Virulence factors of avian pathogenic Escherichia coli isolated from broilers from the south of Brazil

Sixty-three Escherichia coli strains isolated from broilers with respiratory problems were examined for virulence factors, hemolysin synthesis ability, motility, hemagglutination capacity, operon pap presence, colicin production, and serum resistance. The capacity to hemagglutinate guinea pig erythrocytes was found in 53 (84.1%) of the samples, but only 30 (47.6%) agglutinated chicken erythrocytes. D-mannose-sensitive hemagglutination against guinea pig erythrocytes was found in 19 (30.2%) samples and against chicken erythrocytes, in 15 (23.8%) samples, whereas the D-mannose-resistant hemagglutination with guinea pig erythrocytes was found in 34 (54%) samples, and 13 of these (20.6%) showed this characteristic against chicken erythrocytes. Operon pap, P fimbria codifier, was detected in 26 samples in a total of 34 D-mannose-resistant samples. Colicin production was observed in 55 (87.3%) of the strains, and 41.8% presented V colicin production. Of the samples analyzed, 56 (88.9%) presented serum resistance, six (9.5%) were intermediate, and only one (1.6%) was sensitive to the action of the complement. The diversity of virulence profiles detected in the samples in this study explains in part the multifactorial characteristics of avian colibacillosis.     

Use of intraluminal nitinol stents in the treatment of tracheal collapse in a dog

Tracheal collapse is a common problem that is typically observed in middle-aged and older small-breed dogs. It is a structural, obstructive airway disease with a dynamic component that can affect the intra- and extrathoracic portions of the trachea and mainstem bronchi. Many methods of treatment have been suggested, including medical management and provision of extraluminal and intraluminal support. All techniques used to treat intrathoracic and mainstem bronchial collapse have been associated with major complications or limitations. This report describes the implantation of intraluminal nitinol stents to successfully treat intrathoracic as well as extrathoracic tracheal collapse in a dog. The stents are composed of material that has characteristics similar to those of the trachea; nitinol stents may provide a method of supporting intrathoracic tracheal and mainstem bronchial collapse in dogs.     

Oestrogen-induced bone marrow aplasia in a dog with a Sertoli cell tumour

A 10-year-old male Drahthaar dog with unilateral cryptorchidism was examined because of feminisation and myelotoxicity. Radiography and ultrasonography revealed an abdominal mass which was surgically removed. The mass was identified as a Sertoli cell tumour on histological examination. Findings on bone marrow examination were compatible with aplasia due to the oestrogens secreted by the tumoral cells. Treatment with fluids, antibiotics, whole blood transfusions, corticosteroids and lithium carbonate was unsuccessful. Survival time was 22 days after surgery.     

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.     

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.     

Evaluation of three enzyme-linked immunosorbent assays (ELISAs) for the detection of serum antibodies in sheep infected with Echinococcus granulosus

The aim of this study was to develop an immunological method for the identification of sheep infected with Echinococcus granulosus which would allow the monitoring of animals imported into countries free from hydatidosis and as an aid to countries where control schemes for the disease are in operation. Three enzyme-linked immunosorbent assays (ELISAs) were developed and validated, using as antigen either a purified 8 kDa hydatid cyst fluid protein (8kDaELISA), a recombinant EG95 oncosphere protein (OncELISA) or a crude protoscolex preparation (ProtELISA). Sera used for the assay validations were obtained from 249 sheep infected either naturally or experimentally with E. granulosus and from 1012 non-infected sheep. The highest diagnostic sensitivity was obtained using the ProtELISA at 62.7 and 51.4%, depending on the cut-off. Assay sensitivities were lower for the 8kDaELISA and the OncELISA. Diagnostic specificities were high, ranging from 95.8 to 99.5%, depending on the ELISA type and cut-off level chosen. A few sera from 39 sheep infected with T. hydatigena and from 19 sheep infected with T. ovis were recorded as positive. Western immunoblot analysis revealed that the dominant antigenic components in the crude protoscolex antigen preparation were macromolecules of about 70-150 kDa, most likely representing polysaccharides. This study demonstrated that the ProtELISA was the most effective immunological method of those assessed for detection of infection with E. granulosus in sheep. Because of its limited diagnostic sensitivity of about 50-60%, it should be useful for the detection of the presence of infected sheep on a flock basis and cannot be used for reliable identification of individual animals infected with E. granulosus.     

