Thromboelastographic evaluation of the effectiveness of choline or CDP-choline treatment on endotoxin-induced hemostatic alterations in dogs.

Sepsis/endotoxemia associates with coagulation abnormalities. We showed previously that exogenous choline treatment reversed the changes in platelet count and function as well as prevented disseminated intravascular coagulation (DIC) in endotoxemic dogs. The aim of this follow-up study was to evaluate the effect of treatment with choline or cytidine-5'-diphosphocholine (CDP-choline), a choline donor, on endotoxin-induced hemostatic alterations using thromboelastography (TEG). Dogs were randomized to six groups and received intravenously (iv) saline, choline (20 mg/kg) or CDP-choline (70 mg/kg) in the control groups, whereas endotoxin (0.1 mg/kg, iv) was used alone or in combination with choline or CDP-choline at the same doses in the treatment groups. TEG variables including R- and K-time (clot formation), maximum amplitude (MA) and α-angle (clot stability), G value (clot elasticity), and EPL, A, and LY30 (fibrinolysis), as well as overall assessment of coagulation (coagulation index - CI), were measured before and at 0.5-48 h after the treatments. TEG parameters did not change significantly in the control groups, except for CI parameter after choline administration. Endotoxemia resulted in increased R-time and A value (P < 0.05), decreased K-time (P < 0.05), α-angle (P < 0.001) and CI values (P < 0.01) at different time points. Treatment with either choline or CDP-choline attenuated or prevented completely the alterations in TEG parameters in endotoxemic dogs with CDP-choline being more effective. These results confirm and extend the effectiveness of choline or CDP-choline in endotoxemia by further demonstrating their efficacy in attenuating or preventing the altered viscoelastic properties of blood clot measured by TEG.

Influence of Escherichia coli lipopolysaccharide-induced endotoxemia on plasma and tissue disposition of florfenicol after intramuscular administration in rabbits.

To assess the effects of the acute inflammatory response (AIR) induced by Escherichia coli lipopolysaccharide (LPS) on plasma and tissue disposition of florfenicol (FFC) and its metabolite florfenicol amine (FFC-a), after its intramuscular (IM) administration, twenty-two New Zealand rabbits were randomly distributed in two experimental groups: Group 1 (LPS) was treated with three intravenous doses of 2 µg LPS/kg bw, before an intramuscular dose of 20 mg/kg FFC twenty-four h after the first LPS or SS injection; Group 2 (Control) was treated with saline solution (SS) in equivalent volumes as LPS-treated group. Blood samples were collected before (T0) and at different times after FFC administration. Acute inflammatory response was assessed in a parallel study where significant increases in body temperature, C-reactive protein concentrations and leukopenia were observed in the group treated with LPS. In another two groups of rabbits, 4 h after FFC treatment, rabbits were euthanized and tissue samples were collected for analysis of FFC and FFC-a concentrations. Pharmacokinetic parameters of FFC that showed significantly higher values in LPS-treated rabbits compared with control rabbits were absorption half-life, area under the curve, mean residence time and clearance /F (Cl/F). Elimination half-life and mean residence time of FFC-a were significantly higher in LPS-treated rabbits, whereas the metabolite ratio of FFC-a decreased significantly. Significant differences in tissue distribution of FFC and FFC-a were observed in rabbits treated with LPS. Modifications in plasma and tissue disposition of FFC and FFC-a were attributed mainly to haemodynamic modifications induced by the AIR through LPS administration.

Lipolysis inhibition as a treatment of clinical ketosis in dairy cows: A randomized clinical trial.

