Evaluation of phagocytosis, bactericidal activity, and production of superoxide anion, nitric oxide, and tumor necrosis factor-alpha in Kupffer cells of neonatal pigs.

OBJECTIVE: To evaluate the activity of Kupffer cells (KC) of control neonatal pigs and neonatal pigs treated with endotoxin and to compare activity of KC with that of pulmonary alveolar macrophages (PAM). SAMPLE POPULATION: Kupffer cells and PAM obtained from 24 neonatal pigs (7 to 10 days old). PROCEDURE: Pairs (n = 7) of littermates served as treated (lipopolysaccharide [LPS]) or untreated pigs. Pigs were euthanatized 24 hours after treatment, and cells were isolated. Cells were obtained from 10 other neonatal pigs for other assays. Functional activity of cells was evaluated by use of in vitro assays to evaluate bactericidal activity, phagocytosis, and production of superoxide anion (SOA), nitric oxide (NO), and tumor necrosis factor-alpha (TNF-alpha). Each assay was repeated on cells obtained from 4 to 6 pigs. RESULTS: Phagocytic activity was similar in KC and PAM, but bactericidal activity and production of SDA and TNF-alpha was lower in KC. Neither KC nor PAM produced NO in response to LPS stimulation. Phagocytosis, bactericidal activity, and production of SOA were enhanced for KC obtained from neonatal pigs treated with LPS. The PAM from LPS-treated neonatal pigs had similar bactericidal activity to PAM obtained from untreated pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Functional capacity of KC is affected by endotoxin. This provides additional information of the role the liver plays in immune surveillance. In addition, the response of KC in neonatal pigs exposed to endotoxin is of value for understanding gram-negative bacterial sepsis, which is a major cause of mortality in neonatal pigs.     

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma

The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 μg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5 h. Serum was harvested and stored at −80 °C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-) and interferon-gamma (IFN-γ). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time×Treatment×Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time×Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time×Treatment×Diet interactions were also observed for serum concentrations of TNF- (P=0.084) and IFN-γ (P=0.022) such that both the TNF- and IFN-γ response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=−0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=−0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge.     

Effect of Escherichia coli endotoxin and thyrotropin-releasing hormone on prolactin in lactating sows

Primiparous gilts were given subcutaneous injections of saline solution or 8 mg of Escherichia coli endotoxin (055:B5 strain) in saline solution on postpartum days (PPD) 2 and/or 6 and saline solution at the same site on PPD 1, 3, 5, and 7 at 1000 hours. On PPD 1 to 3 and on PPD 5 to 7, pigs were given 100 micrograms of thyrotropin-releasing hormone (TRH) IV at 1300 hours to evaluate TRH-induced prolactin (PRL) release. Blood samples were analyzed for PRL, cortisol, triiodothyronine (T3), and tetraiodothyronine (T4) concentrations. Rectal temperatures were monitored at hourly intervals between 0800 and 1500 hours on PPD 2 and 6. The PRL declined after endotoxin administration on PPD 2, but a similar decline was not seen after saline solution administration on PPD 1, 2, or 3. The PRL concentrations remained unchanged on PPD 5, 6, and 7 in gilts exposed to endotoxin for the 1st or 2nd time on PPD 6 and to saline solution on PPD 5 and 7. The TRH injection caused increases in PRL in all animals, but the PRL increase after TRH injection was significantly lower (P less than 0.05) in gilts treated with endotoxin on PPD 2. Cortisol concentrations increased after endotoxin exposure on PPD 2 and 6. Rectal temperatures increased after endotoxin exposure on PPD 2 and 6 with peak temperatures of 41.8 C and 41.6 C seen 4 and 3 hours, respectively, after endotoxin injection. The T3 and T4 response, used as an indicator of TRH perfusion of the adenohypophysis, was unchanged after endotoxin or saline solution administration.(ABSTRACT TRUNCATED AT 250 WORDS)     

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 sodium butyrate on growth performance and response to lipopolysaccharide in weanling pigs

