Detection of synovial macrophages in the joint capsule of dogs with naturally occurring rupture of the cranial cruciate ligament

OBJECTIVE: To test the hypotheses that the densities of macrophages in the synovial membranes and capsules of stifle joints in dogs with ruptured cranial cruciate ligaments are greater than those of normal joints and that those densities in affected joints are positively correlated with the chronicity and severity of the disease. ANIMALS: 17 dogs with naturally occurring rupture of the cranial cruciate ligament and 5 healthy control dogs. PROCEDURE: All dogs underwent orthopedic and radiographic evaluations. In affected dogs, duration of clinical signs was used as an indicator of disease chronicity and the severity of osteoarthritis in the stifle joint was determined radiographically. Joint capsule specimens were evaluated histologically; macrophages, interleukin-6, and tumor necrosis factor-alpha were identified by use of immunocytochemical techniques. RESULTS: Compared with unaffected joints, macrophage density was increased in all affected joints. Duration of disease was significantly associated with radiographic severity of osteoarthritis and synovial macrophage density. Synovial macrophage density was significantly associated with severity of osteoarthritis and with the presence of interleukin-6 and tumor necrosis factor-a. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that synovial macrophages may be involved in the development of pathologic changes (including osteophyte formation) in the stifle joints of dogs with osteoarthritis secondary to rupture of the cranial cruciate ligament. Determination of the importance of synovial macrophages in the development of changes in osteoarthritic joints may result in new treatment strategies that involve elimination of the deleterious effects of those cells.     

Breed susceptibility for developmental orthopedic diseases in dogs

A large-scale epidemiological study was conducted to determine breeds at risk for 12 developmental orthopedic diseases (DODs). Developmental orthopedic diseases investigated included canine hip dysplasia (CHD); craniomandibular osteopathy (CMO); fragmented coronoid process; hypertrophic osteodystrophy; Legg-Calvé-Perthes disease; osteochondrosis of the medial humeral condyle, caudal humeral head, femoral condyles, and talar trochlear ridges; panosteitis; patella luxation; and ununited anconeal process. Dogs that were diagnosed with any one of the diseases of interest at any of 10 veterinary teaching hospitals participating in the Veterinary Medical Database from 1986 to 1995 were included as cases. Odds ratios and corresponding 95% confidence intervals were calculated to determine risk. Frequency of diagnosis during the 10-year period ranged from 35 cases (CMO) to 10,637 cases (CHD). The number of breeds at increased risk for a disease ranged from one (CMO) to 35 (CHD). Breed susceptibility for a DOD may suggest a genetic component in the disease etiology. The results of this study serve to increase veterinarians' awareness of breeds susceptible to DODs and may facilitate the control of such diseases by identifying breeds that might benefit from breeding programs or environmental intervention such as dietary modification.     

A Gene-for-Gene Relationship Between Wheat and Mycosphaerella graminicola, the Septoria Tritici Blotch Pathogen

ABSTRACT Specific resistances to isolates of the ascomycete fungus Mycosphaerella graminicola, which causes Septoria tritici blotch of wheat, have been detected in many cultivars. Cvs. Flame and Hereward, which have specific resistance to the isolate IPO323, were crossed with the susceptible cv. Longbow. The results of tests on F1 and F2 progeny indicated that a single semidominant gene controls resistance to IPO323 in each of the resistant cultivars. This was confirmed in F3 families of Flame x Longbow, which were either homozygous resistant, homozygous susceptible, or segregating in tests with IPO323 but were uniformly susceptible to another isolate, IPO94269. None of 100 F2 progeny of Flame x Hereward were susceptible to IPO323, indicating that the resistance genes in these two cultivars are the same, closely linked, or allelic. The resistance gene in cv. Flame was mapped to the short arm of chromosome 3A using microsatellite markers and was named Stb6. Fifty-nine progeny of a cross between IPO323 and IPO94269 were used in complementary genetic analysis of the pathogen to test a gene-for-gene relationship between Stb6 and the avirulence gene in IPO323. Avirulence to cvs. Flame, Hereward, Shafir, Bezostaya 1, and Vivant and the breeding line NSL92-5719 cosegregated, and the ratio of virulent to avirulent was close to 1:1, suggesting that these wheat lines may all recognize the same avirulence gene and may all have Stb6. Together, these data provide the first demonstration that isolate-specific resistance of wheat to Septoria tritici blotch follows a gene-for-gene relationship.     

Multi-scale feedbacks between tree regeneration traits and herbivore behavior explain the structure of pasture-woodland mosaics

Context

The pasture-woodlands of Central Europe are low-intensity grazing systems in which the structural richness of dynamic forest-grassland mosaics is causal for their high biodiversity. Distinct mosaic patterns in Picea abies- and Fagus sylvatica-dominated pasture-woodlands in the Swiss Jura Mountains suggest a strong influence of tree species regeneration ecology on landscape structural properties. At the landscape scale, however, cause-effect relationships are complicated by habitat selectivity of livestock.

Objectives

We asked which tree species regeneration traits and what kind of feedbacks among local-scale vegetation dynamics and landscape-scale herbivore behavior are causal for the contrasted landscape structural characteristics of Picea- and Fagus-dominated pasture-woodlands.

Methods

We performed simulation experiments of mosaic pattern formation in both pasture-woodland types. The regeneration traits, namely dispersal distance, resistance to browsing and tolerance to shade, and the rules for habitat selection of cattle were modified and the corresponding shifts in landscape structure were analyzed.

Results

Dispersal distance showed a significant, but only local, effect promoting forest fringe formation. Saplings’ resistance to browsing mainly determined overall tree cover, but did not influence landscape structure. At the landscape scale, both shade tolerance of saplings and selective habitat use by cattle were responsible for forest-grassland segregation: high shade tolerance triggered segregation, whereas non-selective habitat use hindered it.