Excessive and protracted lipolysis in adipose tissues of dairy cows is a major risk factor for clinical ketosis (CK). This metabolic disease is common in postpartum cows when lipolysis provides fatty acids as an energy substrate to offset negative energy balance. Lipolysis in cows can be induced by the canonical (hormonally induced) and inflammatory pathways. Current treatments for CK focus on improving glucose in blood (i.e., oral propylene glycol [PG], or i.v. dextrose). However, these therapies do not inhibit the canonical and inflammatory lipolytic pathways. Niacin (NIA) can reduce activation of the canonical pathway. Blocking inflammatory responses with cyclooxygenase inhibitors such as flunixin meglumine (FM) can inhibit inflammatory lipolytic activity. The objective of this study was to determine the effects of including NIA and FM in the standard PG treatment for postpartum CK on circulating concentrations of ketone bodies. A 4-group, parallel, individually randomized trial was conducted in multiparous Jersey cows (n = 80) from a commercial dairy in Michigan during a 7-mo period. Eligible cows had CK symptoms (lethargy, depressed appetite, and milk yield) and hyperketonemia (blood ß-hydroxybutyrate [BHB] ≥1.2 mmol/L). Cows with CK were randomly assigned to 1 of 3 groups where the first group received 310 g of oral PG once per day for 5 d; the second group received PG for 5 d + 24 g of oral NIA once per day for 3 d (PGNIA); and the third group received PG for 5 d + NIA for 3 d + 1.1 mg/kg i.v. FM once per day for 3 d (PGNIAFM). The control group consisted of cows that were clinically healthy (HC; untreated; BHB <1.2 mmol/L, n = 27) matching for parity and DIM with all 3 groups. Animals were sampled at enrollment (d 0), and d 3, 7, and 14 to evaluate ketone bodies and circulating metabolic and inflammatory biomarkers. Effects of treatment, sampling day, and their interactions were evaluated using mixed effects models. Logistic regression was used to calculate the odds ratio (OR) of returning to normoketonemia (BHB <1.2 mmol/L). Compared with HC, enrolled CK cows exhibited higher blood concentrations of dyslipidemia markers, including nonesterified fatty acids (NEFA) and BHB, and lower glucose and insulin levels. Cows with CK also had increased levels of biomarkers of pain (substance P), inflammation, including lipopolysaccharide-binding protein, haptoglobin, and serum amyloid A, and proinflammatory cytokines IL-4, MCP-1, MIP-1α, and TNFα. Importantly, 72.2% of CK cows presented endotoxemia and had higher circulating bacterial DNA compared with HC. By d 7, the percentage of cows with normoketonemia were higher in PGNIAFM = 87.5%, compared with PG = 58.33%, and PGNIA = 62.5%. At d 7 the OR for normoketonemia in PGNIAFM cows were 1.5 (95% CI, 1.03-2.17) and 1.4 (95% CI, 0.99-1.97) relative to PG and PGNIA, respectively. At d 3, 7, and 14, PGNIAFM cows presented the lowest values of BHB (PG = 1.36; PGNIA = 1.24; PGNIAFM = 0.89 ± 0.13 mmol/L), NEFA (PG = 0.58; PGNIA = 0.59; PGNIAFM = 0.45 ± 0.02 mmol/L), and acute phase proteins. Cows in PGNIAFM also presented the highest blood glucose increment across time points and insulin by d 7. These data provide evidence that bacteremia or endotoxemia, systemic inflammation, and pain may play a crucial role in CK pathogenesis. Additionally, targeting lipolysis and inflammation with NIA and FM during CK effectively reduces dyslipidemia biomarkers, improves glycemia, and improves overall clinical recovery.

Endotoxic kidney injury in Beagle dogs assessed by serum creatinine and symmetric dimethylarginine, and urinary neutrophil gelatinase-associated lipocalin and clusterin.

Sepsis of Gram negative bacterial origin results in lipopolysaccharide-induced endotoxemia. This often leads to acute kidney injury (AKI) and its recognition remains a challenge and delays treatment. As renal damage occurs before a rise in serum creatinine is detected, new early biomarkers of kidney injury need to be explored. The aim of this study was to determine changes in serum parameters of renal function and urine biomarkers of renal injury. This was a descriptive study. Endotoxemia was induced intravenously in six anaesthetized Beagles (T1). To achieve normotension, dogs received fluids (T2), followed by a continuous infusion of noradrenaline and dexmedetomidine or 0.9% NaCl (T3). Ten minutes later, the dogs received fluids (T4) and noradrenaline and dexmedetomidine or 0.9% NaCl in a crossover manner (T5). At each timepoint, blood and urine were collected for serum creatinine, urea, symmetric dimethylarginine, urine protein/creatinine (UPC) ratio, urine neutrophil-gelatinase-associated lipocalin (U-NGAL), U-NGAL/creatinine ratio, urine clusterin (U-clusterin) and U-clusterin/creatinine ratio. Data were analyzed using a mixed-effect model taking into account time and stage of veterinary AKI (VAKI). Three of six dogs had a VAKI stage ≥1; one with anuria and elevated creatinine. Serum creatinine (P < 0.001), U-NGAL/creatinine ratio (P = 0.01) and U-clusterin/creatinine ratio increased over time (P < 0.01). The UPC ratio (mean (range) 0.68 (0.35-2.3) versus 0.39 (0.15-0.71) P < 0.01) and U-NGAL (3164 pg/mL (100-147,555) versus 100 (100-14,524), P = 0.01) were higher in VAKI stage ≥1 versus stage 0, respectively. Endotoxemia induced VAKI stage ≥1 in half of the dogs. Repeated measurement of selected parameters could detect AKI early.