Two experiments were conducted to determine the effects of dietary sodium butyrate on growth performance and response to Escherichia coli lipopolysaccharide (LPS) in weanling pigs. In a 28-d experiment, 180 pigs (initial BW 6.3 kg) were fed 0, 0.05, 0.1, 0.2, or 0.4% sodium butyrate, or 110 mg/kg of dietary tylosin. There was no effect of dietary sodium butyrate or tylosin on overall G:F, but there was a linear trend (P < 0.07) toward decreased ADFI and ADG as levels of sodium butyrate increased. In a second 28-d experiment, 108 pigs (initial BW 6.3 kg) were assigned to 1 of 3 dietary treatments: 1) no antibiotics, 2) 0.2% sodium butyrate, or 3) 55 mg/kg of carbadox. On d 14, a subset of pigs from the no-antibiotic and butyrate treatment groups was challenged with E. coli LPS or injected with sterile saline in a 2 x 2 factorial arrangement (+/-LPS challenge; +/-dietary butyrate; n = 6 pigs/treatment group). Four hours after LPS challenge, blood samples were obtained, and samples of LM, liver, and ileum were collected for gene expression analysis. Serum samples were analyzed for IL-6, tumor necrosis factor alpha (TNFalpha), alpha(1)-acid glycoprotein, cortisol, IGF-I, insulin, and metabolites. The relative abundance of tissue cytokine and IGF-I mRNA was measured by real-time PCR. Feeding diets containing sodium butyrate or carbadox did not alter ADG or ADFI compared with pigs fed the control diet. From d 0 to 14, pigs fed diets containing 0.2% sodium butyrate had decreased (P < 0.05) ADG and tended (P < 0.06) to have decreased G:F compared with animals fed diets containing carbadox. Challenge with LPS increased (P < 0.05) serum cytokines and cortisol and decreased (P < 0.05) serum glucose and triglycerides. Injection with LPS increased (P < 0.05) the relative abundance of hepatic IL-6 and TNFalpha mRNA, increased (P < 0.05) LM TNFalpha mRNA content, and decreased (P < 0.05) IGF-I mRNA in LM. For serum cortisol, there was an interaction (P < 0.05) between dietary butyrate and LPS. The increase in serum cortisol attributable to LPS was greater (P < 0.05) in pigs fed butyrate than in pigs fed the control diet. There tended (P < 0.10) to be an interaction between LPS and diet and for butyrate to increase the relative abundance of IL-6 mRNA in LM. Carbadox did not alter cytokine or IGF-I mRNA or serum metabolites, but did decrease (P < 0.05) serum TNFalpha. These data indicate that dietary sodium butyrate does not enhance growth performance, but may regulate the response to inflammatory stimuli in weanling pigs.     

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. II. Effects on the immune axis when fed simple or complex diets containing no spray-dried plasma

A trial using 64 weanling pigs (TR4×PIC C22) was conducted to determine the effects of menhaden fish oil supplementation and diet complexity on performance and immune response of nursery pigs. Pigs (17 days and 6.27±1.16 kg) were weaned into a segregated early wean facility and given free access to a complex diet for 7 days post-weaning. At day 0 (day 7 post-weaning), pigs were blocked by weight and allotted to 64 pens. Treatments (Trt) were arranged as a 2×2×2 factorial arrangement. Main effects included diet (complex versus simple), oil (menhaden fish (MFO) versus corn (CO)), and immunogen (saline versus lipopolysaccharide (LPS)). Experimental diets contained 6% oil (6% CO or 5% MFO+1% CO) and were fed for 14 days. On day 12, i.v. injections of either LPS (150 μg/kg) or saline were given, followed by blood collection at 30 min intervals for 6 h. After the immune challenge (day 14), pigs were placed onto a common corn-soybean meal fortified diet and growth performance was evaluated until termination of the study (day 28). Pigs were weighed and feed intakes recorded at 7, 14, and 28 days. Prior to immune challenge (day 12), there were differences in BW for pigs fed complex versus simple diets (P<0.01; 13.1 and 12.1 kg, respectively) and pigs fed CO versus MFO diets (P<0.05; 12.9 and 12.3 kg, respectively). During the challenge period, for pigs treated with LPS there was a Time×Immunogen×Oil effect (P<0.001) for serum cortisol with MFO fed pigs having lower serum cortisol as compared to CO fed pigs. Also, during the challenge period, for pigs treated with LPS there was a Time×Diet×Immunogen×Oil effect (P<0.001) for serum tumor necrosis factor- (TNF-) with pigs fed complex diets supplemented with CO having higher serum TNF- as compared with pigs fed complex diets supplemented with MFO. At days 14 and 28, LPS-treated pigs had lower BW than saline injected controls (P<0.001 and 0.01, respectively). In addition, pigs fed simplified diets continued to have lower BW after challenge compared to pigs fed a complex diet. Interestingly, there were no differences (P>0.10) in BW after challenge in pigs fed MFO. This study suggests that MFO supplementation alters the immune response during LPS challenge and that simplified diets may compromise nursery performance.     

