by diet, breed, or the diet x breed interaction, but they were affected by time, breed x time (both P<.01), and diet x time (P = .06). Calves from HF dams had higher rectal temperatures than calves from LF dams, and the HF calves maintained higher rectal temperatures throughout cold exposure. Cortisol concentrations were lower (P = .06) in calves from HF dams, and these calves had more (P = .06) glucose available for metabolic heat production than calves from LF dams. Piedmontese-cross calves maintained higher (P<.01) rectal temperatures and had higher cortisol and glucose (both P<.01) concentrations than did H-cross calves. We conclude that feeding dams supplemental fat during late gestation increased heat production in newborn calves and potentially could increase calf survival; calves with muscle hypertrophy may have a different ratio of shivering vs nonshivering thermogenesis due to differences in body composition or relationships among uncoupling proteins.     

Effect of bovine lactoferrin feeding on lipopolysaccharide-induced metabolic and hormonal disturbances in preruminant calves

One of the biological functions of bovine lactoferrin (LF) is modulation of the host defense system, including cytokine production and immune response. The aim of the present study was to investigate the effect of oral administration of LF in calves on lipopolysaccharide (LPS)‐induced metabolic and hormonal changes in inflammatory response. Thirty Holstein calves at 4 day of age were given one of three oral doses of LF (0, 1, 3 g/day) for 10 days (?10 day to ?1 day). They were injected i.v. with LPS (50 ng/kg bodyweight) the day (day 0) after the end of LF treatment. Plasma samples were obtained on ?10, 0 day (immediately before LPS injection), and at 2, 6, 12, 24, 48, 72, and 96 h after LPS injection. Plasma tumor necrosis factor‐α concentrations at 2 h after LPS treatment were lower (P < 0.05) in LF 1 g/day‐fed claves compared with LF 0 g/day (control) calves. On day 0 there were no significant group differences in plasma LF concentration. Plasma concentration of haptoglobin in control calves was elevated by LPS injection. In LF groups, plasma haptoglobin concentrations slightly increased after LPS injection, but those levels at 6–24 h were lower (P < 0.05) than in the control group. The LF treatment inhibited (P < 0.05) the reduction of plasma ferrin concentration in calves following LPS challenge. The concentration of plasma aspartate aminotransferase in calves treated with LF was lower (P < 0.05) than in control calves at 24–96 h after LPS treatment. The concentration of plasma insulin‐like growth factor‐1 (IGF‐1) in all groups was decreased by LPS treatment, while in the LF groups the IGF‐1 level was higher (P < 0.05) than in the control group. Plasma adrenocorticotropic hormone and insulin concentrations in LF groups were lower (P < 0.05) than in control calves at 2 h after LPS injection. These data suggest that LF has a substantial anti‐inflammatory effect on the modulation of the host defense system in preruminant calves.     

Immunohistochemical localization of Pasteurella haemolytica A1-derived endotoxin, leukotoxin, and capsular polysaccharide in experimental bovine Pasteurella pneumonia

Six, 5- to 10-week-old male Holstein calves were inoculated intratracheally with 5 x 10(9) logarithmic growth phase Pasteurella haemolytica biotype A serotype 1 (A1). Immunohistochemical techniques in conjunction with the use of monoclonal antibodies directed against P. haemolytica A1-derived lipopolysaccharide (LPS), capsular polysaccharide, and a polyclonal rabbit anti-leukotoxin antibody were used to localize their respective antigens in tissue sections of pneumonic lung at the light and electron microscopic levels. We found the following: 1) LPS, capsular polysaccharide, and leukotoxin were released into the inflammatory exudate; 2) LPS was found within the cytoplasm of neutrophils (located in the alveolus and alveolar wall), alveolar macrophages, endothelial cells, pulmonary intravascular macrophages, and on epithelial cell surfaces; 3) capsular polysaccharide was found in the alveolus and alveolar macrophages but not in cells of the alveolar wall; and 4) leukotoxin was associated with cell membranes of degenerating inflammatory cells located in the alveolus. This is the first study that demonstrates the presence of leukotoxin in the pulmonary inflammatory lesions caused by P. haemolytica A1 and implicates endotoxin as an important factor in the genesis of the pulmonary lesions.     