High-protein compound yogurt with quinoa improved clinical features and metabolism of high-fat diet-induced nonalcoholic fatty liver disease in mice.

Gut microbiota dysbiosis plays a crucial role in the occurrence and progression of nonalcoholic fatty liver disease (NAFLD), which may be influenced by nutritional supplementation. Quinoa, a type of pseudocereal, has gained prominence due to its high nutritional value and diverse applications. This study aimed to determine whether yogurt containing quinoa can ameliorate NAFLD and alleviate metabolic disorders by protecting against the divergence of gut microbiota. Our findings suggested that quinoa yogurt could significantly reduce the body weight gain and fat tissue weight of high-fat diet (HFD)-fed obese mice. In addition, quinoa yogurt significantly reduced liver steatosis and enhanced glucose homeostasis and insulin sensitivity. Additional research indicates that quinoa yogurt can reduce the levels of proinflammatory cytokines (i.e., tumor necrosis factor α, IL-1ß, and IL-6) and inhibit endotoxemia and systemic inflammation. The characteristics of the gut microbiota were then determined by analyzing 16S rRNA. In addition, we discovered that the gut microbiota was disturbed by HFD consumption. Particularly, intestinal probiotics and beneficial intestinal secretions were increased, leading to the expression of glucagon-like peptide-1 in the colon, contributing to NAFLD. Furthermore, endotoxemia and systemic inflammation in HFD-fed mice were restored to the level of control mice when they were fed yogurt and quinoa. Therefore, yogurt containing quinoa can effectively alleviate NAFLD symptoms and may exert its effects via microbiome-gut-liver axis mechanisms. According to some research, the role of the enteric-liver axis may also influence metabolic disorders to reduce the development of NAFLD.

Animal board invited review: The contribution of adipose stores to milk fat: implications on optimal nutritional strategies to increase milk fat synthesis in dairy cows.

A wide range of nutritional and non-nutritional factors influence milk fat synthesis and explain the large variation observed in dairy herds. The capacity of the animal to synthesize milk fat will largely depend on the availability of substrates for lipid synthesis, some of which originate directly from the diet, ruminal fermentation or from adipose tissue stores. The mobilization of non-esterified fatty acids from adipose tissues is important to support the energy demands of milk synthesis and will therefore have an impact on the composition of milk lipids, especially during the early lactation period. Such mobilization is tightly controlled by insulin and catecholamines, and in turn, can be affected indirectly by factors that influence these signals, namely diet composition, lactation stage, genetics, endotoxemia, and inflammation. Environmental factors, such as heat stress, also impact adipose tissue mobilization and milk fat synthesis, mainly through endotoxemia and an immune response-related increase in concentrations of plasma insulin. Indeed, as proposed in the present review, the central role of insulin in the control of lipolysis is key to improving our understanding of how nutritional and non-nutritional factors impact milk fat synthesis. This is particularly the case during early lactation, as well as in situations where mammary lipid synthesis is more dependent on adipose-derived fatty acids.

Pharmacokinetics and efficacy of orally administered acetaminophen (paracetamol) in adult horses with experimentally induced endotoxemia.

BACKGROUND: Acetaminophen has been evaluated in horses for treatment of musculoskeletal pain but not as an antipyretic. OBJECTIVES: To determine the pharmacokinetics and efficacy of acetaminophen compared to placebo and flunixin meglumine in adult horses with experimentally induced endotoxemia. ANIMALS: Eight university owned research horses with experimentally induced endotoxemia. METHODS: Randomized placebo controlled crossover study. Horses were treated with acetaminophen (30 mg/kg PO; APAP), flunixin meglumine (1.1 mg/kg, PO; FLU), and placebo (PO; PLAC) 2 hours after administration of LPS. Plasma APAP was analyzed via LC-MS/MS. Serial CBC, lactate, serum amyloid A, heart rate and rectal temperature were evaluated. Serum IL-1ß, IL-6, IL-8, IL-10, and TNF-α were evaluated by an equine-specific multiplex assay. RESULTS: Mean maximum plasma APAP concentration was 13.97 ± 2.74 µg/mL within 0.6 ± 0.3 hour after administration. At 4 and 6 hours after treatment, both APAP (P = <.001, P = .03, respectively) and FLU (P = .0045 and P < .001, respectively) had a significantly greater decrease in rectal temperature compared to placebo. FLU caused greater heart rate reduction than APAP at 4 and 6 hours (P = .004 and P = .04), and PLAC at 4 hours (P = .05) after treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: The pharmacokinetics of acetaminophen in endotoxemic horses differ from those reported by previous studies in healthy horses. Acetaminophen is an option for antipyresis in clinical cases, particularly when administration of traditional NSAIDs is contraindicated.

