Determination of angiotensin I-converting enzyme activity in equine blood: lack of agreement between methods of analysis

Angiotensin-I converting enzyme (ACE) is a key regulator of blood pressure, electrolytes and fluid homeostasis through conversion of angiotensin I into angiotensin II. Recently, a genetic polymorphism of the ACE gene, which accounts for 47% of the variation of ACE activity in blood, has been advocated as a biomarker of athletic aptitude. Different methods of analysis and determination of ACE activity in plasma have been used in human and equine research without a consensus of a "gold standard" method. Different methods have often been used interchangeably or cited as being comparable in the existing literature; however, the actual agreement between assays has not been investigated. Therefore, in this study, we evaluated the level of agreement between three different assays using equine plasma obtained from 29 horses. Two spectrophotometric assays using Furylacryloyl-phenylalanyl-glycyl-glycine as substrate and one fluorimetric assay utilizing o-aminobenzoic acid-FRK-(Dnp)P-OH were employed. The results revealed that the measurements from the different assays were not in agreement, indicating that the methods should not be used interchangeably for measurement of equine ACE activity. Rather, a single method of analysis should be adopted to achieve comparable results and critical appraisal of the literature is needed when attempting to compare results obtained from different assays.     

Genetic control of sorghum resistance to leaf anthracnose

A study was carried out to investigate the inheritance of resistance to anthracnose, caused by Colletotrichum sublineolum, in sorghum. Crosses between resistant and susceptible parents and backcrosses between F₁ plants and the susceptible parents were carried out under field conditions. The F₁ generations and the segregant populations were evaluated under artificial inoculation conditions in the greenhouse. In the F₁ generation of all crosses with the respective isolates, all of the plants presented a resistance reaction except for the F₁ plants resulting from the BR009 × SC283 cross. In the F₂ generation, the frequencies of resistant and susceptible plants conformed to the hypothesis that one gene with two alleles controls host resistance, except in one cross. Out of the eight backcrosses, six presented segregation that corresponded to the hypothesis formulated. For most crosses, resistance was dominant, and the proportions of resistant and susceptible plants in the segregant populations conformed to the frequencies expected under the hypothesis of gene‐for‐gene resistance and dominant gene action.     

Evaluation of hepatitis E virus infection between different production systems of pigs in Brazil

The objectives of this study were to (1) investigate the occurrence of hepatitis E virus (HEV) in pigs from large-scale and family-scale farms, (2) genetically characterize the strains isolated, and (3) study the pathogenesis of swine HEV infection via immunohistochemistry. A total of 50 pigs from 10 farms in Mato Grosso State, Brazil were divided according to type of production system into either large-scale farms (n?=?5) or family-scale farms (n?=?5). Samples of liver, gallbladder, small and large intestines, bile, and feces from the pigs were analyzed by nested PCR with primers targeting the ORF2 region of HEV and by immunohistochemistry. Of the eight HEV-positive samples from pigs of family-scale farms, phylogenetic analysis revealed that seven of the swine HEV isolates clustered with subtype 3b of genotype 3 and one isolate was categorized with subtype 3 f. The HEV antigen was detected mainly in the small intestine samples from family-scale farms, suggesting an early stage HEV infection. HEV was not detected in the samples of pigs from large-scale farms, reinforcing the need for additional studies to evaluate the risk of transmission of HEV to humans from pigs from family-scale farms in Mato Grosso State.     

Prevalence of antibodies against Toxoplasma gondii among Brazilian white-eared opossums (Didelphis albiventris)

Considering that Toxoplasma gondii is a parasite of global importance which affects several animal species including humans, the current study aimed to investigate the prevalence of antibodies against T. gondii among 72 white-eared opossums (Didelphis albiventris) from Botucatu Municipality (22°53'S 48°26'W), S?o Paulo State, Brazil. The investigation was carried out from January 2008 to December 2009, when the animals had their blood samples collected and subjected to the modified agglutination test (MAT); 12 specimens had brain samples bioassayed in mice. Seroprevalence was 5.5% (n=4) and bioassays were negative. Older animals had higher prevalence of antibodies against T. gondii. Opossums in closer contact with the urban environment are likely more exposed to T. gondii than animals from the sylvatic environment.     

Identification of B-cell epitopes on the betanodavirus capsid protein

The pepscan procedure was used to identify betanodavirus B-cell epitopes recognized by neutralizing mouse monoclonal antibodies (MAbs) and serum samples obtained from sea bass, Dicentrarchus labrax, naturally infected with betanodavirus. Pepscan was performed with a panel of thirty-four 12-mer synthetic peptides that mimicked the entire betanodavirus capsid protein. Sea bass serum samples reacted strongly with three regions of the capsid protein comprising amino acid residues 1-32, 91-162 and 181-212. The latter region was also recognized by neutralizing MAbs and coincided with a region of high antigenic propensity identified by an antigen prediction algorithm. These data suggest that a region of the betanodavirus capsid protein spanning amino acid residues 181-212 may represent a neutralization domain that could potentially be used to inform the development of nodavirus vaccines and immunodiagnostic reagents.     

Haemonchus contortus resistance to ivermectin and netobimin in Brazilian sheep.

Suffolk, Texel, Hampshire Down and Ile de France sheep from the municipalities of Porto Amazonas, Piraquara and Araucaria in the State of Paraná, and Bagé in the State of Rio Grande do Sul were brought to Sobral, State of Ceará, to be used in a cross-breeding project. On arrival they had clinical signs of nematode parasitosis, and one Suffolk female died. The animals were treated orally with ivermectin (0.2 mg kg-1) and fifteen days later with netobimin (20.0 mg kg-1). Neither drug reduced the egg counts (measured in eggs per gram, EPG) significantly, and this suggested that the nematodes in the sheep were resistant to the anthelmintics used. Haemonchus contortus was the species involved. The egg counts were reduced after oral treatment with trichlorfon (100.0 mg kg-1). Haemonchus contortus larvae obtained from these animals before trichlorfon treatment and passaged through two nematode-free sheep were used in a further experiment. Twenty 6- to 9-month-old nematode-free lambs were infected with the H. contortus larvae (10,000 per animal) and after the infection was confirmed, were randomly divided into four groups of five animals. Group I was orally treated with ivermectin at 0.2 mg kg-1, Group II with oral netobimin at 20.0 mg kg-1, Group III with oral trichlorfon at 100.0 mg kg-1 and Group IV was a non-treated control. Egg counts and faecal cultures were taken before dosing on the day of treatment and seven days later when all animals were necropsied and the nematodes were collected from the abomasa and counted.(ABSTRACT TRUNCATED AT 250 WORDS)     

Flow cytometric assay in peripheral blood of dogs--reference values for leukocytes from Brazilian beagles

Use of domestic reference values in the flow cytometry analysis is known to improve its accuracy by integrating local variations as gender, race and age. Up to date application of flow cytometry in veterinary medicine has been limited to describe the percentual values just for peripheral lymphocytes subsets of blood. We now report establishment of reference values for a wide range of proportional and absolute numbers of peripheral blood leukocytes, including T cells subsets, B cells, monocytes and eosinophils, applicable to the healthy population of Beagles in Brazil and other regions with similar demographic characteristics. Normal reference values were also established to estimate the gender-related differences. This information will provide clinical aid in the evaluation of immunologic status as well as standard values for experimental animals of dogs from Brazil and other similar regions.