Marta Lizandra do Rego Leal

Uso de bicarbonato e lactato-L para correção da acidose metabólica sistêmica em bovinos com acidose láctica ruminal aguda

The efficiency of sodium bicarbonate or l-lactate for correcting systemic metabolic acidosis (SMA) caused by rumen lactic acidosis (RLA) was evaluated using six rumen-cannulated steers in a cross-over experimental design. RLA was induced by administration of sucrose, intraruminally. Twenty hours later when the animals developed an intense SMA, the steers were randomly distributed and treated intravenously either with 5l of 15 mMol/l sodium bicarbonate or L-lactate solution, infused throughout 4h. Blood samples were colleted throughout the infusion at zero, 1, 2, 3, 4, 6 and 8h, for blood gas analysis. After 1hour, both sodium bicarbonate and L-lactate solutions increased blood pH and corrected adequately the SMA. Blood bicarbonate, TCO2 and base excess concentrations were also increased at the 2nd hour with L-lactate and at the 3rd hour with bicarbonate. No differences between treatments were observed for blood pH, bicarbonate, TCO2 and base excess concentrations. Treated steers after twenty-four hours showed an effective clinical recovery. L-lactate can adequately replace bicarbonate in the correction of SMA in steers with RLA.

Effects of parasitism on cellular immune response in sheep experimentally infected with Haemonchus contortus.

This work aimed to study the possible relationships among the magnitude of abomasal worm burden and the proliferation of globular leucocytes and mucosal mast cells in the abomasal mucosa, and the white blood cell count. Eighteen Suffolk × Greyface lambs were infected with Haemonchus contortus, and 12 were kept free of nematodes. Blood samples were collected on days 0, 30, and 57 post-infection (p.i.) for leucogram determination. At day 62, all animals were euthanized to count the total number of nematodes recovered in the abomasum and to count the number of mucosal mast cells and globular leucocytes. On day 57, higher levels of parasitism corresponded to lower leucocyte counts. The infected groups had lower lymphocyte counts throughout the experimental period. Animals with higher numbers of parasites had lower neutrophil and eosinophil counts on day 57. The lower the worm burden, the greater the number of mucosal mast cells (r=-0.85; p<0.01) and globular leucocytes (r=-0.87, p<0.01) observed. The sheep most resistant to haemochosis had greater peripheral and tissue cellular immune responses.

Intravenous hypertonic saline solution (7.5%) and oral electrolytes to treat of calves with noninfectious diarrhea and metabolic acidosis

Objective The aim of this study was to compare the efficacy of treating osmotic diarrhea and dehydration in calves with hypertonic saline solution (HSS) IV, isotonic electrolyte solution (IES) PO, and a combination of these 2 solutions (HSS + IES). Experimental Design Eighteen male calves 8–30 days of age were used to evaluate the efficacy of 3 methods of fluid therapy after induction of osmotic diarrhea and dehydration. The diarrhea and dehydration were induced by administration of saccharose, spironolactone, and hydrochlorothiazide for 48 hours. The animals were randomly divided into 3 experimental groups: Group 1: 7.2% hypertonic saline solution‐HSS (5 mL/kg IV); Group 2: oral isotonic electrolyte solution IES (60 mL/kg PO); or Group 3: HSS+IES. Clinical signs and laboratory finding observed 48 hours post‐induction (Time 0) included diarrhea, dehydration, lethargy, and metabolic acidosis. Results Calves treated with HSS + IES experienced decreases in hematocrit, total protein concentration, albumin concentration, urea nitrogen concentration, and plasma volume as well as increases in blood pH, blood bicarbonate concentration, and central venous pressure between 1 and 3 hours post‐treatment. These findings also were observed in animals treated with IES, however, at a slower rate than in the HSS + IES‐treated animals. Animals treated with HSS continued to display signs of dehydration, lethargy, and metabolic acidosis 24 hours post‐treatment. Conclusion Treatment with a combination of HSS and IES produced rapid and sustainable correction of hypovolemia and metabolic acidosis in calves with noninfections diarrhea and dehydration.

Diminazene aceturate associated with sodium selenite and vitamin E in the treatment of Trypanosoma evansi infection in rats

The aim of this study was to evaluate the utilization of a standard treatment with diminazene aceturate against the infection caused by Trypanosoma evansi, associated to sodium selenite and vitamin E. In vitro tests showed trypanocidal effect related to the treatment with diminazene aceturate and sodium selenite, but vitamin E had no harmful effect on the trypanosomes. In vivo experiments utilized a total of 72 adult outbreed females rats, separated into 9 groups (A, B, C, D, E, F, G, H and I), 8 animals each. Group A was the uninfected group; groups B to I were infected with 0.2mL of blood containing 10(6) trypanosomes. Parasitemia was estimated daily by microscopic examination of blood smears. Group B served as positive control; group C was treated with diminazene aceturate; group D with sodium selenite; group E with vitamin E; group F received an association of diminazene aceturate and sodium selenite; group G received an association of diminazene aceturate and vitamin E; group H received an association of diminazene aceturate, sodium selenite and vitamin E, and group I received an association of sodium selenite and vitamin E. Diminazene aceturate was administrated in a single dose on the 3rd day post infection (PI). Sodium selenite and vitamin E were administered at the 3rd and 23rd day PI. In vivo tests showed increase of longevity in groups treated with diminazene aceturate associated with sodium selenite (groups F and H). No difference was found between groups C and E, thus the vitamin E did not increase the efficacy of treatment against T. evansi when associated to diminazene aceturate. The curative efficacy of treatments was 37.5, 87.7, 37.7 and 75% to the groups C, F, G and H, respectively. Other treatments showed no efficacy. The sodium selenite when combined with chemotherapy may represent an alternative in the treatment of trypanosomosis.