Oral Administration of Meloxicam and Flunixin Meglumine Have Similar Analgesic Effects After Lipopolysaccharide-Induced Inflammatory Response in Thoroughbred Horses.

Flunixin meglumine (FM), a nonselective cyclooxygenase (COX) inhibitor, is most frequently selected for the treatment of equine systemic inflammatory response syndrome (SIRS)/endotoxemia. However, FM has considerable adverse effects on gastrointestinal function. The aims of this study were to compare the effect of meloxicam (MX), a COX-2 selective inhibitor commonly used in equine clinical practice, with FM, and to investigate the potential for clinical application in horses with SIRS/endotoxemia. Fifteen horses were divided into three groups of five and orally administered MX (0.6 mg/kg), FM (1.1 mg/kg), or saline as placebo at 30 minutes after LPS challenge. Clinical parameters, including behavioral pain scores, were recorded and blood for clinical pathological data was collected at various times from 60 minutes before to 420 minutes after LPS infusion. The pain score were significantly lower in both the MX and FM groups than in the placebo group, with no significant difference between them. Body temperature was significantly lower in the MX and FM groups than in the placebo group. Heart rates and respiratory rates, hoof wall surface temperature, and leukocyte counts changed similarly between the MX and FM groups. TNF-α and cortisol were lower in the FM group than in the MX group. The results suggest that MX suppresses the inflammatory response after LPS infusion and has an analgesic effect similar to that of FM. Given the adverse effects of nonselective COX inhibitors, clinical application of MX may be beneficial in horses with SIRS/endotoxemia.

Effects of a single dose of orally and rectally administered misoprostol in an in vivo endotoxemia model in healthy adult horses.

OBJECTIVE: To describe misoprostol pharmacokinetics and anti-inflammatory efficacy when administered orally or per rectum in endotoxin-challenged horses. ANIMALS: 6 healthy geldings. PROCEDURES: A randomized 3-treatment crossover design was performed with a minimum washout period of 28 days between treatment arms. Prior to endotoxin challenge (lipopolysaccharide, 30 ng/kg IV over 30 minutes), horses received misoprostol (5 µg/kg once) per os (M-PO) or per rectum (M-PR) or water as control (CON). Clinical parameters were evaluated and blood samples obtained to measure plasma misoprostol free acid concentration, leukocyte counts, and tumor necrosis factor-α (TNFα) and interleukin 6 (IL-6) leukocyte gene expression and serum concentrations. RESULTS: In the M-PO treatment arm, maximum plasma concentration and area under the concentration-versus-time curve (mean ± SD) were higher (5,209 ± 3,487 pg/mL and 17,998,254 ± 13,194,420 h·pg/mL, respectively) and median (interquartile range) time to maximum concentration (25 min [18 to 34 min]) was longer than in the M-PR treatment arm (854 ± 855 pg/mL; 644,960 ± 558,866 h·pg/mL; 3 min [3 to 3.5 min]). Significant differences in clinical parameters, leukocyte counts, and TNFα or IL-6 gene expression or serum protein concentration were not detected. Downregulation of relative gene expression was appreciated for individual horses in the M-PO and M-PR treatment arms at select time points. CLINICAL RELEVANCE: Considerable variability in measured parameters was detected among horses within and between treatment arms. Misoprostol absorption and systemic exposure after PO administration differed from previous reports in horses not administered LPS. Investigation of multidose administration of misoprostol is warranted to better evaluate efficacy as an anti-inflammatory therapeutic.

Effects of intravenous administration of peripheral blood-derived mesenchymal stromal cells after infusion of lipopolysaccharide in horses.

