Variation in Pathogenicity Among South African Isolates of Phytophthora cinnamomi

Phytophthora cinnamomi isolates from South Africa were evaluated for differences in growth rate in vitro and levels of pathogenicity towards Eucalyptus smithii in the field. Inoculations were conducted in the field in summer and winter in two subsequent years at two locations in South Africa using 59 P. cinnamomi isolates. The isolates differed significantly in growth rate in vitro, as well as in levels of pathogenicity to E. smithii in the field. Growth rate in vitro was significantly influenced by interactions with culture age, geographic origin and genetic background as determined using isozymes. Levels of pathogenicity in the field were influenced by season of inoculation and average minimum temperatures at trial sites. The host from which P. cinnamomi isolates were originally obtained did not significantly affect levels of pathogenicity in the field. Culture age had a significant negative effect on growth rate in vitro and pathogenicity in the field. Significant differences in levels of pathogenicity could be found for different multilocus isozyme genotypes. Geographic origin and mating type of P. cinnamomi isolates had no significant effect on levels of pathogenicity in the field. A positive correlation was found between growth rate in vitro and levels of pathogenicity in the field. Levels of variation for pathogenicity within A1 mating type isolates were significantly lower than for A2 mating type isolates. Results of this study provide valuable information on selection of P. cinnamomi isolates for future resistance/tolerance screening assays of Eucalyptus germplasm in South Africa.     

Investigation of the expression and localization of glucose transporter 4 and fatty acid translocase/CD36 in equine skeletal muscle

OBJECTIVE: To investigate the expression and localization of glucose transporter 4 (GLUT4) and fatty acid translocase (FAT/CD36) in equine skeletal muscle. SAMPLE POPULATION: Muscle biopsy specimens obtained from 5 healthy Dutch Warmblood horses. PROCEDURES: Percutaneous biopsy specimens were obtained from the vastus lateralis, pectoralis descendens, and triceps brachii muscles. Cryosections were stained with combinations of GLUT4 and myosin heavy chain (MHC) specific antibodies or FAT/CD36 and MHC antibodies to assess the fiber specific expression of GLUT4 and FAT/CD36 in equine skeletal muscle via indirect immunofluorescent microscopy. RESULTS: Immunofluorescent staining revealed that GLUT4 was predominantly expressed in the cytosol of fast type 2B fibers of equine skeletal muscle, although several type 1 fibers in the vastus lateralis muscle were positive for GLUT4. In all muscle fibers examined microscopically, FAT/CD36 was strongly expressed in the sarcolemma and capillaries. Type 1 muscle fibers also expressed small intracellular amounts of FAT/CD36, but no intracellular FAT/CD36 expression was detected in type 2 fibers. CONCLUSIONS AND CLINICAL RELEVANCE: In equine skeletal muscle, GLUT4 and FAT/CD36 are expressed in a fiber type selective manner.     

Immunohistochemical identification and fiber type specific localization of protein kinase C isoforms in equine skeletal muscle

OBJECTIVE: To investigate whether protein kinase C (PKC) isoforms are expressed in equine skeletal muscle and determine their distribution in various types of fibers by use of immunofluorescence microscopy. ANIMALS: 5 healthy adult Dutch Warmblood horses. PROCEDURE: In each horse, 2 biopsy specimens were obtained from the vastus lateralis muscle. Cryosections of equine muscle were stained with PKC isoform (alpha, beta1, beta2, delta, epsilon, or zeta)-specific polyclonal antibodies and examined by use of a fluorescence microscope. Homogenized muscle samples were evaluated via western blot analysis. RESULTS: The PKC alpha, beta1, beta2, delta, epsilon, and zeta isoforms were localized within the fibers of equine skeletal muscle. In addition, PKC alpha and beta2 were detected near or in the plasma membrane of muscle cells. For some PKC isoforms, distribution was specific for fiber type. Staining of cell membranes for PKC alpha was observed predominantly in fibers that reacted positively with myosin heavy chain (MHC)-IIa; PKC delta and epsilon staining were more pronounced in MHC-I-positive fibers. In contrast, MHC-I negative fibers contained more PKC zeta than MHC-I-positive fibers. Distribution of PKC beta1 was equal among the different fiber types. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that PKC isoforms are expressed in equine skeletal muscle in a fiber type-specific manner. Therefore, the involvement of PKC isoforms in signal transduction in equine skeletal muscle might be dependent on fiber type.     

Lingual abscesses in three dogs

Lingual abscessation is a rare condition in dogs. Very little information is available on the diagnosis and treatment of lingual abscesses in the major surgical textbooks and current veterinary literature. The common clinical signs of lingual abscesses are macroglossia, hypersalivation and a reluctance to open the mouth, but these can vary depending on the time course of the disease and the location of the abscess. This article presents three cases of tongue abscess in the dog outlining treatment and outcomes. A thorough diagnostic work up, consisting of anamnesis, clinical and haematological examinations, oral inspection under sedation and the use of diagnostic imaging techniques should be mandatory before surgical exploration of the abscess. Surgery is followed by drainage and systemic antibiotics, complemented by systemic fluid support and pain management. Conservative management of lingual abscesses can be fatal. Sharp trauma from an unknown object is suspected to be the underlying cause for the abscesses in the present cases.     