Diphenyl diselenide and sodium selenite associated with chemotherapy in experimental toxoplasmosis: influence on oxidant/antioxidant biomarkers and cytokine modulation.

SUMMARY The aim of this study was to assess the effect of sulfamethoxazole/trimethoprim (ST) supplemented with diphenyl diselenide and sodium selenite in experimental toxoplasmosis, on oxidant/antioxidant biomarkers and cytokine levels. Eighty-four BALB/c mice were divided in seven groups: group A (negative control), and groups B to G (infected). Blood and liver samples were collected on days 4 and 20 post infection (p.i.). Levels of thiobarbituric acid (TBA) reactive substances and advanced oxidation protein products (AOPP) were assessed in liver samples. Both biomarkers were significantly increased in infected groups on day 4 p.i., while they were reduced on day 20 p.i., compared with group A. Glutathione reductase (GR) activity significantly (P<0·01) increased on day 4 p.i., in group G, compared with group A. INF-γ was significantly increased (P<0·001) in both periods, day 4 (groups B, C, F and G) and 20 p.i. (groups C, F and G). IL-10 significantly reduced (P<0·001) on day 4 p.i. in group B; however, in the same period, it was increased (P<0·001) in groups C and G, compared with group A. On day 20 p.i., IL-10 increased (P<0·001) in groups F and G. Therefore, our results highlighted that these forms of selenium, associated with the chemotherapy, were able to reduce lipid peroxidation and protein oxidation, providing a beneficial immunological balance between the production of pro- and anti-inflammatory cytokines.

In vitro activity of essential oils of free and nanostructured Melaleuca alternifolia and of terpinen-4-ol on eggs and larvae of Haemonchus contortus.

Haemonchus contortus is one of the major gastrointestinal nematodes responsible for significant economic and production losses of sheep. Diseases caused by this species lack effective anthelmintic products, and the search for new compounds to replace synthetic anthelmintics has been extensive. The present investigation assesses the in vitro activity of the essential oil of melaleuca (Melaleuca alternifolia), both free (TTO) and nanostructured (nanoTTO), and terpinen-4-ol (terp-4-ol) on eggs and larvae of H. contortus. Tests of egg hatching (EHT) and inhibition of larval migration (LMIT) were used to assess the in vitro efficacy of TTO, nanoTTO and terp-4-ol. Using EHT, at a concentration of 3.5 mg/ml, 100% inhibition occurred using TTO and terp-4-ol, with LC50 values of 0.43 and 0.63 mg/ml, and LC90 values of 1.75 mg/ml and 3.12 mg/ml, respectively. NanoTTO had lower activity, with 82.6% inhibition at the same concentration. Using LMIT, TTO and nanoTTO had a similar activity with 88.0% and 84.8% inhibition, respectively, at a concentration of 56 mg/ml. Terp-4-ol had a greater effect on larvae, with 85.7% inhibition at a concentration of 56 mg/ml and 82.4% at 3.5 mg/ml, demonstrating high activity at the lowest concentration tested. Therefore, the results indicate that all substances tested showed ovicidal and larvicidal activity against H. contortus. TTO, terp-4-ol and, mainly, nanoTTO may be targeted in in vivo studies, besides being a promising line of research into the control and treatment of veterinary important helminths.

Relationship between oxidative stress and pathological findings in abomasum of infected lambs by Haemonchus contortus