Flunixin meglumine attenuation of endotoxin-induced damage to the cardiopulmonary vascular endothelium of the pony

Endotoxic shock was induced in 5 ponies by intraperitoneal injections of 20, 40, 60, 80, and 80 micrograms of Escherichia coli endotoxin (LPS)/kg of body weight at 0, 6, 12, 18, and 24 hours, respectively. At 24 hours, the ponies also were given 20 micrograms of LPS/kg via catheter in the left ventricle of the heart. A 2nd group of 4 ponies was given 1.1 mg of flunixin meglumine (FM)/kg, IV, at 6, 12, 18, and 24 hours just before the corresponding LPS injection. Two hours after the 24-hour LPS injection, the ponies in both groups were anesthetized, the lungs were perfused with fixative, and portions of the pulmonary arteries and veins and right and left ventricles were prepared for scanning and transmission electron microscopy. In ponies that were given only LPS, some areas of pulmonary vascular endothelium appeared normal when compared with untreated controls, but other areas had disoriented endothelial cells or had varying amounts of sloughing, which ranged from focal areas of a few cells to large areas of denuded endothelium. Ponies treated with FM before LPS had less severe and less extensive endothelial cell damage. In both groups, leukocytes were attached to areas of the vessel wall; endothelial cell damage was greater in these regions. Administration of FM before LPS administration attenuated the LPS-induced endothelial cell damage.     

Acute feed intake and acute-phase protein responses following a lipopolysaccharide challenge in pigs from two dam lines

This study was conducted to evaluate the response of two dam lines of pigs to acute increases of LPS. Acute-phase proteins were also measured to determine their potential use as biological indicators of the immune response. Thirty-six pigs (initial body weight = 21.3 +/- 0.48 kg) were allotted by dam line (Lines 1 and 2) and sex (castrates and gilts) to one of three LPS dose treatments and penned individually. Treatments were a single i.m. injection of 0 (LPS-0), 25 (LPS-25) or 50 microg LPS/kg body weight (BW) (LPS-50). Acute changes in feed intake were related to a pre-injection baseline intake. Feeders were weighed daily to establish baseline feed intake (average daily feed intake -48 to 0 h prior to injection). The acute feed intake response (AFIR) was computed as the average daily feed intake 0-48 h after injection divided by baseline intake. Serum was harvested at time 0 and 48 h after injection. LPS-0 pigs grew faster and consumed more feed than the LPS-25 or LPS-50 pigs (0.79 kg/d versus 0.51 and 0.50 kg/d; 1.15 kg/d versus 0.96 and 0.89 kg/d, respectively; P<0.001). The AFIR of Line 1 castrates and Line 2 gilts was similar for LPS-25 and LPS-50 treatments, while Line 1 gilts and Line 2 castrates had decreased AFIR with increased LPS dose (sex x line x LPS, P<0.05). Three of 18 castrates died but no gilts died following the LPS challenge (P<0.10). Castrates had higher haptoglobin (Hpt) concentrations than gilts on d 0 (18.1 units of absorption/mg of protein versus 13.1 units of absorption/mg of protein; P<0.03). Line 1 pigs had higher C-reactive protein (CRP) concentrations than Line 2 pigs (P<0.05) on d 0. LPS treatment did not change serum concentrations of CRP, Hpt or ceruloplasmin (Cp). However, the change in serum amyloid A (SAA) concentration decreased quadratically (from 0 to 48 h) with increasing LPS dose (P<0.02). This change in SAA was negatively correlated with the AFIR (r= -0.80; P<0.001). In general, castrates appear to be more sensitive to endotoxin challenges than gilts. Serum amyloid A, but not the other acute-phase proteins evaluated, was a good biological indicator of immune system activation following an acute lipopolysaccharide challenge when compared to the acute change in feed intake.     

