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.

Effects of intravenous butaphosphan and cyanocobalamin to late pregnant ewes on the metabolic indices around parturition and weight gain of their lambs after birth.

BACKGROUND: Management and control of metabolic disorders in sheep around parturition is important. and various researchers have suggested different managerial solutions. Butaphosphan and cyanocobalamin are widely used for curing metabolic disorders resulting from poor nutrition, inadequate management or diseases. OBJECTIVES: It was hypothesised that butaphosphan and cyanocobalamin could improve the metabolism of ewes around parturition. METHODS: Twenty-eight clinically healthy 3-year-old pregnant Afshari ewes from 21 days before parturition were enrolled into four equal groups: control (Ctrl), B+C1, B+C2 and B+C3. The Ctrl group only received intravenous normal saline and B+C1, B+C2 and B+C3 ewes, respectively, received an intravenous combination of 10% butaphosphan and 0.005% cyanocobalamin at 2, 4 and 6 ml/ewe, on Days 19-21, 10-12 and 1-3 before parturition. Blood samples were taken from all the ewes on Days 21, 12 and 3 before lambing at parturition day and on days 3, 12 and 21 after parturition. A body condition score of all the ewes was assessed at blood sampling days, and lambs born from the ewes were weighed at birth and every 2 weeks up to 3 months. Serum concentrations of glucose, cortisol, non-esterified fatty acids, beta-hydroxy butyric acid, triglyceride, cholesterol, high-, low- and very-low-density lipoproteins, aspartate aminotransferase and alanine aminotransferase were measured. RESULTS: This drug combination decreased circulating glucose, cortisol, lipid profile and hepatic enzymes via dose-dependent manner, 6 ml of this drug compound/ewe was more potent than 4 and 2 ml/ewe. The lambs' weight from mothers receiving 6 ml of this combination was significantly higher than those of the others. CONCLUSIONS: It may be suggested that the intravenous administration of 6 ml/ewe of this combination for 3 consecutive days in three states before parturition had prophylactic effects on metabolic disorders of ewes and enhanced the lambs weight gain after birth.

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.

Evaluating heart electrical activities and cardiac arrhythmias of Holstein cows during ageing by short-term electrocardiography in comparison with 24-hour holter-monitoring.

Short-term electrocardiography is one of the most suitable tools to study the electrical activity of the heart, but the use of a tool such as a Holter-monitor with the ability to assess the long-term of the heart electrical activity, can provide more accurate information about these activities by comparing the results. It is possible to understand the superiority of each over the other and the resulting differences. Therefore, 60 female Holstein cows in 10 age groups, including 1 day, 1, 3, 6 months, 1, 2, 3, 4, 6 and 8 years were included in the study (6 heads in each age group). Electrocardiography (for 5 min) and Holter-monitoring (for 24 hr) were performed from the entire study population. The Q, R and T amplitudes in electrocardiography were significantly higher than those in Holter-monitoring. The P, R and T durations and P-R, R-R, Q-T and S-T intervals at all ages were significantly longer in the Holter-monitoring than in the electrocardiographic method. The heart rate of animals was significantly lower in the 24-hr Holter-monitoring than in the short-term electrocardiography. The trend of changes of all parameters was significant during ageing. Cardiac arrhythmias included sinus arrhythmia and sino-atrial block, which were the most common cardiac arrhythmias in the 24-hr Holter-monitoring. It appears that long-term Holter-monitoring is a more reliable method than short-term electrocardiography to assess cardiac arrhythmias. Additionally, the indicators of electrical activity of the heart (waves) in the Holter-monitoring method are significantly different from short-term electrocardiography, which is probably due to the collection of information over a long period and in non-stressful situations. Furthermore, it appears that the use of the 24-hr Holter-monitoring method is preferable to the short-term electrocardiography method to evaluate the electrical activity of the heart of cows at all ages.

Intravenous Administration of Butaphosphan and Cyanocobalamin Combination to Late-Pregnant Dairy Cows Reduces Their Insulin Resistance After Calving.

