Quantitative PCR assays to enumerate Rhizobium leguminosarum strains in soil also target non viable cells and overestimate those detected by the plant infection method

The plant infection method is commonly used to estimate the Most Probable Number (MPN) of soil rhizobia. Here, a qPCR method was set-up and validated with newly developed ANU (strain specific) and RHIZ (more general) primers to quantify the specific Rhizobium leguminosarum bv. trifolii ANU843 strain or general R. leguminosarum strains. Detection limits of qPCR protocols in soil were 1.2 × 10⁴ (ANU) and 4.2 × 10³ (RHIZ) cells per g soil. The qPCR assay appears robust and accurate in freshly inoculated soils but overestimated MPN for indigenous soil rhizobia. An incubation experiment showed that qPCR detected added DNA or non viable cells in soils up to 5 months after addition and incubation at 20 °C in moist conditions.     

Blastomycotic osteomyelitis associated with severe lameness in a horse

A 12-year-old Quarter horse gelding was presented for evaluation of severe right forelimb lameness, 2 draining tracts over the lateral aspect of the right proximal antebrachium, and weight loss. A presumptive diagnosis of blastomycotic osteomyelitis was established based on radiographs and cytology of the exudate. This diagnosis was confirmed at necropsy.     

Computed tomographic evaluation of canine radioulnar incongruence in vivo

OBJECTIVE: To compare radioulnar incongruence (RUI) of normal canine elbows and elbows with arthroscopically confirmed medial compartment disease in vivo using systematic computed tomography (CT) measurements. STUDY DESIGN: Prospective comparison of RUI measurements in normal and dysplastic canine elbows. SAMPLE POPULATION: Right elbows of 25 medium-large breed, adult dogs with medial compartment disease and 9 medium-large breed, adult dogs with no elbow disease. METHODS: Transverse CT images of proximal radioulnar articulation were reformatted to dorsal and sagittal planes. RUI in 3 locations of the forelimb's medial coronoid was measured. Arthroscopy confirmed diagnosis of medial compartment disease in the diseased group. RUI measurements of the diseased and normal elbows were compared. RESULTS: Cumulative statistical analysis of RUI in all planes revealed no significant difference between the normal and abnormal elbows (P = .61). The abnormal elbows had negative mean RUI at the mid (P = .56) and cranial (P = .24) coronoid regions that were not significantly different from normal elbows and mean positive RUI at the base coronoid that was significantly greater than in normal elbows (P = .00082). CONCLUSION: Canine elbows with established medial compartment disease do not have significant RUI at the medial coronoid region at the time of diagnosis when compared with normal elbows. CLINICAL RELEVANCE: If RUI is a significant factor in the pathophysiology of medial compartment elbow disease in the dog, it does not appear to be present at the time of diagnosis of disease. Ulnar or radial osteotomies do not appear to be indicated for restoration of normal radioulnar articular surface alignment.     

Immunological basis of differences in disease resistance in the chicken

Genetic resistance to diseases is a multigenic trait governed mainly by the immune system and its interactions with many physiologic and environmental factors. In the adaptive immunity, T cell and B cell responses, the specific recognition of antigens and interactions between antigen presenting cells, T cells and B cells are crucial. It occurs through a network of mediator proteins such as the molecules of the major histocompatibility complex (MHC), T cell receptors, immunoglobulins and secreted proteins such as the cytokines and antibodies. The diversity of these proteins that mainly is due to an intrinsic polymorphism of the genes causes phenotypic variation in disease resistance. The well-known linkage of MHC polymorphism and Marek's disease resistance difference represents a classic model revealing immunological factors in resistance differences and diversity of mediator molecules. The molecular bases in any resistance variation to infectious pathogens are vaguely understood. This paper presents a review of the major immune mediators involved in resistance and susceptibility to infectious diseases and their functional mechanisms in the chicken. The genetic interaction of disease resistance with production traits and the environment is mentioned.     

Intermittent Stress in Pigs: Behavioural and Pituitary-Adrenocortical Reactivity

Abstract

Ninetysix pigs, half females and half castrated males from 12 litters, were housed in 24 groups of 4 litter mates (2 females, 2 castrates). The pigs were weaned at 4 weeks of age and from the age of 115 days half of the groups were subjected to a schedule of unpredictable, inescapable electroshocks for 33 days, and half served as control. The adrenocortical reactivity to ACTH stimulation and to an emotional stressor (the procedure associated with initiation of blood collection) was measured in females, whereas the behavioural and pituitary-adrenocortical reactivity to an open field test was measured in castrates, as was the behavioural reactivity in a social test. Intermittent stress increased adrenocortical reactivity to ACTH stimulation significantly. This effect was present within the first week of intermittent stress (p < 0.06), but no difference was found after 4–5 weeks of stress. In contrast, the adrenocortical reactivity to additional emotional stressors was unaffected by stress after one week of intermittent stress, but increased after 4–5 weeks. In the open field test, centre location increased after one week of intermittent stress, whereas exploration decreased and walking and ambulation increased after 4–5 weeks of stress. In addition, 4–5 weeks of intermittent stress decreased aggressiveness in a social test. In conclusion, the change in reactivity to an additional acute stressor during intermittent stress appeared in different variables and contexts at different times during the course of intermittent stress. Thus, assessment of stress based on changed reactivity has to include several distinct measures of behavioural and hormonal reactivity.     

A morphological and molecular study of Anaplasma phagocytophilum transmission events at the time of Ixodes ricinus tick bite

Background

Anaplasma phagocytophilum is the causative agent of human granulocytic anaplasmosis (HGA) in humans and tick-borne fever (TBF) in ruminants. The bacterium invades and replicates in phagocytes, especially in polymorphonuclear granulocytes.

Methods

In the present study, skin biopsies and ticks (Ixodes ricinus) were collected from tick feeding lesions on 38 grazing lambs between two and three weeks after access to pastures. The histopathological changes associated with tick bites and A. phagocytophilum infection, were described. In addition the skin biopsies were examined by immunohistochemistry. Furthermore, samples from blood, skin biopsies and ticks were examined by serology, PCR amplification of msp2 (p44), genotyping of rrs (16S rRNA) variants, and compared with the results obtained from histological and immunohistochemical investigations.

Results

Tick bites were associated with chronic and hyperplastic inflammatory skin lesions in this study. A. phagocytophilum present in skin lesions were mainly associated with neutrophils and macrophages. Bacteria were occasionally observed in the Tunica media and Tunica adventitia of small vessels, but were rarely found in association with endothelial cells. PCR and genotyping of organisms present in blood, ticks and skin biopsies suggested a haematogenous and a local spread of organisms at the tick attachment sites.

Conclusions

The present study describes different aspects of A. phagocytophilum infection at the site of tick bite, and indicates that A. phagocytophilum rarely associates with endothelium during the early pathogenesis of infection.