The Brain of the Dog in Section: a Comprehensive View for Veterinary Students

Transversal, horizontal and sagittal sections of the brain were stained by the ancient but efficient Mulligan method, a procedure that establishes a clear macroscopic difference between the white and grey substances. Different structures of each section were studied and most of the details were identified and named according to the NAV. All sections were projected onto the whole brain. By means of this easy and basic procedure the students increase their understanding of (1) the size and/or the form and/or the topography of several prominent structures of the brain, (2) the general distribution of the substancia alba and grisea, and they begin to understand the complexity of the brain.     

THE EFFECTS OF PATIENT POSITIONING AND SLICE SELECTION ON CANINE ACETABULAR ANGLE ASSESSMENT WITH COMPUTED TOMOGRAPHY

Acetabular angles (AAs) and dorsal acetabular rim angles acquired by computed tomographic (CT) imaging have been used to assess patient response to juvenile pubic symphysiodesis surgery. The purpose of this study was to evaluate the effects of patient positioning and slice selection on these angles, and an attempt was made to devise a repeatable method of measuring these angles that would eliminate positioning effects. We found significant variation in AAs with small differences in pelvic tilt and slice selection. Dorsal acetabular rim angles were not affected. As a result of positioning differences from one CT study to the next, every attempt should be made to standardize pelvic tilt, or eliminate its effect on AAs by standardizing gantry angle in relation to an anatomic landmark that will not change over time. The floor of the sacral vertebral canal may be a reasonable landmark for this purpose and deserves further study.     

MAGNETIC RESONANCE IMAGING FINDINGS IN SPINAL CORD INFARCTION IN THREE SMALL BREED DOGS

Fibrocartilaginous embolization (FCE) of the spinal cord is a common disease in large breed dogs. There are only a few reports about this entity in small breed dogs and it has never been reported in chondrodystrophic breed. For definitive diagnosis histopathologic examination is necessary. Magnetic resonance imaging (MRI) as a potential diagnostic tool for intravitam diagnosis of FCE has been mentioned before, but results have not been reported so far. This report describes the neurological findings and MRI results in three small breed dogs, including a Pekingese dog, with FCE of the spinal cord. The disease was suspected in two animals based upon clinical and MRI-appearance and confirmed in the third by histopathological examination. In all three cases, similar focal intramedullary lesions, consisting of hyperintensive signals on T2-weighted images, were detected. Based on these findings, high-field MRI may be used as an antemortem tool for the diagnosis of FCE. It is also shown that FCE can occur in chondrodystrophic dogs.     

Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Secondary or Accessory Glomerular Layer in Mice During Growth

Most mammals have two different structures in which we found glomerular layers at the same time: the main olfactory bulb (MOB) and the accessory olfactory bulb (AOB). Both bulbs have the same pattern of organization, but there are some differences: although the size is considerably bigger in MOB than in AOB, probably the most important difference is that the principal cells are not differentiated into mitral and tufted cells in the AOB, and are usually described as mitral/tufted cells. We have previously observed that in some mammals, like pigs and sheep, the AOB reaches maturity before birth, but this is not a rule for other species. Surprisingly, mice need several days of life to achieve full stratification of its cellular components. We have studied the chronology of this process, focusing our attention on the glomerular layer, the last to appear. We concluded that there are two critical periods, between E11.5 and E16.5 (migration phase) and between E17.5 and P3.5–7 (true morphological constitution).     

Genetic diversity in populations of the fungi Phaeomoniella chlamydospora and Phaeoacremonium aleophilum on grapevine in France

Isolates of Phaeomoniella chlamydospora ( Phc ) and Phaeoacremonium aleophilum ( Pha ), two haploid, deuteromycetous fungi, were obtained from vines showing symptoms of esca disease in different localities in two French regions, and within a single vineyard in one of these regions. The population genetic structure was determined in both fungi using random amplified polymorphic DNA (RAPD) analysis. Populations of Phc showed similar levels of diversity at local and regional levels. The most frequent Phc haplotypes were found in every population, and the frequencies of positive alleles of markers were similar across populations. The hypothesis that recombination had occurred was rejected for the full set of samples, but not for the samples reduced to haplotypes, indicating that Phc may be a recombining species. Different features were identified in Pha populations. First, the southern population of Pha appeared more diverse than the south-western populations. Second, genetic differentiation was identified between Pha populations from southern and south-western regions for several RAPDs. Finally, in the southern population of Pha no evidence for recombination was obtained, even by reducing the sample to haplotypes. Within the single vineyard surveyed, several haplotypes of both fungi were recovered and randomly distributed. Thus different infection events appeared to have occurred on a low spatial scale. Data from this study showed that haplotypes of both fungi were distributed over long distances geographically, and that most of the vineyards surveyed were infested by more than one haplotype of Phc and Pha .     