Serum concentrations of tumor necrosis factor-alpha in neonatal calves with presumed septicemia

This study was performed to determine the concentrations of tumor necrosis factor (TNF) in the serum of neonatal calves with presumed sepsis and determine the correlation between serum concentrations of TNF and the severity and outcome of disease. Thirty-five sick calves < 30 days old that suffered from enteritis, respiratory disease, or both were considered suitable for inclusion in this study by satisfying clinical and laboratory criteria suggestive of septicemia. At admission, blood samples were collected from all calves to determine the prevalence of high concentrations of TNF. The clinical course and outcome of disease then were recorded. Of the 35 calves with presumed sepsis, 10 had high serum TNF concentrations. Scleral injection, weak or absent suckling reflex, sternal or lateral recumbency, unresponsive or comatose state, and death rate of calves with high serum TNF concentration were greater than those values for calves without high serum TNF concentration. Calves with high serum TNF concentration had significantly lower mean IgG (P < .001), globulin (P < .0001), and calcium (P < .0001) concentrations; greater serum creatinine concentrations (P < .0001); and > or = 2+ toxic changes in neutrophils than did calves without high serum TNF concentrations. Mean values for packed cell volume, band neutrophil count, and venous Pco2 were significantly (P < .007) higher in the group of calves with high serum TNF concentration. Results of this study indicate that serum TNF concentration is correlated with clinical criteria of sepsis in neonatal calves. A close association was apparent between disease severity and serum TNF concentrations in this group of calves with presumed septicemia.     

Endotoxemia and Tumor Necrosis Factor Activity in Dogs With Naturally Occurring Parvoviral Enteritis

A prospective, nonrandomized study was performed to evaluate the role of endotoxin and tumor necrosis factor (TNF) in dogs with parvoviral enteritis. Seventeen dogs with naturally occurring parvoviral enteritis were enrolled in the study. Plasma samples were obtained for quantification of endotoxin and TNF on presentation and at 3 and 6 hours after therapy with either fluids prior to antibiotics, or fluids concurrently with antibiotics. All dogs received standard supportive therapy. Fourteen of 17 dogs had endotoxin in their plasma during the study period; 7 of 17 dogs had measurable TNF. No endotoxin or TNF was detectable in plasma from normal puppies. An increase in TNF activity was pre dictive of mortality ( P = .041). There was a trend for increasing endotoxin activity to predict mortality ( P = .0769). Animals that received antibiotics with fluids were significantly older than those that received fluids prior to antibiotics, and there was a trend for animals that received antibiotics with fluids to have a decrease in endotoxin activity after treatment ( P = .054). Endotoxin and activation of the cytokine cascade are integral to the pathophysiology of parvoviral enteritis. Measures to limit endotoxemia and the systemic inflammatory response may improve survival.     

Effects of intravenous infusion of lipopolysaccharide on plasma micromineral, magnesium, and cytokine concentrations and serum cortisol concentrations in lactating goats

OBJECTIVE: To assess the effects of various doses of lipopolysaccharide (LPS) administered IV on plasma microminerals, magnesium, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6 concentrations and serum cortisol concentrations in lactating goats. ANIMALS: 6 lactating goats. PROCEDURES: Goats were allotted to 3 LPS-treatment groups: control (0 microg/kg), low LPS (10 microg/kg), and high LPS (50 microg/kg). Rectal temperatures and behaviors of goats were recorded immediately before a 10-minute IV infusion of LPS and at 0.5, 1, 2, 4, 6, 8, and 24 hours after infusion. Blood samples were obtained before IV infusion and at 0.5, 1, 2, 4, 6, 8, and 24 hours after infusion. Plasma zinc, copper, iron, and magnesium concentrations were determined by atomic absorption spectrometry; plasma TNF-alpha and IL-6 concentrations were measured by use of an ELISA; and serum cortisol concentrations were determined by use of a radioimmunoassay. RESULTS: A monophasic fever developed in low-LPS and high-LPS groups. In the low-LPS and high-LPS group, plasma zinc concentrations decreased at 6 hours after infusion; compared with control groups. Plasma iron concentrations were lower at 24 hours after infusion in low-LPS and high-LPS groups than in the control group. Plasma TNF-alpha and IL-6 concentrations were higher in low-LPS and high-LPS groups than in the control group at 1, 2, and 4 hours after infusion. In low-LPS and high-LPS groups, serum cortisol concentrations increased from 0.5 hours onward and peaked at 1 (high-LPS group) and 2 (low-LPS group) hours after infusion. CONCLUSIONS AND CLINICAL RELEVANCE: Following IV infusion of LPS, the immune system is activated, which might affect micromineral homeostatic regulation and, subsequently, the metabolic health of lactating goats.     