Effects of the addition of endotoxin during perfusion of isolated forelimbs of equine cadavers

Objective-To examine the effect of endotoxins on metabolism and histopathologic changes of isolated perfused equine forelimbs. Sample-Forelimbs (comprising the metacarpus and digit) were collected from cadavers of 12 healthy adult horses after slaughter at an abattoir (14 limbs; 1 forelimb of 10 horses and both forelimbs of 2 horses). Procedures-Forelimbs were perfused for 10 hours with autologous blood, with and without the addition of endotoxin (80 ng of lipopolysaccharide [LPS]/L). Two limbs of the endotoxin exposure group and 2 nonperfused limbs were loaded to failure of the suspensory apparatus of the pedal bone to evaluate the effect of body weight. Metabolic and histologic variables were evaluated. Results-Blood pressure increased during the first hour and did not differ between groups. Lactate dehydrogenase activity was similar in both groups and increased significantly during the 10-hour period; glucose consumption at 5 hours and lactate concentration at 8 hours were significantly higher in limbs exposed to endotoxin. The width of secondary epidermal lamellae was greater in LPS limbs. In the primary dermal lamellae of LPS limbs, there were significantly more vessels with an open lumen and aggregates of intravascular neutrophils. Conclusions and Clinical Relevance-In the blood-perfused isolated forelimbs of equine cadavers, exposure to LPS led to significant changes in the laminar tissue as well as to metabolic changes. Therefore, endotoxin should be considered as a causative factor for laminitis and not merely as a risk factor.     

内毒素血症山羊体征变化及氨基胍对其影响

将22只山羊随机分为5组，即对照组、内毒素血症模型组（内毒素LPS1800EU／kg）、内毒素血症氨基胍组（AG25mg／kg）、大剂量内毒素血症模型组（内毒素LPS5400EU／kg）和大剂量内毒素血症氨基胍组。肉眼观察动物精神状态和粪便的变化；用体温计检测直肠体温；用听诊器记录呼吸频率。结果表明：实验后1～9h，大、小剂量的内毒素（LPS）均使山羊表现出不同程度的精神不振、腹泻、食欲废绝。体温显著升高（P〈0．05），24h后体温恢复正常（P〉0．05）；但氨基胍对内毒素血症时体温的变化无调理作用（P〉0．05）。小剂量LPS和氨基胍对呼吸频率无影响（P〉0．05）；大剂量LPS却显著增加了山羊呼吸频率（P〈0．05），同时氨基胍也显著阻止了呼吸频率的升高（P〈0．05）。揭示体温、呼吸与内毒素剂量存在依赖关系；小剂量的氨基胍（25mg／kg）对内毒素血症时体温变化没有影响，但可明显减少呼吸频率。     

Effects of fish oil supplementation on the performance and the immunological, adrenal, and somatotropic responses of weaned pigs after an Escherichia coli lipopolysaccharide challenge