Biological and sequence comparisons of Potato virus Y isolates associated with potato tuber necrotic ringspot disease

The biological and molecular relationships between a large number of Potato virus Y (PVY) isolates were examined, concentrating mainly on isolates associated with potato tuber necrotic ringspot disease (PTNRD). Following detailed analysis of the coat-protein gene, four main groups were identified which broadly corresponded to the phenotype of the different isolates. The groups comprised the ordinary strain (PVY^O), the necrotic strain (PVY^N), the C strain (PVY^C) and a group of recombinant (between ordinary and necrotic) isolates. In the latter group, all members were associated with PTNRD. However, four nonrecombinant isolates were also identified which were associated with PTNRD or tuber necrosis. Three were from tubers showing PTNRD symptoms in the field, while the fourth originated from symptomless tubers, but could cause necrotic rings on tubers under glasshouse conditions. The results show that although coat-protein recombination is always found associated with the PTNRD phenotype, some nonrecombinant isolates have very similar biological properties.     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

Evaluation of resistance to Phytophthora cinnamomi in seed-grown trees and clonal lines of Eucalyptus marginata inoculated in lateral branches and roots

Seed-grown trees and six clonal lines of 3·5–4·5-year-old Eucalyptus marginata (jarrah) growing in a rehabilitated bauxite mine site in the jarrah forest were underbark-inoculated on lateral branches (1995) or simultaneously on lateral branches and lateral roots (1996) with isolates of Phytophthora cinnamomi in late autumn. Individual seedlings from which the clonal lines were derived had previously been assessed as either resistant (RR) or susceptible (SS) to P. cinnamomi . At harvest, the acropetal lesion and colonization lengths were measured. Overall, the length of colonization in roots and branches was more consistent as a measure of resistance than lesion length, because colonization length recorded the recovery of P. cinnamomi from macroscopically symptomless tissue ahead of the lesion which, on some occasions, was up to 6 cm. In both trials, one RR clonal line was able to contain the P. cinnamomi isolates consistently, as determined by small lesion and colonization lengths in branches and roots. In contrast, the remaining two RR clonal lines used in both trials were no different from the SS line in their ability to contain lesions or colonization. These latter two RR lines may therefore not be suitable for use in rehabilitation of P. cinnamomi -infested areas. Differences in lesion and colonization lengths among P. cinnamomi isolates occurred only in the 1995 trial. Colonization and lesion lengths in branches were up to eight times greater in 1996 than in 1995, but the relative rankings of clonal lines were consistent between trials. Although colonization was always greater in branches than roots, the relative rankings of the lines were similar between branch and root inoculations. Branch inoculations are a valid option for testing the resistance and susceptibility of young jarrah trees to P. cinnamomi .     

Identification and characterization of Pepino mosaic potexvirus in tomato

At the beginning of 1999, a new virus disease occurred in protected tomato crops in The Netherlands. Initial diagnostic tests revealed the presence of a potexvirus but serological tests ruled out the presence of Potato X potexvirus (PVX). Tests for other potexviruses reported from solanaceous crops provisionally identified the virus as Pepino mosaic potexvirus (PepMV). The virus was purified, and an antiserum was produced, which showed strong reactions with both the type isolate of PepMV from pepino and two other isolates from tomato. Host range and symptomatology of the pepino and tomato isolates of PepMV revealed clear differences from PVX. However, differences were also observed between the pepino and tomato isolates of PepMV. Sequence alignment of DNA fragments of 584 bp derived from the RNA polymerase cistron showed almost 95% identity with the pepino isolate, whereas the identity with PVX appeared to be < 60%. Together, these results identified PepMV as the causal agent of the new virus disease in tomato. Based on the differences from the type isolate from pepino ( Solanum muricatum ), the isolates from tomato should be considered as a distinct strain of PepMV for which the name tomato strain is proposed.