Microscopic analysis of the effect of azoxystrobin treatments on Mycosphaerella graminicola infection using green fluorescent protein (GFP)‐expressing transformants

Green fluorescent protein (GFP)‐expressing transformants were used to investigate the effects of strobilurin fungicide azoxystrobin on Mycosphaerella graminicola infection. Azoxystrobin treatments (125 or 250 g AI ha^?1) were applied at various stages of the infection process under controlled conditions. GFP transformants showed conserved in vitro sensitivity to azoxystrobin and pathogenicity. Azoxystrobin controlled over 90% of M graminicola infections when applied before or during penetration of the pathogen (15% of the incubation phase). Azoxystrobin also impaired the growth of intercellular hyphae in M graminicola post‐penetration infection stages when applied at up to 50% of the incubation phase. Incubating infections observed in treated leaves were viable, but their growth was impaired and they did not induce necrosis under controlled conditions. Reduction by half of azoxystrobin dosage had little or no effect on azoxystrobin efficiency in controlling M graminicola. The contribution of post‐penetration fungistatic effect to azoxystrobin curative properties toward M graminicola in a field situation is discussed. © 2001 Society of Chemical Industry     

Use of positive contrast rhinography and intranasal sinography for diagnosis of a nasofacial sinus tract in a dog

Twelve days after a dog fight, a 2-year-old sexually intact female Bulldog was evaluated because of subcutaneous emphysema of increasing severity throughout the dogs body. Thoracic radiography revealed severe pneumomediastinum from which free air had extended into the retroperitoneal space, resulting in pneumoperitoneum. Tracheoscopic examination did not reveal a discontinuity of the trachea, pharynx, or larynx. A breach between the nasal cavity and subcutaneous tissues of the nasal region was suspected. Further diagnostic investigations included positive contrast rhinography and intranasal sinography. Via an angiographic catheter inserted into the left naris, positive contrast intranasal sinography revealed a sinus tract extending between the left nasal cavity and the subcutaneous tissue of the dorsal aspect of the nasal planum. Resolution of subcutaneous emphysema, pneumomediastinum, and pneumoretroperitoneum began 1 day after surgical closure of the intranasal opening of the sinus tract. To the authors' knowledge, this radiographic technique has not been reported.     

Antegrade pyelography for suspected ureteral obstruction in cats: 11 cases (1995-2001)

OBJECTIVE: o determine sensitivity and specificity of radiography, ultrasonography, and antegrade pyelography for detection of ureteral obstructions in cats. DESIGN: Retrospective study. ANIMALS: 11 cats. PROCEDURE: Medical records of cats that had radiography, ultrasonography, and antegrade pyelography performed for suspected ureteral obstructions were examined. Ultrasound-guided pyelocentesis and fluoroscopic-assisted antegrade pyelography were performed on 18 kidneys in 11 cats. Obstructive ureteral lesions were confirmed in all cats by surgical or necropsy examination. Sensitivity and specificity of survey radiography, ultrasonography, and antegrade pyelography for identification of ureteral obstructions were calculated. Surgical or necropsy findings were used as the standard for comparison. RESULTS: All cats were azotemic. Mean +/- SD serum creatinine and BUN concentrations were 10.2 +/- 6.1 and 149 +/- 82 mg/dL, respectively. Fifteen of 18 ureters were found to be obstructed at surgery or necropsy. Sensitivity and specificity were 60 and 100% for radiography and 100 and 33% for ultrasonography, respectively, in identification of ureteral obstructions. Leakage of contrast material developed in 8 of 18 kidneys during antegrade pyelography and prevented diagnostic interpretation in 5 of 18 studies. For the 13 diagnostic studies, specificity and sensitivity were 100% by use of the antegrade pyelography technique. Correct identification of the anatomic location of the ureteral obstruction was obtained in 100% of diagnostic antegrade pyelography studies and in 60% of radiography or ultrasonography studies. CONCLUSIONS AND CLINICAL RELEVANCE: Antegrade pyelography can be a useful alternative in the diagnosis and localization of ureteral obstructions in azotemic cats, although leakage of contrast material may prevent interpretation of the study.     

Purification and renaturation of membrane neuraminidase from Haemophilus parasuis

Haemophilus parasuis, which causes polyserositis, polysynovitis, meningitis, septicemia, and pneumonia in pigs, has emerged as an increasing problem in modern swine production systems. Co-factors for and the pathogenesis of H. parasuis disease are not defined. One of the potential virulence factors of H. parasuis is its neuraminidase (sialidase). While purifying the H. parasuis neuraminidase from the membrane fraction, we developed a protocol to renature enzymatic activity after enzyme preparations were resolved electrophorectically in denaturing polyacrylamide gels. The H. parasuis neuraminidase co-resolved with recombinant neuraminidase of Vibrio cholera; thus its apparent molecular mass is 82 kilodalton (kDa). The H. parasuis neuraminidase was associated with the membrane fraction and the purification protocol removed over 99% of the H. parasuis cell protein while retaining over 90% of the neuraminidase activity. Purified protein will provide another avenue to clone the neuraminidase gene that has been refractory to cloning and the protocol will be a means to purify recombinant protein.     