BACKGROUND: A systemic and dysregulated immune response to infection contributes to morbidity and mortality associated with sepsis. Peripheral blood-derived mesenchymal stromal cells (PB-MSC) mitigate inflammation in animal models of sepsis. Allogeneic PB-MSC administered IV to horses is well-tolerated but therapeutic benefits are unknown. HYPOTHESIS: After IV lipopolysaccharide (LPS) infusion, horses treated with PB-MSC would have less severe clinical signs, clinicopathological abnormalities, inflammatory cytokine gene expression, and oxidative stress compared to controls administered a placebo. ANIMALS: Sixteen horses were included in this study. METHODS: A randomized placebo-controlled experimental trial was performed. Sixteen healthy horses were assigned to 1 of 2 treatment groups (1 × 109 PB-MSC or saline placebo). Treatments were administered 30 minutes after completion of LPS infusion of approximately 30 ng/kg. Clinical signs, clinicopathological variables, inflammatory cytokine gene expression, and oxidative stress markers were assessed at various time points over a 24-hour period. RESULTS: A predictable response to IV LPS infusion was observed in all horses. At the dose administered, there was no significant effect of PB-MSC on clinical signs, clinicopathological variables, or inflammatory cytokine gene expression at any time point. Antioxidant potential was not different between treatment groups, but intracellular ROS increased over time in the placebo group. Other variables that changed over time were likely due to effects of IV LPS infusion. CONCLUSIONS AND CLINICAL IMPORTANCE: Administration of allogeneic PB-MSC did not cause clinically detectable adverse effects in healthy horses. The dose of PB-MSC used here is unlikely to exert a beneficial effect in endotoxemic horses.

Effects of pre-treatment with inulin on the kidney in lipopolysaccharide-induced endotoxemia in rats.

This study was aimed to evaluate the effects of inulin used as prebiotic on the kidney in lipopolysaccharide (LPS)-induced endotoxemia model. Wistar Albino rats were divided into four groups: Control group, LPS (endotoxemia) group, Inulin + LPS group in which LPS (1.5 mg/kg, E. coli, Serotype 0111: B4) was treated after inulin (500 mg/kg) given by gavage for 21 days and Inulin group. The animals were sacrificed 24 h after the last LPS injection. Kidney samples were taken for biochemical and immunohistochemical analyses. Total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), malondialdehyde (MDA) and myeloperoxidase (MPO) values were determined. In addition, kidney sections were stained for inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α and interleukine-6 (IL-6) expression, and leukocyte infiltration. LPS caused oxidative stress and inflammation. Inulin administration could prevent oxidative stress and lipid peroxidation. Moreover, inulin decreased iNOS, TNF-α and IL-6 expression. However, it did not change the distribution of leukocytes in kidney tissues. These results suggest to promising benefits of inulin as prebiotic in reducing the effects of endotoxemia. Further studies should be conducted to evaluate the capacity of prebiotics in endotoxemia.

Un-fractionated heparin counteracts the systemic inflammatory responses and multiple organ damages caused by endotoxaemia in sheep.

BACKGROUND: Endotoxaemia is believed to be a major cause of mortality and there are several therapeutic regimens for the treatment of this situation. OBJECTIVES: The present experimental study was conducted to evaluate acute phase response, cardiovascular and hepatorenal damages following the treatment of Ovine experimental endotoxaemia model employing unfractionated heparin (UFH). MATERIALS AND METHODS: Twenty clinically healthy 1-year-old fat-tailed ewes were randomly divided into four equal groups, comprising UFH 200, UFH 400, Ctrl+ and Ctrl-. Lipopolysaccharide (LPS) from Escherichia coli serotype O55:B5 at 0.4 µg/kg was administered intravenously to the ewes. UFH (at 200 and 400 IU/kg) was administrated to the UFH 200 and UFH 400 groups, respectively. All the ewes were evaluated clinically before and 1.5, 3, 4.5, 6 and 24 hours after LPS injection. Blood samplings were also performed at those hours. We measured serum concentrations of haptoglobin, interferon-gamma, total antioxidant status, malondialdehyde, cardiac lactate dehydrogenase, cardiac troponin-I, total bilirubin, alanine transaminase and creatinine. Serum concentrations of acute phase response, cardiovascular, hepatic and renal biomarkers and clinical parameters increased significantly following the induction of endotoxaemia in the groups receiving LPS. RESULTS: The significantly lowest concentrations of these parameters at hours 4.5 and 6 among the treatment groups belonged to the UFH 400 sheep. CONCLUSIONS: UFH could act as an anti-inflammatory mediator by decreasing inflammatory cytokines and acute phase proteins, modulating oxidative stress biomarkers and reducing multiple organ dysfunction following endotoxaemia in a dose-dependent manner. Furthermore, the anti-inflammatory effects of UFH at 400 IU/kg were significantly higher than another dose. This research examined the effect of two doses of UFH and higher doses may have more anti-inflammatory effects that require further studies.