Use of spatial statistics and monitoring data to identify clustering of bovine tuberculosis in Argentina

The spatial distribution of endemic bovine tuberculosis (TB) in Argentine cattle herds was described using recorded information on the detection of TB-like lesions in cattle slaughtered between March 1995 and 1997 at 126 slaughterhouses with federal inspection. Approximately, 47% (9472396 cattle) of the estimated total number of cattle slaughtered in Argentina during this period was included in the study. Information on the number of cattle per source herd consigned to slaughter, number of cattle with TB-like lesions per herd and the geographical location of counties from which cattle originated was used to investigate spatial clustering of TB. Overall, no evidence of clustering of TB prevalence by county was detected (Moran's autocorrelation statistic I=0.009, P=0.089). However, first- (Cuzick and Edwards' test statistic, T(k)=87, P=0.036) and second-order (T(k)=170, P=0.038) nearest-neighbor case-counties (TB prevalence>median prevalence of all counties, 6.7%) were clustered. Using the spatial scan test based on a Bernoulli model, the most-likely cluster (P=0.001) identified during the study period included 5793 cases of TB (5.2 per 1000 km(2)) in five counties. This cluster coincided with Santa Fe Province, which contains 21% of all dairy cows in Argentina and accounts for 34% of the country's milk production. Several secondary clusters of TB-also located in dairy districts-were identified. Study results demonstrate that bovine TB is clustered in Argentina, and these clusters coincide with dairy cattle production. Identification of clustering can assist efforts to eradicate bovine TB from Argentina. Further spatial investigations need to focus on the reasons why TB is clustered in Argentina. In particular, the relationship between TB clustering and management practices-such as grazing density and production systems-need to be described to assist in the development of disease-control programs. The use of spatial statistics and geographical information systems could meet these needs.     

Brucellosis in Argentina

Brucellosis has been recognized in Argentina since the 19th century. Several studies demonstrated the presence of the disease in most of the domestic species. Actually, the estimate of prevalence is that between 10 and 13% of the farm animals are infected with bovine brucellosis with an individual rate of 4–5%. The annual economical losses have been estimated at US$ 60,000,000. The control of bovine brucellosis began in 1932 and successive resolutions have been issued since then. The current resolution indicates that B. abortus S19 is mandatory in female calves between 3 and 8 months of age. The vaccine strain B. abortus RB51 was provisionally approved but only for cattle older than 10 months of age. The brucellosis control program consists principally of test and slaughter. This methodology has been successful mainly in the dairy farms that have the incentive due to increased pricing because of obtaining a low prevalence of the disease. Brucellosis has been found in porcine, caprine, ovine and canine species. All Brucella species have been found in the country. Human brucellosis is an important disease and a national coordinated diagnostic net has been formed to better control the disease in man.     

Field and laboratory selection of Frankliniella occidentalis (Pergande) for resistance to insecticides

Response of western flower thrips, Frankliniella occidentalis (Pergande), to selection for resistance to insecticides commonly used to control this pest in Murcia (south-east Spain) was studied under field and laboratory conditions. In the field, plots within sweet pepper crops in commercial and experimental greenhouses were treated under different selection strategies: insecticide rotation versus formetanate reiteration, formetanate reiteration versus acrinathrin reiteration, and formetanate reiteration versus methiocarb reiteration. Thrips populations were sampled monthly and bioassayed against methiocarb, methamidophos, acrinathrin, endosulfan, deltamethrin and formetanate. In the laboratory, F occidentalis strains were selected against each insecticide for several generations. To evaluate cross-resistance, each selected strain was bioassayed with the other insecticides. Frankliniella occidentalis populations showed a rapid development of acrinathrin resistance, reaching high levels in field and laboratory conditions. Formetanate and methiocarb resistance were also observed, although development was slower and at moderate levels. Cross-resistances between acrinathrin/deltamethrin and acrinathrin/formetanate were detected under field and laboratory conditions. Formetanate/methiocarb cross-resistance was suspected in laboratory selections, but not in field assays. Simultaneous moderate resistance levels to the three specific insecticides against thrips (formetanate, methiocarb and acrinathrin) were shown in laboratory selection strains, indicating a general mechanism of resistance, probably metabolic.     

Assessment of relevant factors and relationships concerning human dermal exposure to pesticides in greenhouse applications

Principal component analysis (PCA) was applied to the gas chromatographic data obtained from 23 different greenhouse trials. This was used to establish which factors, including application technique (very small, small, medium and large drop-size), crop characteristics (short/tall, thin/dense) and pattern application of the operator (walking towards or away from the treated area) are relevant to the dermal exposure levels of greenhouse applicators. The results showed that the highest exposure by pesticides during field applications in greenhouses, in the climatic conditions and in the crop conditions typical of a southern European country, occurs on the lower legs and front thighs of the applicators. Similar results were obtained by hierarchical cluster analysis (HCA). Drop-size seems to be very important in determining total exposure, while height and density of crops have little influence on total exposure under the conditions of the present study. No pesticide type is a major factor in total exposure. The application of multiple regression analysis (MRA) allowed assessment of the relationships between the pesticide exposure of the less affected parts of the body with the most affected parts.