Effects of nonsteroidal anti-inflammatory drugs on long-term pain in calves castrated by use of an external clamping technique following epidural anesthesia

OBJECTIVE: To compare efficacy of flunixin meglumine versus carprofen in controlling pain under field conditions following castration by use of an external clamping technique in calves that received epidural anesthesia. ANIMALS: 40 male 5- to 6-month-old calves. PROCEDURES: Calves were allocated to 4 groups: castrated only (control calves; n=8); castrated 5 minutes after epidural injection of 2% lidocaine (epidural-alone treated calves; 8), castrated after epidural anesthesia and s.c. administration of flunixin meglumine (epidural-flunixin treated calves; 12), and castrated after epidural anesthesia and s.c. administration of carprofen (epidural-carprofen-treated calves; 11 [1 calf not included]). Plasma cortisol concentration was measured before and 6, 24, and 48 hours after castration. Time of arrival at the feed trough at 24 and 48 hours was observed. Calves were observed at 24 and 48 hours for 4 pain-related behaviors. RESULTS: At 6 hours, control calves had significantly higher plasma cortisol concentrations, compared with baseline values and those of epidural-flunixin- and epidural-carprofen-treated calves. At 24 hours, epidural-carprofen-treated calves had significantly lower plasma cortisol concentrations, compared with control calves. At 48 hours, epidural-carprofen-treated calves had plasma cortisol concentrations that were similar to baseline values and significantly lower than epidural-flunixin- and epidural-alone-treated calves. At 24 and 48 hours, epidural-carprofen-treated calves were first to arrive at the feed trough and had fewer pain-related behaviors. CONCLUSIONS AND CLINICAL RELEVANCE: s.c. administration of carprofen in combination with epidural injection of lidocaine may improve the welfare of calves castrated by use of an external clamping technique for up to 48 hours.     

Effects of Ostertagia ostertagi infection on secretion of metabolic hormones in calves.

Effects of Ostertagia ostertagi infection on secretion of insulin, pancreatic glucagon, cortisol, gastrin, and pepsinogen were studied in calves inoculated with 100,000 (group 1) or 10,000 (group 2) O ostertagi infective larvae weekly for 14 weeks. Plasma insulin concentrations in both inoculated groups were lower than those in a non-infected (group 3) control group. The differences between group 1 and group 3 were significant (P < 0.05) at 2 and 12 weeks after initial inoculation. Plasma pancreatic glucagon and cortisol concentrations of groups 1 and 2 did not differ significantly from those of the control group, although plasma pancreatic glucagon concentration was consistently lower in group-1 calves from 4 weeks to end of the study. Plasma pepsinogen and serum gastrin concentrations also increased significantly (P < 0.05) in both groups that received inoculations. We concluded that decreased plasma insulin concentrations are contributory to changes in postabsorptive protein metabolism, and that serum gastrin concentrations are more representative of the pathologic changes in the abomasum than are plasma pepsinogen concentrations.     

Plasma concentrations of substance P and cortisol in beef calves after castration or simulated castration

OBJECTIVE: To evaluate plasma concentrations of substance P (SP) and cortisol in calves after castration or simulated castration. ANIMALS: 10 Angus-crossbred calves. PROCEDURES: Calves were acclimated for 5 days, assigned to a block on the basis of scrotal circumference, and randomly assigned to a castrated or simulated-castrated (control) group. Blood samples were collected twice before, at the time of (0 hours), and at several times points after castration or simulated castration. Vocalization and attitude scores were determined at time of castration or simulated castration. Plasma concentrations of SP and cortisol were determined by use of competitive and chemiluminescent enzyme immunoassays, respectively. Data were analyzed by use of repeated-measures analysis with a mixed model. RESULTS: Mean +/- SEM cortisol concentration in castrated calves (78.88+/-10.07 nmol/L) was similar to that in uncastrated control calves (73.01+/-10.07 nmol/L). However, mean SP concentration in castrated calves (506.43+/-38.11 pg/mL) was significantly higher than the concentration in control calves (386.42+/-40.09 pg/mL). Mean cortisol concentration in calves with vocalization scores of 0 was not significantly different from the concentration in calves with vocalization scores of 3. However, calves with vocalization scores of 3 had significantly higher SP concentrations, compared with SP concentrations for calves with vocalization scores of 0. CONCLUSIONS AND CLINICAL RELEVANCE: Similar cortisol concentrations were measured in castrated and control calves. A significant increase in plasma concentrations of SP after castration suggested a likely association with nociception. These results may affect assessment of animal well-being in livestock production systems.