Low molecular weight heparin reduces acute phase response and multiple organ dysfunction following Ovine experimental endotoxemia model.

Endotoxemia is one of the most common inflammatory situations leading to death of ruminants. Owing to the importance of this condition, several therapeutic regimens have been proposed, evaluated and implemented to treat endotoxemia. It has recently been suggested that low molecular weight heparin may be effective in treating endotoxemia. Thus, the present experimental study was conducted to evaluate the acute phase response and multiple organ dysfunction following the treatment of the Ovine experimental endotoxemia model employing this compound. In this regard, 20 clinically healthy 1-year old Iranian fat-tailed ewes were randomly divided into 4 equal groups, comprising LMWH 50, LMWH 100, Ctrl+, and Ctrl-. Lipopolysaccharide from Escherichia coli serotype O55:B5 at 0.4 µg/kg was intravenously administered to the ewes. Low molecular weight heparin (at 50 and 100 IU/kg) was administrated to LMWH 50 and LMWH 100 groups, respectively. Positive control (Ctrl+) received lipopolysaccharide and treated only by intravenous fluid without any drugs, and negative control (Ctrl-) only received intravenous fluids without lipopolysaccharide or any drugs. All the ewes were clinically evaluated before and 1.5, 3, 4.5, 6 and 24 h after lipopolysaccharide injection, and blood samplings were also performed at those hours. Serum concentrations of serum amyloid A, tumor necrosis factor-alpha, superoxide dismutase, glutathione peroxidase, creatine kinase-MB, homocysteine, total bilirubin, aspartate aminotransferase, and creatinine were measured. Serum concentrations of acute phase proteins, inflammatory cytokines, oxidative stress, cardiovascular, hepatic and renal biomarkers, and clinical parameters were significantly increased following the induction of endotoxemia in the groups receiving lipopolysaccharide. Significantly lower concentration of these markers was observed at 4.5 and 6 h after lipopolysaccharide administration in the sheep treated with LMWH compared to the Ctrl + group. In conclusion, low molecular weight heparin could act as an anti-inflammatory drug by decreasing cytokines and acute phase proteins, modulating oxidative stress biomarkers, and by reducing multiple organ dysfunction following the induction of endotoxemia by Escherichia coli serotype O55:B5 in Iranian fat-tailed sheep in a dose-dependent manner. Furthermore, the anti-inflammatory effects of low molecular weight heparin at 100 IU/kg were significantly higher than 50 IU/kg in the treatment of endotoxemic sheep.

Influence of Escherichia coli endotoxemia on danofloxacin pharmacokinetics in broilers following single oral administration.

As a fluoroquinolone antimicrobial agent, danofloxacin is mainly used to treat avian bacterial and mycoplasma infections. The pharmacokinetic characteristics of danofloxacin are usually explored in healthy animals, while those in endotoxemic broilers are still rare. This study aimed to investigate the pharmacokinetics of danofloxacin in endotoxemic broilers induced by Escherichia coli (E. coli) lipopolysaccharide (LPS) after single oral administration. Ten healthy 5-week-old Arbor Acres (AA) broilers with similar body weight (BW) were randomly and equally divided into LPS and control groups. The LPS group was intravenously injected with an LPS of E. coli O55: B5 at 2.5 mg/kg BW, and the control group was intravenously injected with the same volume of sterile saline. Danofloxacin was administered orally at a dose of 5 mg/kg BW immediately 1 h after the intravenous injection of LPS or sterile saline. Rectal temperature was measured at predetermined times points in all broilers, and plasma and serum samples were taken. The interleukin-6 (IL-6) levels in serum samples were detected by the enzyme-linked immunosorbent assay (ELISA) kits, and danofloxacin concentrations in plasma were detected through the high-performance liquid chromatography (HPLC) method and subjected to a compartmental analysis using Phoenix software. The LPS challenge led to biphasic adaptive changes in broiler body temperature and increased the levels of IL-6. Compared with the control group, LPS treatment significantly prolonged the time to the peak concentration (LPS: 8.75 ± 3.88 h; Control: 3.20 ± 2.20 h). However, there were no significant differences in the other pharmacokinetic parameters between both groups.