Seventy-two crossbred pigs (7.58 +/- 0.30 kg BW) weaned at 28 +/- 3 d of age were used to investigate the effects of fish oil supplementation on pig performance and on immunological, adrenal, and somatotropic responses following an Escherichia coli lipopolysaccharide (LPS) challenge in a 2 x 2 factorial design. The main factors consisted of diet (7% corn oil [CO] or 7% fish oil [FO]) and immunological challenge (LPS or saline). On d 14 and 21, pigs were injected intraperitoneally with either 200 microg/kg BW of LPS or an equivalent amount of sterile saline. Blood samples were collected 3 h after injection for analysis of interleukin-1beta (IL-1beta), prostaglandin E2 (PGE2), cortisol, growth hormone (GH), and insulin-like growth factor (IGF)-I. On d 2 after LPS challenge, peripheral blood lymphocyte proliferation (PBLP) was determined. Lipopolysaccharide challenge decreased ADG (487 vs. 586 g; P < 0.05) and ADFI (as-fed, 776 vs. 920 g; P < 0.05) from d 14 to 21 and ADG (587 vs. 652 g; P < 0.10) from d 21 to 28. Fish oil improved ADG (554 vs. 520 g; P < 0.10) and ADFI (891 vs. 805 g; P < 0.10) from d 14 to 21. On d 14, LPS challenge x diet interactions were observed for IL-1beta (P < 0.10), PGE2 (P < 0.001), and cortisol (P < 0.05) such that these measurements responded to the LPS challenge to a lesser extent (IL-1beta: 93 vs. 114 pg/mL, P < 0.05; PGE2: 536 vs. 1,285 pg/mL, P < 0.001; cortisol: 143 vs. 206 ng/mL, P < 0.05) in pigs receiving the FO diet than in pigs fed the CO diet. In contrast, among LPS-treated pigs, pigs fed the FO diet had higher IGF-I (155 vs. 101 ng/mL; P < 0.10) than those fed the CO diet. On d 21 among LPS-treated pigs, pigs fed FO had lower IL-1beta (70 vs. 84 pg/mL; P < 0.10) and cortisol (153 vs. 205 ng/mL; P < 0.05) than those fed CO. Pigs fed FO had lower PGE2 (331 vs. 444 pg/mL; P < 0.05) and higher IGF-I (202 vs. 171 ng/mL; P < 0.10) compared with those fed CO. Lipopolysaccharide challenge decreased GH (0.27 vs. 0.33 ng/mL; P < 0.05) on d 14, whereas it had no effect on GH on d 21. During both LPS challenge periods, the challenge increased PBLP when these cells were incubated with 8 (1.46 vs. 1.32; P < 0.10) or 16 microg/mL (1.46 vs. 1.30; P < 0.05) of concanavalin A. Fish oil had no effect on PBLP. These results suggest that FO alters the release of proinflammatory cytokines, which might lead to improved pig performance during an immunological challenge.     

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.     

Endotoxin-induced changes in the hemostatic system in neonatal calves: the effect of antiserum to a mutant Escherichia coli (J-5)

Changes in the hemostatic system were studied in 22 neonatal calves given a small dosage of Escherichia coli endotoxin (0.5 microgram/kg) by slow (5-hour) IV infusion. The effect of pretreatment with an antiserum to mutant of E coli O111:B4 (J-5) was evaluated. The platelet count, plasma fibrinogen concentration, prothrombin time, and activated partial thromboplastin time changed significantly from base line during and after endotoxin infusions in all calves. The mean platelet count was significantly decreased from 1 through 24 hours after endotoxin infusion was started. Mean plasma fibrinogen was decreased 2 through 12 hours after endotoxin infusion was started. The mean prothrombin time and activated partial thromboplastin time were significantly greater than base line at 3 to 6 hours and 3 to 12 hours, respectively, after endotoxin infusion was started. Serum concentration of fibrinolytic degradation products remained less than 10 micrograms/ml. Bovine J-5 antiserum did not prevent the endotoxin-induced changes in the hemostatic system of these neonatal calves.     

Efficacy of flunixin meglumine for the treatment of endotoxin-induced bovine mastitis

The clinical effect of flunixin meglumine administration was determined in cows with acute mastitis induced by intramammary administration of endotoxin. In 12 lactating cows, 10 micrograms of Escherichia coli 026:B6 endotoxin were administered via a teat cannula into the teat cistern of single randomly selected rear quarters. Cows were challenge exposed as pairs. One cow in each pair was administered parenteral flunixin meglumine (6 cows) and 1 cow per pair was administered saline solution (6 cows). Multiple doses (7) of 1.1 mg of flunixin meglumine/kg of body weight or saline solution were administered at 8-hour intervals beginning 2 hours after endotoxin. Cow and quarter clinical signs as well as milk somatic cell concentrations, bovine serum albumin, electrical conductivity, and milk production were determined before and for 14 days after endotoxin inoculation. Intramammary endotoxin produced signs characteristic of acute coliform mastitis. Quarter and systemic abnormalities occurred and milk production was reduced by approximately 50% at 12 hours after endotoxin. Flunixin meglumine therapy significantly (P less than or equal to 0.05) reduced rectal temperatures and quarter signs of inflammation and improved clinically graded depression when compared with these signs in saline solution-treated controls. Milk production and laboratory indicators of inflammation in milk were not significantly (P greater than 0.05) different for flunixin meglumine vs saline solution controls. The clinical response observed was consistent with the antipyretic, analgesic, and anti-inflammatory properties of flunixin meglumine.     