Comparison of two-dimensional and three-dimensional systems for kinematic analysis of the sagittal motion of canine hind limbs during walking

OBJECTIVE: To test the hypotheses that kinematic data of the sagittal motion of canine hind limbs during walking obtained with a 2-dimensional (2-D) system correlate well with those obtained with a 3-dimensional (3-D) system and that the data obtained with the 2-D system are repeatable. ANIMALS: 6 adult dogs with no evidence of lameness. PROCEDURES: Hind limb motions of 6 walking dogs were recorded via 2-D video and 3-D optoelectric systems simultaneously. Five valid trials were digitized, and 5 data sets (2-D 60 Hz, 3-D 180 Hz, 3-D sagittal 180 Hz, 3-D 60 Hz, and 3-D sagittal 60 Hz) of a complete gait cycle were created for each dog. In sagittal data sets, 3-D data were reduced to exclude coordinates for mediolateral orientation. Temporospatial parameters; angles of hip, stifle, and tarsal joints; and coefficients of variation of angular measurements of each dog were calculated for each data set. Accuracy of the 2-D analysis was determined by calculating mean absolute differences and estimating agreement between the 2-D and 3-D 180-Hz data sets. RESULTS: Values of joint angles and angular excursions measured with the 2-D system were repeatable and agreed with respective values obtained with the 3-D system. Reduction of the sampling rate had a greater impact on values of kinematic variables obtained with the 3-D system than did elimination of data on mediolateral orientation. CONCLUSIONS AND CLINICAL RELEVANCE: Kinematic analysis using a 2-D video system provided accurate and repeatable data of the sagittal angular motion of canine hind limbs during walking.     

Congenital malformations of the lumbosacral vertebral column are common in neurologically normal French Bulldogs,English Bulldogs,and Pugs,with breed‐specific differences

Congenital vertebral malformations are common findings on diagnostic imaging of the vertebral column in “screw‐tailed” brachycephalic dogs. The aims of this study were to evaluate the prevalence and anatomical characteristics of lumbosacral congenital vertebral malformations in French Bulldogs, English Bulldogs, and Pugs presenting for problems unrelated to spinal disease, as well as possible associations with the degree of tail malformation, lumbosacral intervertebral disc herniation, or spondylosis deformans. In this retrospective cross‐sectional study, CT scans of vertebrae L6 to S3 and of the coccygeal vertebrae were reviewed for type of congenital vertebral malformations (hemivertebrae, block vertebrae, lumbosacral transitional vertebrae, and spina bifida), lumbosacral intervertebral disc herniation, lumbosacral spondylosis deformans, and degree of tail malformation. In 76 (51.0%) of the 149 included dogs (53 French Bulldogs, 37 English Bulldogs, and 59 Pugs) at least one type of congenital vertebral malformations was found, with lumbosacral transitional vertebrae being the most common (34.2%). There was a significantly higher prevalence of lumbosacral transitional vertebrae (54.2%) and lower prevalence of hemivertebrae (1.7%) in Pugs compared to English (13.5% and 24.3%, respectively) and French Bulldogs (26.4% and 32.0%, respectively). Tail malformation was significantly more severe in dogs with evidence of hemivertebrae. Congenital vertebral malformations are a common finding in the lumbosacral vertebral column of French Bulldogs, English Bulldogs, and Pugs. These anatomical variances need to be considered when interpreting diagnostic studies and when planning for neurosurgical and neurodiagnostic procedures. Furthermore, this study suggests a possible association between the degree of tail malformation and lumbosacral hemivertebrae.     

Genetic diversity in Nordic horseradish,Armoracia rusticana,as revealed by AFLP markers

176 horseradish accessions from the Nordic countries Denmark, Finland, Norway and Sweden were analysed to estimate the level of genetic diversity and to propose conservation strategies for this cultivated plant. Most of the accessions were collected in old gardens of the Nordic countries but selections from European countries and Danish breeding lines were also included in the study. Since horseradish is mainly vegetative propagated the genetic diversity has been assumed to be small. However, using the AFLP method with three primer combinations we revealed a significant genetic diversity among Nordic horseradish. The analysis yielded 65 polymorphic bands and we found an overall diversity index of 0.5 (Shannon–Weaver). The highest diversity was found among the Finnish accessions followed by the Danish accessions. An overall AMOVA analysis indicated that 90 % of the variation could be explained by among accession variation. The AFLP data assigned the different accessions into groups that corresponded with their country of origin. A closer relationship was observed between the Swedish, Danish and some of the Norwegian accessions while the Finnish accessions separated more clearly from the other three countries. A possible explanation for the diversity is that horseradish probably has been introduced to the Nordic countries at many occasions during a long period of time.     

Vesicular stomatitis virus replicon expressing the VP2 outer capsid protein of bluetongue virus serotype 8 induces complete protection of sheep against challenge infection

Bluetongue virus (BTV) is an arthropod-borne pathogen that causes an often fatal, hemorrhagic disease in ruminants. Different BTV serotypes occur throughout many temperate and tropical regions of the world. In 2006, BTV serotype 8 (BTV-8) emerged in Central and Northern Europe for the first time. Although this outbreak was eventually controlled using inactivated virus vaccines, the epidemic caused significant economic losses not only from the disease in livestock but also from trade restrictions. To date, BTV vaccines that allow simple serological discrimination of infected and vaccinated animals (DIVA) have not been approved for use in livestock. In this study, we generated recombinant RNA replicon particles based on single-cycle vesicular stomatitis virus (VSV) vectors. Immunization of sheep with infectious VSV replicon particles expressing the outer capsid VP2 protein of BTV-8 resulted in induction of BTV-8 serotype-specific neutralizing antibodies. After challenge with a virulent BTV-8 strain, the vaccinated animals neither developed signs of disease nor showed viremia. In contrast, immunization of sheep with recombinant VP5 - the second outer capsid protein of BTV - did not confer protection. Discrimination of infected from vaccinated animals was readily achieved using an ELISA for detection of antibodies against the VP7 antigen. These data indicate that VSV replicon particles potentially represent a safe and efficacious vaccine platform with which to control future outbreaks by BTV-8 or other serotypes, especially in previously non-endemic regions where discrimination between vaccinated and infected animals is crucial.