THREE CASES OF ACUTE BACTERIAL SEPSIS IN PYGMY HIPPOPOTAMUS (CHOEROPSIS LIBERIENSIS) CALVE SIBLINGS.

A multiparous pygmy hippopotamus (Choeropsis liberiensis) dam produced three consecutive calves that died acutely at 13-15 wk of age from bacterial sepsis, for which diagnostic and therapeutic intervention was not possible. Streptococcus iniae (Cases 1 and 3), Escherichia coli (Case 2), and an unidentified member of the family Pasteurellaceae (Case 1) were identified in postmortem tissues through bacterial culture followed by standard and molecular identification methods. After the loss of two calves, a series of vaccinations were administered to the dam during the third pregnancy to enhance transplacental and colostral transfer of antibodies to the calf. The third calf did not survive, and the source of the bacterial infection in these three calves was undetermined. Prior to and after the birth of the fourth calf, nutritional and nutraceutical supplements were provided to the dam and calf. Additionally, pest control around the barn was enhanced. The fourth calf survived. Pygmy hippopotamus calves at the age of 13-15 wk may have increased susceptibility to bacterial infection, possibly due to waning maternally derived immunity. The findings in these cases, combined with a previous association of S. iniae in pygmy hippopotamus deaths, suggest that this bacterium is an especially important pathogen of the endangered pygmy hippopotamus.

Lipopolysaccharide (LPS) induced inflammatory changes to differentially expressed miRNAs of the host inflammatory response.

In veterinary medicine, inflammation in swine is evaluated principally by clinical signs. This method is often unreliable when assessing large animal populations because of inconsistent interpretations of clinical observations. This study examined whether changes in miRNA expression can predict the severity of the inflammatory response in swine after administration of Escherichia coli lipopolysaccharide (LPS). Whole blood from swine challenged with LPS at 0.125 µg/kg to 2.0 µg/kg body weight was collected at 0, 1, 3, and 8 h post LPS-challenge. Mature miRNAs were extracted from plasma and quantitative real-time-PCR (qRT-PCR) was used to evaluate the 84 most abundant swine miRNAs found in plasma. The miRNA changes in expression were assessed using the comparative CT Method (ΔΔCT method) for normalization with an exogenous control. The results revealed that expression of ssc-let-7e-5p, ssc-mir-22-3p, and ssc-miR-146a-5p were the most significantly changed miRNA over the time course. At 1 h post-LPS, ssc-let-7e-5p decreased as the LPS dosage levels increased from 0.125 to 1.0 µg/kg. Similarly, as the LPS doses increased from 0.125 to 0.5 µg/kg, ssc-miR-22-3p levels significantly decreased at 1 h post-LPS. In the 2.0 µg/kg LPS, ssc-miR-146a-5p levels increased between 0 and 3 h post-LPS; however, expression was downregulated with a 145 % decrease from 3 to 8 h. The three miRNA biomarkers suggest potentially useful surrogate endpoints for the evaluation of inflammatory and endotoxemia responses in swine.

Nasal secretory protein changes following intravenous choline administration in calves with experimentally induced endotoxaemia.

Nasal secretory fluid proteomes (NSPs) can provide valuable information about the physiopathology and prognosis of respiratory tract diseases. This study aimed to determine changes in NSP by using proteomics in calves treated with lipopolysaccharide (LPS) or LPS + choline. Healthy calves (n = 10) were treated with LPS (2 µg/kg/iv). Five minutes after LPS injection, the calves received a second iv injection with saline (n = 5, LPS + saline group) or saline containing 1 mg/kg choline (n = 5, LPS + choline group). Nasal secretions were collected before (baseline), at 1 h and 24 h after the treatments and analysed using label-free liquid chromatography-tandem mass spectrometry (LCMS/MS). Differentially expressed proteins (>1.2-fold-change) were identified at the different time points in each group. A total of 52 proteins were up- and 46 were downregulated at 1 h and 24 h in the LPS + saline group. The upregulated proteins that showed the highest changes after LPS administration were small ubiquitin-related modifier-3 (SUMO3) and glutathione peroxidase-1 (GPX1), whereas the most downregulated protein was E3 ubiquitin-protein ligase (TRIM17). Treatment with choline reduced the number of upregulated (32 proteins) and downregulated proteins (33 proteins) in the NSPs induced by LPS. It can be concluded that the proteome composition of nasal fluid in calves changes after LPS, reflecting different pathways, such as the activation of the immunological response, oxidative stress, ubiquitin pathway, and SUMOylation. Choline treatment alters the NSP response to LPS.

