Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Effects of the aglycone of ascaulitoxin on amino acid metabolism in Lemna paucicostata

Ascaulitoxin and its aglycone (2,4,7-triamino-5-hydroxyoctanoic acid, CAS 212268-55-8) are potent phytotoxins produced by Ascochyta caulina, a plant pathogen being developed for biocontrol of weeds. The mode of action of this non-protein amino acid was studied on Lemna paucicostata. Ascaulitoxin is a potent growth inhibitor, with an I₅₀ for growth of less than 1 μM, almost completely inhibiting growth at about 3 μM. Its action is slow, starting with growth inhibition, followed by darker green fronds, and then chlorosis and death. Most amino acids, including non-toxic non-protein amino acids, reversed the effect of the toxin when supplemented in the same medium. Supplemental sucrose slightly increased the activity. d-Amino acids were equally good inhibitors of ascaulitoxin activity, indicating the amino acid effects may not be due to inhibition of amino acid synthesis. Oxaloacetate, the immediate precursor of aspartate, also reversed the activity. LC-MS did not detect interaction of the compound with lysine, an amino acid that strongly reversed the effect of the phytotoxin. Metabolite profiling revealed that the toxin caused distinct changes in amino acids. Reduction in alanine, paralleled by enhanced levels of the branched chain amino acids valine, leucine and isoleucine and nearly unchanged levels of pyruvate, might indicate that the conversion of pyruvate to alanine is affected by ascaulitoxin aglycone. In addition, reduced levels of glutamate/glutamine and aspartate/asparagine might suggest that synthesis and interconversion reactions of these amino group donors are affected. However, neither alanine aminotransferase nor alanine: glyoxylate aminotransferase were inhibited by the toxin in vitro. Our observations might be explained by three hypotheses: (1) the toxin inhibits one or more aminotransferases not examined, (2) ascaulitoxin aglycone affects amino acid transporters, (3) ascaulitoxin aglycone is a protoxin that is converted in vivo to an aminotransferase inhibitor.     

Characterization of lettuce (Lactuca sativa L.) grown with biopesticides and deltamethrin

The purpose of this study was to evaluate the physico-chemical characteristics and the post-harvest loss of mass of curly lettuce (Lactuca sativa L.) cv. Verônica that was grown in a protective environment with applications of biopesticides. The curly lettuce was treated by spraying with neem oil, cassava flour wastewater (manipueira), biofertiliser or the control treatment of deltamethrin. We determined the weight, circumference, dry matter, ash, crude protein, total phenols and minerals profile for the heads of lettuce. After harvesting, the lettuce was packed into opened plastic bags and kept at 5 °C for 5 days to evaluate its weight loss. No differences were observed among the treatments for the physico-chemical parameters. The lettuce that was sprayed with manipueira or biofertiliser had the greatest weight loss during storage. The achievement of similar yields with cheaper products suggests a benefit of using organic bioinsecticides.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Effect of the number of intercrosses on grain yield in basic rice synthetic populations

The effect of 0, 1, 2, 3 and 4 intercross cycles using genetic male sterility on the mean yield and genetic variability of the CNA 5 irrigated rice synthetic population was assessed studying 300 S_0:2 (1998/99) and S_0:3 (1999/00) families grown in two locations (Lambari, MG and Goianira, GO). Each intercross was investigated using a 8 × 8 triple lattice design experiment containing 60 families and four control cultivars. Five experiments were carried out per location and generation, totaling 20 experiments. Individual, combined over locations, combined over generations and combined over locations and generations analyses of variance were performed. Linear regression equations were fitted to data of each location and generation as well as, genetic and phenotypic parameters were estimated. An average increase of 3.91% in grain yield was obtained with the increase in the number of intercrosses. However, when genetic gains obtained from selection of the five and ten highest yielding families were considered, negative b estimates were always obtained, suggesting a tendency of gain increase reduction with the increase in the number of intercrosses. This was confirmed by the analysis of the means of the best families, since increases in the number of intercrosses did not result in selection of families with greater mean grain yield. The estimates of the genetic variances and heritabilities did not suggest an increase in genetic variability with the increase in intercross cycles. Therefore, carrying out intercrosses on the base population did not improve the results of the irrigated rice recurrent selection program. This revised version was published online in August 2006 with corrections to the Cover Date.     

Influence of selenium on aflatoxin B1 or crotalaria toxicity in turkey poults

This research compared the toxic effects of aflatoxin B1 and monocrotaline, the active principle of Crotalaria spectabilis, and the additive effect between aflatoxin B1 and monocrotaline in turkey poults. It was of interest whether selenium fed at dosage levels of 0.1, 5, or 10 micrograms/g of feed would protect against the toxic effect of aflatoxin and/or monocrotaline, and whether the toxicants would result in detectable residues in poult tissues. A total of 180 healthy 1-day-old male turkey poults was assigned at random to 12 treatment groups (15 birds/group). Body and liver weight losses, and low serum concentrations in total protein (TP), albumin (A), alpha-globulin (alpha G), and beta-globulin (beta G), as well as high values in gamma-globulin (gamma G), were produced in the groups fed crotalaria. Pathologic changes were induced by monocrotaline with no protection afforded by the added selenium. Low values in TP, A, alpha G, and beta G and in body and liver weights were observed in groups given the combination of aflatoxin plus crotalaria. Gross lesions were associated with an additive toxic effect and a lack of protective effect of selenium against this combination. However, higher values in TP, A, alpha G, and beta G, and liver weights in groups fed aflatoxin B1 plus selenium indicated that selenium had a protective effect against aflatoxin toxicity. Residues of aflatoxin B1 and aflatoxin M1 were found in the kidneys of poults fed aflatoxin B1; also, dehydroretronecine (the metabolite of monocrotaline) was detected in livers of poults fed Crotalaria spectabilis seeds.     

A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.     

Pre- and postoperative force plate analysis of dogs with experimentally transected cranial cruciate ligaments treated using tibial plateau leveling osteotomy

Objective— Quantitative and objective assessment of hindlimb kinetics after cranial cruciate ligament (CrCL) transection and subsequent stifle stabilization using the tibial plateau leveling osteotomy (TPLO) in normal dogs.
Study Design— In vivo experimental biomechanical evaluation.
Animals— Six healthy adult foxhounds.
Methods— Dogs were screened by orthopedic and radiographic examination before study entry. Force plate analysis of gait was measured before extirpation of the right CrCL and TPLO and again at 8 and 18 weeks after surgery.
Results— There was a significant decrease in peak vertical forces (PVFs) and vertical impulse (VI) of the treated hindlimb at 8 weeks when compared with preoperative and 18-week measurements. When compared with preoperative values, there was no significant difference in 18 week PVF and VI in dogs that had TPLO.
Conclusion— TPLO can restore kinetic measures of limb function at 18-weeks after surgery when compared with preoperative values after experimental transection of the CrCL in dogs.
Clinical Relevance— TPLO induces lameness that returns to near normal at 18 weeks. The severity and duration of lameness was similar to that reported for other experimental models of stifle instability repaired by different techniques.