Algal dermatitis in cichlids

Three varieties of a popular African cichlid aquarium species, Pseudotropheus zebra, from 2 tropical fish farms in east central Florida were submitted for diagnostic evaluation because of the development of multifocal green lesions. The percentage of infected fish in these populations varied from 5 to 60%. Fish were otherwise clinically normal. Microscopic examination of fresh and fixed lesions confirmed algal dermatitis, with light invasion of several internal organs in each group. A different alga was identified from each farm. Fish from farm A were infected with Chlorochytrium spp, whereas fish from farm B were infected with Scenedesmus spp. Because of the numbers of fish involved, bath treatments to remove the algae from affected fish from farm B were attempted, with different dosages of several common algaecides including copper sulfate pentahydrate, diuron, and sodium chloride. However, none of these treatments were successful, possibly because of the location of the algae under the scales and within the dermis, and also because of the sequestering effect of the granulomatous response. To our knowledge, this is the first report of algal dermatitis in ornamental cichlids, as well as the first report of Scenedesmus spp infection in any fish.     

Diagnosis of myeloid leukosis induced by a recombinant avian leukosis virus in commercial white leghorn egg laying flocks

Commercial white leghorn egg layer flocks being used to produce fertile eggs for human vaccine production exhibited dramatically low peaks in egg production, two to four times higher than normal weekly mortality, and high numbers of cull, nonlaying birds after the onset of sexual maturity. These lower production characteristics could not be associated with management-related problems. Gross lesions of cull and fresh dead birds necropsied showed approximately 60% lacked ovarian activity and had lesions of a bacterial bursitis or synovitis, whereas the other 40% had tumors of the viscera but not of the bursa of Fabricius. Histologic examination of tumor-containing tissues showed lesions typical of myelocytomatosis. The diagnosis of myeloid leukosis was confirmed by the isolation of a recombinant avian leukosis virus (ALV) containing the LTR of subgroup J and the envelope of subgroup B ALV. A positive polymerase chain reaction with primers specific for the 3' untranslated region LTR confirmed the presence of LTR of ALV-J. The source of infection with this recombinant ALV was not determined; however, it is likely that commingling of the day-old egg-type chicks with ALV-J-infected meat-type chicks in a common hatchery had contributed to this outbreak.     

Foot-and-mouth disease virus

Foot-and-mouth disease virus (FMDV) is an aphthovirus of the family Picornaviridae and the etiological agent of the economically most important animal disease. As a typical picornavirus, FMD virions are nonenveloped particles of icosahedral symmetry and its genome is a single stranded RNA of about 8500 nucleotides and of positive polarity. FMDV RNA is infectious and it replicates via a complementary, minus strand RNA. FMDV RNA replication is error-prone so that viral populations consist of mutant spectra (quasispecies) rather than a defined genomic sequence. Therefore FMDV in nature is genetically and antigenically diverse. This poses important challenges for the diagnosis, prevention and control of FMD. A deeper understanding of FMDV population complexity and evolution has suggested requirements for a new generation of anti-FMD vaccines. This is relevant to the current debate on the adequacy of non-vaccination versus vaccination policies for the control of FMD.

Résumé

Le virus de la fièvre aphteuse est un aphtovirus de la famille des Picornaviridae et l'agent de la maladie animale la plus importante sur le plan économique. En tant que picornavirus typique, le virus de la fièvre aphteuse est nu, sous forme d'icosaèdre et son génome comprend un acide ribonucléique monobrin avec environ 8500 nucléotides et une polarité positive. L'acide ribonucléique de ce virus est infectieux et il se réplique par l'intermédiaire d'un brin d'ARN moins, complémentaire. La réplication de l'acide nucléique de ce virus conduit à des erreurs, de telle sorte que les populations virales comprennent un ensemble de mutants (quasi espèce) plutôt qu'une séquence génomique bien définie. Par suite, le virus de la fièvre aphteuse est génétiquement et antigéniquement varié. Ceci entraîne des difficultés importantes pour le diagnostic, la prévention et la maîtrise de la fièvre aphteuse. Une connaissance plus approfondie de la complexité et de l'évolution de la population de ce virus a conduit à des besoins pour une nouvelle génération de vaccines aphteux. Ceci est lié au débat actuel sur le choix d'une politique de vaccination ou de non-vaccination dans la lutte contre la fièvre aphteuse.     

F4 receptor-independent priming of the systemic immune system of pigs by low oral doses of F4 fimbriae

Oral administration of F4 fimbriae of Escherichia coli induces intestinal mucosal immune responses in F4 receptor-positive (F4R(+)) pigs, but not in F4R(-) pigs. We examined whether F4 fimbriae in F4R(-) animals behave like a food antigen and can induce oral tolerance. Therefore, F4R(+) and F4R(-) pigs were fed 2mg of F4 and challenged i.m. to evaluate the effect of oral F4 on the systemic immune system. As control antigen, two different oral doses (2 and 600 mg) of OVA were used. Thirty days after the i.m. OVA challenge, the OVA-specific serum IgG titre in 600 mg-fed pigs was lower than that in non-fed animals, indicating that tolerance was induced. Conversely, in the 2mg-fed pigs a rapid increase of OVA-specific IgG with higher titres than those in non-fed pigs was seen following challenge, indicating a priming of the systemic immune system. A similar priming was seen in both F4-fed F4R(-) and F4R(+) pigs. Upon challenge, non-fed pigs displayed a primary immune response with a slow increase of F4-specific serum IgG, whereas F4-fed F4R(-) and F4R(+) pigs showed secondary responses with a rapid increase of serum IgG. This was expected in F4R(+) pigs, as in these animals oral F4 induces F4-specific antibody-secreting cells in the spleen, suggesting a priming of the systemic immune system. However, also the F4-fed F4R(-) pigs displayed a secondary response, despite the failure to detect a response upon oral F4 administration. These findings suggest that the F4 antigen, at a dose of 2 mg, behaves like a common food antigen in F4R(-) pigs, namely it induces a systemic priming.