The aim of this study was to evaluate the nitric oxide (NO) levels, and oxidative and antioxidant markers of lambs experimentally and naturally infected by Haemonchus contortus, and its relation to lesions in the abomasum. For experimental study, a total of 14 healthy lambs were divided into two groups with seven animals each. Group A represented the uninfected animals (control), and Group B was formed by infected animals with 15,000 larvae of H. contortus. Blood was collected on days 15, 45, and 75 post-infection (PI) to obtain serum for biochemical analysis: lactate dehydrogenase (LDH), nitrite/nitrate (NOx), advanced oxidation protein products (AOPP), and ferric reducing ability of plasma (FRAP). Parasitological stool examination (eggs per gram of feces--EPG) was performed on days 15, 45, and 75 PI to verify the evolution of the infection. On day 15 PI EPG was negative, but on days 45 and 75 PI the EPG was positive for animals from Group B. In the three periods evaluated it was observed an increase of LDH levels in serum of lambs infected with gastrointestinal nematodes, but on the other hand NOx levels were reduced on the same periods in infected animals. The AOPP and FRAP levels did not differ between groups on days 15 and 45 PI, but increased significantly on day 75 PI in infected lambs. The same variables were studied in 10 lambs naturally infected with helminths, where more than 97% corresponded to H. contortus (hematocrit and EPG values were 18.8 ± 2.5% and 7120 ± 2940, respectively). Similar to the experimental study, the levels of NOx reduced, and the levels of LDH, FRAP, and AOPP increased in serum of this animal associated inflammatory infiltrate in the mucosa of the abomasum. Therefore, during the infection by H. contortus it was observed alterations in oxidative markers, indicators of cell lesion confirmed by histological examination of the abomasum, and consequently there were changes in antioxidant levels, with the purpose of cell protection. We also conclude that helminth infection interferes with the nitric oxide metabolism.

Experimental infection by Haemonchus contortus in lambs: influence of disease on purine levels in serum.

The aim of this study was to evaluate the purine levels of lambs experimentally infected with Haemonchus contortus. A total of 12 healthy lambs were divided into two groups, composed of 6 animals each: Group A represented the healthy animals (uninfected), while in Group B the animals were infected with 15 000 larvae of H. contortus. Blood was drawn on days 15, 45 and 75 post-infection (PI) in order to perform the purine analysis (ATP, ADP, AMP, adenosine, inosine, hypoxanthine, xanthine and uric acid) by high pressure liquid chromatography (HPLC) in serum. On day 15 PI a significant (P<0·05) increase in the levels of ATP and inosine was observed in the infected animals, unlike the levels of ADP, adenosine, xanthine and uric acid which were reduced. On day 45 PI a significant (P<0·05) increase in the ATP and xanthine levels in infected animals was observed, contrasting with reduced levels of ADP and uric acid. Finally, on day 75 PI an increase occurred in the levels of ATP, adenosine and hypoxanthine in infected lambs, concomitant with a reduction in the levels of ADP and uric acid (P<0·05). These changes in purine levels may influence the inflammatory process and the pathological events.

Estudo da capacidade alcalinizante de tampões metabolizáveis em bovinos sadios

Six healthy crossbred yearling 250kg steers were infused intravenously, during four hours, with five liters of each of the six different isotonic solutions of sodium salts containing 150 mMol/l of the bicarbonate, acetate, propionate, L-lactate, D and L-lactate and chloride in a 6 x 6 latin square experimental design. Blood was collected from the jugular vein, anaerobically, for blood gas analysis, samples were obtained at different times throughout the infusion (zero, 1st, 4th and 8th h ). Urine was collect for determining the pH at the same schedule. Higher blood pH was found in cattle infused with all buffers than the chloride infused animals; higher level of blood bicarbonate and base excess were obtained at 4th and 8th h with bicarbonate and L-lactate, as well as with pCO2 at the 8th h. These results show that treatments with B and L lead to a slight accumulation of buffers in the blood, and generate a discrete metabolic alkalosis compensated organically by a small retention of CO2. Best alkalinizing effects are obtained by bicarbonate and L-lactato infusions. Bicarbonate infusions causes a high urinary pH.

Delta-aminolevulinate dehydratase activity in red blood cells of rats infected with Trypanosoma evansi

The aim of this study was to evaluate the activity of delta-aminolevulinate dehydratase (δ-ALA-D) in red blood cells of rats infected with Trypanosoma evansi and establish its association with haematocrit, serum levels of iron and zinc and lipid peroxidation. Thirty-six male rats (Wistar) were divided into 2 groups with 18 animals each. Group A was non-infected while Group B was intraperitoneally infected, receiving 7·5×106 trypomastigotes per animal. Each group was divided into 3 subgroups of 6 rats and blood was collected during different periods post-infection (p.i.) as follows: day 5 (A1 and B1), day 15 (A2 and B2) and day 30 PI (A3 and B3). Blood samples were collected by cardiac puncture to estimate red blood cell parameters (RBC), δ-ALA-D activity and serum levels of iron, zinc and thiobarbituric acid reactive substances (TBARS). Rats in group B showed a significant (P<0·05) reduction of RBC count, haemoglobin concentration and haematocrit at days 5 and 15 p.i. The activity of δ-ALA-D in blood was significantly (P<0·001) increased at days 15 and 30 p.i. δ-ALA-D activity in blood had a significant (P<0·05) negative correlation with haematocrit (r=-0·61) and haemoglobin (r=-0·70) at day 15 p.i. There was a significant (P<0·05) decrease in serum iron and zinc levels and an increase in TBARS levels (P<0·05) during infection. The δ-ALA-D activity in blood was negatively correlated with the levels of iron (r=-0·68) and zinc (r=-0·57) on day 30 p.i. It was concluded that the increased activity of δ-ALA-D in blood might have occurred in response to the anaemia in remission as heme synthesis was enhanced.