Respiratory tract protection upon challenge of pigs vaccinated with attenuated porcine reproductive and respiratory syndrome virus vaccines

In this study, the efficacy of two attenuated porcine reproductive and respiratory syndrome virus (PRRSV) vaccines was assessed. The virological protection in the lungs of vaccinated pigs upon challenge was studied. Also, challenged pigs were exposed to lipopolysaccharide (LPS) to evaluate clinical protection. Six-week-old pigs were immunized intramuscularly with commercial vaccines based on either an attenuated American or an attenuated European virus strain. Non-immunized pigs and pigs intramuscularly inoculated with the virulent Lelystad strain were included as controls. Six weeks after immunization, pigs were challenged either intratracheally or intranasally with the Lelystad strain, and 3 and 6 days later intratracheally exposed to Escherichia coli LPS. After LPS administration, pigs were monitored for clinical signs. At 4 and 7 days after challenge, pigs were euthanized to determine virus quantities in broncho-alveolar lavage (BAL) fluids and in lungs. Challenge virus was recovered from three out of eight pigs that had been primo-inoculated with the Lelystad strain with titers ranging between 0.3 and 3.1 log(10). Fifteen out of sixteen pigs vaccinated with the attenuated American strain were positive for challenge virus and their mean virus titers were similar to those of non-immunized challenge controls. Eleven out of 16 pigs vaccinated with the attenuated European strain were positive for challenge virus and their mean virus titers were 2.0-2.5 log(10) lower than those of non-immunized challenge controls. Thus, the virological protection in the lungs of vaccinated pigs upon challenge was incomplete, but was more pronounced in the homologous situation. Clinical signs upon LPS exposure in both vaccinated groups were not reproducible in two experiments.     

Vaccination of newborn pigs with an attenuated strain of transmissible gastroenteritis virus.

Clinical signs of transmissible gastroenteritis were not observed in newborn pigs orally inoculated with the high-passaged vaccinal transmissible gastroenteritis virus (TO-163 strain). Vaccinal viral multiplication in digestive tract of newborn pigs fed colostrum before inoculation and kept at 21 to 22 C was diminished, but was not diminished in those fed colostrum and kept at 10 to 11 C. Other groups of newborn pigs inoculated with the attenuated vaccinal virus and kept at 18 to 22 C or at 31 to 34 C were challenge exposed with virulent intestinal virus on the 1st, 2nd, . . ., or 6th postinoculation (PI) days. In the groups kept at 18 to 22 C, 2 of 7 inoculated pigs challenge exposed with virulent virus on the 3rd PI day, 4 of 7 pigs exposed on the 4th PI day, and all of the pigs exposed on and after the 5th PI day survived the exposure. In the groups kept at 18 to 22 C, the attenuated vaccinal virus was distributed mainly in the respiratory organs and lymphatic tissues. On the contrary, in the groups kept at 31 to 34 C, all of the pigs died in 2 to 5 days after challenge exposure, and the attenuated vaccinal virus was scarcely detected in any of the pigs.     

Clinical and anti-inflammatory effects of treating endotoxin-challenged pigs with meloxicam

The clinical and anti-inflammatory effects of a single treatment of 0.4 mg meloxicam/kg bodyweight on pigs that had been challenged with Escherichia coli endotoxin were investigated. Significantly lower total clinical scores were recorded in pigs treated with meloxicam than in pigs treated with a placebo. Significantly higher mean serum concentrations of thromboxane B(2) were also recorded in pigs treated with a placebo for up to 24 hours after the challenge. The serum concentrations of acute phase proteins and specific antibody titres to E coli lipopolysaccharide were unaffected by the meloxicam. The meloxicam treatment was well tolerated.     