Reference intervals of acute phase proteins in healthy Andalusian donkeys and response to experimentally induced endotoxemia.

BACKGROUND: Assessment of acute phase proteins (APPs) may allow prompt detection of diseases in donkeys, that otherwise may be missed because of the stoic behavior of donkeys. Reference intervals (RIs) of APPs measured using immunoassays and a comparison of the response of these biomarkers to a controlled inflammatory insult are lacking in donkeys. OBJECTIVES: (a) To describe the RIs for APPs in healthy Andalusian donkeys, (b) to study the effects of sex and age on APPs, and (c) to assess the early response of APPs to experimentally induced endotoxemia. ANIMALS: Seventy-three healthy Andalusian donkeys (67 for RIs and 6 for endotoxemia). METHODS: Serum amyloid A (SAA), haptoglobin (Hp), C-reactive protein (CRP), ceruloplasmin (Cp), α1-acid glycoprotein (AGP), procalcitonin (PCT), ferritin (Ft), and fibrinogen (Fb) RIs were determined. Endotoxemia was induced and samples for APP determination were obtained at regular intervals for 4 hours. RESULTS: The RIs in Andalusian donkeys were: SAA (0.1-0.6 mg/L), Hp (75-2261 mg/L), CRP (1.3-7.0 mg/L), Cp (0-745 mg/L), AGP (0-884 mg/L), PCT (0-504 pg/mL), Ft (26.9-31.8 µg/L), and Fb (115-466 mg/dL). Concentrations of SAA were higher (P < .05) in jacks. Donkeys <5 years old had higher Cp, AGP, and PCT compared to older donkeys. Concentrations of SAA and Hp were significantly increased in endotoxemic donkeys from 2 hours postinduction. CONCLUSIONS AND CLINICAL IMPORTANCE: We illustrated the importance of using species-specific RIs for APPs in donkeys and the effect of age and sex on APP concentrations. Concentrations of SAA and Hp appear to be the most useful biomarkers in donkeys in the early stages of endotoxemia.

Effects of administration of ascorbic acid and low-dose hydrocortisone after infusion of sublethal doses of lipopolysaccharide to horses.

BACKGROUND: Sepsis is associated with ascorbic acid (AA) depletion and critical illness-related corticosteroid insufficiency (CIRCI) in humans. HYPOTHESES: Intravenous infusion of lipopolysaccharide (LPS) would (a) decrease endogneous AA concentrations, (b) induce CIRCI and (c) administration of a combination of AA and hydrocortisone (HC) would have decreased indices of inflammation compared to either drug alone. ANIMALS: Thirty-two healthy horses. METHODS: Randomized placebo-controlled experimental trial. Horses were assigned to 1 of 4 groups (saline, AA and HC, AA only, or HC only). Treatments were administered 1 hour after completion of LPS infusion. Clinical signs, clinicopathological variables, pro-inflammatory cytokine gene expression and production, and plasma AA concentrations were assessed at various time points. Serum cortisol concentrations and ACTH stimulation tests were used to detect CIRCI. RESULTS: There was no effect of drug on clinical signs or pro-inflammatory cytokine gene expression or production compared to controls at any time point. Administration of AA was associated with higher blood neutrophil counts 6 hours after LPS infusion (11.01 ± 1.02 K/µl) compared to other groups (8.99 ± 0.94 K/µL; P < .009). Adminstration of HC was associated with higher blood neutrophil counts 12 hours after LPS infusion (10.40 ± 0.75 K/µl) compared to other groups (6.88 ± 0.68 K/µl; P < .001). Serum cortisol increased from 5.11 ± 1.48 µg/dL before LPS administration to 9.59 ± 1.83 µg/dL 1 h after completion of LPS infusion (T1) without an effect of treatment (P = 0.59). CONCLUSIONS AND CLINICAL IMPORTANCE: Ascorbic acid and HC appeared to protect against LPS-induced neutrophil depletion and could be considered as adjunctive therapy in horses with endotoxemia.