Dairy cows suffer insulin resistance following parturition and lactogenesis. Several researchers attempted to reduce insulin resistance via dietary and parenteral supplementations of different substances to promote metabolic performance of dairy cows. Due to mechanisms of actions of butaphosphan in combination with cyanocobalamin, we hypothesized that this compound may reduce insulin resistance of dairy cows following parturition; hence, the effects of the intravenous administration of butaphosphan and cyanocobalamin to prepartum dairy cows on their insulin resistance after calving were evaluated. Twenty-four multiparous Holstein dairy cows were enrolled 3 weeks prior to parturition and divided into four equal groups, including control (Ctrl) and butaphosphan and cyanocobalamin (B+C) 1, 2, and 3. Ctrl cows received 15 mL of 0.9% NaCl solution and B+C 1, 2, and 3 groups intravenously received 2, 4, and 6 mL/100 kg BW of 10% butaphosphan and 0.005% cyanocobalamin combination over three periods of 3 consecutive days, including 21-19, 12-10, and 3-1 days before calving, respectively. Intravenous glucose tolerance test was performed weekly 1, 2, and 3 weeks after parturition to evaluate the insulin resistance phenomenon. Circulating levels of glucose, insulin, non-esterified fatty acids (NEFA), and beta-hydroxybutyric acid (BHBA) were assessed 1, 2, and 3 weeks after calving. Ctrl cows were the most insulin-resistant group, and B+C1 group was the most insulin-sensitive, followed by B+C2 and B+C3 groups. The NEFA and BHBA levels in the B+C3 group were significantly lower than those in the other groups. In conclusion, intravenous administration of butaphosphan and cyanocobalamin to the late-pregnant dairy cows may reduce their insulin resistance after calving.

The comparative effects of dietary monensin and propylene glycol on insulin resistance of transition dairy cows.

During the transition period, dairy cows suffer from negative energy balance due to the upcoming insulin resistance as a major metabolic disturbance. We hypothesized that providing glucose precursors for transition dairy cows may reduce the insulin resistance. In this study, 24 multiparous Holstein dairy cows were enrolled 8 weeks prior to parturition and divided into 4 equal groups, including control (Ctrl), monensin (Mo), propylene glycol (PPG), and monensin plus propylene glycol (Mo + PPG). Cows from the Mo and PPG groups received 1 mg/kg body weight (BW) of monensin, daily. Cows from the PGG group received 150 g of propylene glycol, daily. Cows from the Mo + PPG group received 1 mg/kg BW of monensin and 150 g/head of propylene glycol daily and Ctrl cows received basal diet without any supplementations. Intravenous glucose tolerance test (ivGTT) was conducted weekly from 3 weeks before to 3 weeks after parturition to evaluate the insulin resistance phenomenon. Immediately after glucose administration, glucose and insulin increased significantly, and their alterations were significant during the study. Glucose and insulin were significantly higher in the Ctrl group than in the other groups, and their levels in different pre- and post-partum periods were significantly lower in the Mo + PPG group than in the other studied groups. The results of this study represented that the supplementary feeding with propionate precursors, such as monensin and propylene glycol, reduced the insulin resistance in dairy cows during the transition period. This effect is more explicit by propylene glycol than by monensin, and the combination of both reduces insulin resistance at higher rates. The use of these dietary supplements is likely to produce more propionates as the main precursor of glucose; therefore, it reduces the negative energy balance and subsequently decreases the insulin resistance. In this regard, to reduce insulin resistance, it is recommended that dairy cows during the transition period be fed with monensin and propylene glycol supplements.

Effects of Polymyxin B on Clinical Signs, Serum TNF-α, Haptoglobin and Plasma Lactate Concentrations in Experimental Endotoxaemia in Sheep.

INTRODUCTION: The experiment evaluated the effects of intravenous administration of polymyxin B on experimental endotoxaemia in sheep. MATERIAL AND METHODS: Twenty clinically healthy fat-tailed sheep were randomly divided into: a group treated with 6,000 U/kg of polymyxin B, a group at 12,000 U/kg, and positive and negative controls. Endotoxaemia was induced by intravenous administration of lipopolysaccharide (LPS) from E. coli serotype O55:B5 at 0.5 µg/kg. polymyxin was infused intravenously along with 2.5 L of isotonic intravenous fluids at 20 mL/kg/h. The positive control group received LPS and 2.5 L of isotonic fluids, the negatives receiving just 2.5 L of isotonic fluids. Clinical signs were evaluated before and at 1.5, 3, 4.5, 6, 24, and 48 h after LPS administration. Blood was also sampled at the denoted hours and serum haptoglobin, tumour necrosis factor-α (TNF-α), and plasma lactate concentrations were assayed. RESULTS: The serum concentration of TNF-α in the positive control group increased significantly up to 48 h after LPS administration. The concentration of TNF-α was significantly different from those of the polymyxin B and positive control groups from 3 to 48 h; also, the concentrations of haptoglobin at different times in the polymyxin groups were lower than those of the positive control group and were significant at hours 3 to 48 (P < 0.05). Following the LPS administration, haptoglobin and TNF-α concentrations changed without significant difference between the two polymyxin B groups. CONCLUSION: Polymyxin B (6,000 U/kg) restrained blood lactate concentrations. Furthermore, it significantly improved the clinical signs in endotoxaemic animals, including rectal temperature and heart and respiratory rates. Polymyxin B may be an antiendotoxic in fat-tailed sheep.

The comparative efficacy of tyloxapol versus pentoxifylline against induced acute phase response in an ovine experimental endotoxemia model.