Effect of inhaled endotoxin on cardiopulmonary function and E-selectin expression in pigs

OBJECTIVE: To evaluate the effect of controlled exposure to inhaled lipopolysaccharides (LPS) on the pulmonary inflammatory response of anesthetized pigs. ANIMALS: Forty-seven 8- to 12-week-old domestic pigs. PROCEDURE: Pigs were anesthetized with pentobarbital, instrumented for measurement of cardiopulmonary function, and randomly assigned to receive saline (0.9% NaCI) solution or 0.25, 0.5, or 1.0 microg of LPS/kg/h for 2 or 6 hours via nebulization through the endotracheal tube. Cardiopulmonary variables were measured, ex vivo neutrophil superoxide production determined, and postmortem assessment for pulmonary neutrophil influx and modulation of adhesion molecule (E-selectin) expression was done. RESULTS: Mild changes in cardiopulmonary function were observed in response to inhaled LPS in the 2-and 6-hour groups. In pigs inhaling LPS (0.5 or 1.0 microg/kg/h) for 6 hours, there was significant pulmonary neutrophil influx observed postmortem. An increase in expression of E-selectin on pulmonary endothelial cells after 6 hours of LPS inhalation (0.5 microg/kg/h) was also observed. In contrast, there was no significant influx of neutrophils or expression of E-selectin in lungs from pigs inhaling LPS for 2 hours. CONCLUSIONS AND CLINICAL RELEVANCE: inhalation of LPS resulted in localized pulmonary inflammation characterized by neutrophil influx and increased expression of the endothelial cell adhesion molecule, E-selectin. It may be possible to relate our experimental findings to the clinical consequences of airborne LPS exposure in swine confinement facilities.     

Endotoxemia in neonatal calves given antiserum to a mutant Escherichia coli (J-5)

Endotoxemia was characterized in neonatal calves given a small amount of colostrum and smooth Escherichia coli endotoxin by small-dosage (0.5 microgram/kg of body weight), slow (5-hour) IV infusion to mimic natural conditions. Responses were compared among 22 calves freely allotted to groups treated with saline solution (group I), preimmunization plasma (PP, group II), or antiserum to the rough mutant of E coli O111:B4 (J-5, group III) before endotoxin was infused. Bovine J-5 antiserum was produced by immunization of 4 cattle with J-5 boiled cell bacterin. The antiserum titers of immunoglobulin (Ig) M, IgG1, and IgG2 to the J-5 boiled cells, as determined by enzyme-linked immunosorbent assay, were 240, 7,680, and 960, respectively. The PP had enzyme-linked immunosorbent assay titers to J-5 of 240, 480, and 60 of IgM, IgG1, and IgG2, respectively. Endotoxemia in the 3 groups was characterized by significant (P less than 0.05) time-related changes in rectal temperature, heart rate, respiratory rate, capillary refill time, oral mucous membranes, nose moistness, scleral injection, attitude, PCV, total plasma protein concentration, WBC count and differential, plasma glucose, and lactate concentrations. The only significant treatment effects on clinical or laboratory values were higher mean total plasma protein concentrations in groups II and III 10 to 30 hours after endotoxin infusion was started than that in group I and increasing mean most-severe attitude abnormality score in groups I, III, and II (P less than 0.05). The administration of bovine J-5 antiserum to neonatal calves resulted in significantly higher serum IgG1 and IgG2 titers to J-5 boiled cells (P less than 0.05), and cross-reactive IgG2 to the challenge endotoxin (P less than 0.01) than did treatment with PP or saline solution; however, this antiserum did not mitigate the effects of sublethal endotoxemia. There was a significant negative correlation between IgG2 to J-5 at base line and the mean attitude abnormality score at 4.5 hours after infusion was started (P less than 0.05).     

Hemolytic complement titers and complement C3 levels in endotoxin-induced mastitis

Escherichia coli lipopolysaccharide B was instilled through the lactiferous duct of cows to induce acute mastitis. Hemolytic complement (C) activity and C3 concentrations were determined in blood serum and in renninprecipitated whey before, and at certain times after, mastitis was induced. Hemolytic complement activity was detected in the whey only during the first 36 hours after endotoxin was instilled, whereas activity was not seen before and 48 or more hours after the endotoxin was given. The maximum titer as measured with the guinea pig RBC/bovine natural antibody system was 1:64. The C3 concentrations in normal whey (before installation of endotoxin), measured by radial immunodiffusion, were between 1% and 4% of the base-line blood serum values (pool from healthy cows). The whey concentration of C3 increased (to 5% to 18%) during the first 8 hours of mastitis. However, at 72 hours, the whey values were back to preinstillation concentrations in all quarters.