Conclusions

Existing local-scale theory on pasture-woodland dynamics is complemented by an herbivore-vegetation feedback among spatial scales. In low-intensity pastures, where large herbivores are preferentially “grazers” and trees form dense canopies, an intrinsic trend towards forest-grassland segregation at the expense of forest-grassland ecotones is predicted.

     

Comparative measurements of resistance to insecticides in three closely-related old and new world bollworm species

Toxicity measurements were made with three closely-related Bollworm species, Heliothis virescens F., Helicoverpa zea (Boddie), and Helicoverpa armigera (Huebner) using insecticides of 3 different chemical classes. Tests were carried out by a leaf-dip method using neonate larval stages, a topical application test on 3rd larvae, and a topical test with imagines. H. armigera populations from Ivory Coast, Turkey, Thailand, and India were obtained and investigated. H. virescens originated from the USA and Colombia, and H. zea from Guatemala and Nicaragua. Resistance was expressed in all species, the sensitive reference strains having similar LC₅₀s in tests with monocrotophos, profenofos, thiodicarb, and methomyl. Since resistance appeared not to be a quality related to species but rather a feature common to all of the samples investigated, its origin must be connected with the type of cotton culture and the selection pressure they were subjected to in their respective agro-ecosystem. Resistance was highest for monocrotophos and cypermethrin in Central American countries, differences in the treatment regime being reflected by the resistance levels attained. In all stages tested, resistance was sufficiently well expressed to make a monitoring system based on neonate larvae, the 3rd larval stage, or imagines of the species possible. The final decision as to which should be used for a general, comparative monitoring system, therefore, should be governed by the technical parameters of the testing process.     

Quantitative Detection of Fusarium Species in Wheat Using TaqMan

Fusarium head blight (FHB) of wheat and other small-grain cereals is a disease complex caused by several fungal species. To monitor and quantify the major species in the FHB complex during the growing season, real-time PCR was developed. TaqMan primers and probes were designed that showed high specificity for Fusarium avenaceum, F. culmorum, F. graminearum, F. poae and Microdochium nivale var. majus. Inclusion of an internal PCR control and serial dilutions of pure genomic DNAs allowed accurate determination of the concentration of fungal DNA for each of these species in leaves, ears as well as harvested grains of winter wheat. The DNA concentration of F. graminearum in grain samples correlated (r ²= 0.7917) with the incidence of this species on the grain as determined by isolation from individual kernels. Application of the TaqMan technology to field samples collected in 40 wheat crops in the Netherlands during the growing season of 2001 revealed that M. nivale var. majus predominated on leaves early in the season (GS 45-65). Ears and harvested grains from the same fields, however, showed F. graminearum as the major species. In 2002, grain samples from 40 Dutch fields showed a much wider range of species, whereas in ears from 29 wheat crops in France, F. graminearum was the predominant species. The concentration of DON correlated equally well with the incidence of the DON-producing species F. culmorum and F. graminearum in the grain samples (r ²= 0.8232) as well as with total DNA of both these species (r ²= 0.8259). The Fusarium TaqMan technology is an important tool to quantify and monitor the dynamics of individual species of the complex causing FHB in cereals during the growing season. This versatile tool has been applied in a comparison of different genotypes, but can also be applied to other disease management systems, e.g. fungicide treatments.     

Synteny in Toxigenic Fusarium Species: The Fumonisin Gene Cluster and the Mating Type Region as Examples

A comparative genomic approach was used to study the mating type locus and the gene cluster involved in toxin production (fumonisin) in Fusarium proliferatum, a pathogen with a wide host range and a complex toxin profile. A BAC library, generated from F. proliferatum isolate ITEM 2287, was used to identify chromosomal regions flanking the mating type locus and the gene cluster involved in the biosynthesis of fumonisin. These regions were sequenced and compared with corresponding sequences in other ascomycetes. The results demonstrated that the level of synteny between ascomycetes can vary greatly for different genomic regions and that the level of similarity of genes within a region can also fluctuate strongly. Synteny was found in the regions flanking the mating type idiomorph among ascomycetes that supposedly diverged 280 million years ago. The fumonisin gene clusters of F. proliferatum and F. verticillioides were completely syntenic but absent in F. graminearum. The regions flanking the fumonisin gene clusters were highly dissimilar between F. proliferatum and F. verticillioides, whereas they formed a continuous region in F. graminearum. This indicates that the fumonisin gene cluster has been inserted at different genome locations in both species. Surprisingly low similarity was found between the corresponding genes within the fumonisin cluster of F. proliferatum and F. verticillioides, compared to other genomic sequences indicative for two independent acquisition events from distinct genetic sources. The results demonstrate the power of comparative genomics for gene annotation and for studies on the evolution of genes, gene-clusters and species.     

Ochroconis globalis infecting Atlantic salmon (Salmo salar), with a review of Ochroconis species in cold‐blooded animals

Necropsy examination of an adult Atlantic salmon (Salmo salar) from the Dalälven River in Sweden revealed numerous large, white nodules, with spherical cysts and granulomata in kidney and liver. Histopathology showed dark, septate, thin‐walled hyphae. The aetiologic agent was found to be an Ochroconis species (Venturiales) that differed from known fish‐associated species of the genus. Molecular phylogenetic studies of the culture (strain UIII09 = CBS 135766) demonstrated that Ochroconis globalis was concerned. The isolate proved to be susceptible to all investigated antifungals, as it is known for another Ochroconis species. The role of Ochroconis in opportunism of cold‐blooded animals was discussed, and the diagnostic methods using DNA sequences for routine identification of the fungus were proposed.