The effective treatments of endotoxemia are necessary to prevent high mortality rates. Hence, the present study was performed to clarify the antiendotoxic effects of tyloxapol and pentoxifylline in experimentally induced endotoxemia in sheep. Thirty clinically healthy 1-year-old Iranian fat-tailed ewes were randomly divided into six equal experimental (n = 5) groups, comprising Negative and Positive control, Tyloxapol 1, Tyloxapol 2, Pentoxifylline 1 and Pentoxifylline 2. Phenol extracted lipopolysaccharide from Escherichia coli serotype O55:B5 was infused at 2 µg/kg intravenously. Tyloxapol (200 and 400 mg/kg) and pentoxifylline (30 and 60 mg/kg) were injected to Tyloxapol and Pentoxifylline groups, respectively, at 90 min after endotoxemia induction over 60 min along with intravenous fluids. Blood samples were collected from all ewes prior and 1.5, 3, 4.5, 6, 24 and 48 h after lipopolysaccharide injection and sera and plasmas were separated, subsequently. Haptoglobin, serum amyloid A, tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), superoxide dismutase and glutathione peroxidase were measured in all samples. Serum concentrations of haptoglobin, serum amyloid A, TNF-α and IFN-γ in Tyloxapol 1 and 2 and Pentoxifylline 1 and 2 groups were significantly lower than Positive control one after hour 3. There were no significant differences among Tyloxapol and Pentoxifylline groups (P > 0.05). Superoxide dismutase and glutathione peroxidase activities in Tyloxapol 1 and 2 and Pentoxifylline 1 and 2 groups were significantly lower than Positive control one after hour 3. There were no significant differences among Tyloxapol 1 and 2 and Pentoxifylline 1 and 2 groups (P > 0.05). Tyloxapol and pentoxifylline act as the anti-inflammatory mediators by decreasing pro-inflammatory cytokines and hepatic APPs and modulating oxidative enzymes activity after endotoxemia induction in sheep. Furthermore, their efficacies at different doses were significantly similar together and both drugs don't induce their effects by dose dependent manner and the anti- and pro-inflammatory effects of them were statistically similar.

Genetic characterization and phylogenetic analysis of Trypanosoma evansi in Iranian dromedary camels.

Whole blood samples were collected from 117 male clinically healthy Camelus dromedarius aged between 6 months to 18 years from several farms in Yazd Province of Iran. Trypanosoma evansi-affected camels were detected by Giemsa-stained blood smears, and the positive blood samples (4 out of 117) were submitted to PCR examination and phylogenetic analysis. Basic Local Alignment Search Tool data of the obtained complete internal transcribed spacer (ITS) sequences revealed that they corresponded to those of T. evansi, Thailand cattle isolate (AY912276) with the homology of 99 %. Both phylogenetic trees generated by ITS1 and complete ITS were unable to clearly show inter- and intraspecific genetic diversity of Trypanosoma spp. isolates. The phylogenetic tree inferred from the ITS2 nucleotide sequences (569 bp) clearly showed the genetic diversity of the parasites. Phylogenetic and molecular analyses of this region showed that two distinct genotypes of T. evansi in Iranian dromedary camels are present. In contrast to the ITS1 and ITS2 regions, multiple alignment of the nucleotide sequence of the 5.8S rRNA showed a high degree of sequence conservation during evolution in various Trypanosoma spp.

Anti-inflammatory effects of insulin regular and flunixin meglumine on endotoxemia experimentally induced by Escherichia coli serotype O55:B5 in an ovine model.

BACKGROUND: Endotoxemia is a major cause of mortality in large animals and there are several therapeutic regimens for the treatment of endotoxemia. Recent studies have suggested the anti-inflammatory effects of insulin in endotoxemic human and laboratory animal models but to the best of our knowledge there is no report on the possible therapeutic effect of insulin in large animal endotoxemia. OBJECTIVE: This experiment was conducted to evaluate the anti-inflammatory effects of insulin regular compared with flunixin meglumine on the treatment of endotoxemia in sheep. METHODS: Lipopolysaccharide from Escherichia coli was administered intravenously to ewes. Anti-inflammatory effects of flunixin meglumine (at 2.2 mg/kg) and insulin regular (at 1.5 and 3 IU/kg) were evaluated by determination of serum concentrations of acute phase proteins, inflammatory cytokines and oxidative stress biomarkers. RESULTS: Insulin regular at 3 IU/kg controlled the acute phase response following endotoxemia induction. The anti-inflammatory potency of insulin regular at 3 IU/kg was significantly higher than at 1.5 IU/kg and of flunixin meglumine at 2.2 mg/kg (P < 0.05). CONCLUSION: Insulin regular induces its anti-inflammatory effects in a dose-dependent manner. Intravenous use of insulin regular can be a potential new therapeutic regimen for endotoxemia in large animal medicine.