Exogenous induction of cerebral beta-amyloidogenesis is governed by agent and host

Protein aggregation is an established pathogenic mechanism in Alzheimer's disease, but little is known about the initiation of this process in vivo. Intracerebral injection of dilute, amyloid-beta (Abeta)-containing brain extracts from humans with Alzheimer's disease or beta-amyloid precursor protein (APP) transgenic mice induced cerebral beta-amyloidosis and associated pathology in APP transgenic mice in a time- and concentration-dependent manner. The seeding activity of brain extracts was reduced or abolished by Abeta immunodepletion, protein denaturation, or by Abeta immunization of the host. The phenotype of the exogenously induced amyloidosis depended on both the host and the source of the agent, suggesting the existence of polymorphic Abeta strains with varying biological activities reminiscent of prion strains.     

Experimental infection of cattle, sheep and pigs with 'Hobi'-like pestivirus

To date, limited information is available on the ability of 'Hobi'-like pestiviruses (putative bovine viral diarrhoea 3) to infect and cause disease in animal species traditionally affected by pestiviruses. In order to obtain new insights into host range and pathogenic potential of this atypical pestivirus, BVDV-seronegative calves (n=5), lambs (n=5) and piglets (n=5) were experimentally infected with the European 'Hobi'-like strain Italy-1/10-1, whereas two animals per species served as uninfected controls. Appearance of clinical signs, leukopenia, viremia, viral shedding and seroconversion were monitored for 28 days post-infection. Calves and lambs were successfully infected, displaying respiratory signs (nasal discharge), moderate hyperthermia and leukopenia, viremia and viral shedding through the nasal and faecal routes. Antibody responses were observed in both animal species by ELISA and virus neutralisation assays. In contrast, inoculated piglets did not display any clinical signs nor leukopenia and viral RNA was not detected in any biological samples. Nevertheless, the presence of detectable antibodies by virus neutralisation accounted for a successful, albeit limited infection of these animals.     

Perception of dogs’ stress by their owners

Questionnaires potentially have a broad applicability in measuring stress levels in dogs, as owners know their dogs’ behavior and personality better than anyone else. The aim of this research was to evaluate how owners perceive stress in their dogs through understanding of displayed behaviors.The survey was carried out using 1,190 questionnaires completed by dog owners. More than half of respondents were able to correctly identify stress as a short- or long-term alteration of the psychophysical equilibrium that can develop into illness. This ability was higher with higher educational levels. The behavioral indicators of stress most frequently identified by owners were trembling and whining, followed by aggressiveness, excessive barking, and panting. More subtle behaviors such as looking elsewhere, turning head, yawning, and nose licking were more rarely reported, suggesting that few owners are able to correctly interpret and intervene in early stages of stress. The vast majority of respondents indicated that dogs were stressed seldom or only in specific situations. Men generally considered their dogs as experiencing low stress more often than did women, whereas women considered their dogs as being moderately stressed more often than did men.An owner’s ability to recognize behavioral signs of stress is important, as it enables the owner to help the animal to avoid welfare problems, such as stressful situations, and favors a rapid recovery of psychophysical homeostasis by interrupting the progression to overstress and distress. The results show that some owners can help in protecting the welfare of their dogs, but that many owners would benefit from educational efforts to improve their ability to interpret their dogs’ behavior.     

GeneScanning analysis of Ig/TCR gene rearrangements to detect clonality in canine lymphomas

The diagnosis of canine lymphoma is achieved using morphological and immunological methods. In a certain percentage of cases, difficulties in making a definitive diagnosis of lymphoproliferative disorders may occur despite extensive immunophenotyping. Therefore, additional diagnostics, such as molecular assessment of Ig/TCR gene rearrangements clonality, may confirm the final diagnosis. Polyacrylamide gel electrophoresis and heteroduplex analysis have already been proven to be suitable for detecting clonality but are cumbersome and labor-intensive. In the present study, GeneScanning analysis of PCR products originating from different primer sets targeting different regions of Ig and TCR was validated in improving sensitivity as well as in reducing the turnaround time of gene rearrangement assays. GeneScanning exploits 5' fluorescently labelled primers for the automated and fast analysis of PCR products either as singleplex or multiplex runs. Initially, the assay was set up using DNA purified from normal tissues (n=6), hyperplastic/reactive tissues (n=10) and a small set of immunophenotyped lymphoma samples (n=12). The optimized methods were then used in a large set of 96 canine lymphoma samples. Normal and hyperplastic/reactive lymphoid tissues showed typically polyclonal or, occasionally, oligoclonal PCR products. Lymphoma samples showed monoclonal peaks arranged as a single or, occasionally, a double narrow base peak sometimes embedded in a polyclonal background. In all immunophenotyped cases, an Ig or TCR clonal finding corresponded to B- and T-cell lymphomas, respectively. Overall, 94/96 (97.9%) samples showed clonal Ig/TCR clonal rearrangements among which clonal Ig was found in 61/96 (63.5%) of samples and clonal TCR in 33/35 Ig negative samples (34.4% of all cases). In one out of ten randomly chosen cases, both Ig and TCR clonal gene rearrangements were found. Among the factors affecting assay accuracy, DNA quality has been shown to be critical and the amplification of DNA controls of different size are recommended to evaluate DNA integrity. Frozen material such as that which remained inside the hub of the needle used for diagnostic procedures is optimal for the analysis herein described. In conclusion, GeneScanning represents a versatile tool for routinely assessing Ig/TCR clonal rearrangements and supporting the diagnostic protocol of canine lymphomas.     

Novel swine influenza virus subtype H3N1 in Italy

To date, three subtypes of swine influenza viruses, H1N1, H1N2, and H3N2 have been isolated in Italy. In 2006, a novel swine influenza virus subtype (H3N1) was isolated from coughing pigs. RT-PCR performed on lung tissues, experimental infection in pigs with the novel isolate, and cloning the virus by plaque assay confirmed this unique H and N combination. The novel isolate was also antigenically and genetically characterized. Genetic and phylogenetic analysis showed that the complete HA gene of the H3N1 strain has the highest nucleotide identity to three Italian H3N2 strains, one isolated in 2001 and two in 2004, whereas the full length NA sequence is closely related to three H1N1 subtype viruses isolated in Italy in 2004. The remaining genes are also closely related to respective genes found in H1N1 and H3N2 SIVs currently circulating in Italy. This suggests that the novel SIV could be a reassortant between the H3N2 and H1N1 SIVs circulating in Italy.     

How Do Horses Appraise Humans' Actions? A Brief Note over a Practical Way to Assess Stimulus Perception

Heart rate (HR) is considered to be an effective tool for assessing animals' emotional response to a stimulus. We investigated changes in HR during a series of handling procedures (grooming test) in horses that had different experiences of human interaction. We used four groups of horses: grazing horses (group A), school horses (group B), six ponies, traditionally trained (group C), and six ponies, trained with modulated stimulus intensity (group D). An HR monitor was applied to each horse. The operator began the grooming test, standardized at the following points: (1) trainer's entry into the enclosure/paddock, (2) halter fastened, (3) start of grooming, (4) end of grooming, (5) saddling, and (6) inserting the bit. Group A repeated the grooming test twice: group A1 with a known operator and group A2 with an unknown operator. The data were compared by using analysis of variance (ANOVA) tests. The results showed a significant increase in HR (P ≤ .05) at point 2 compared with baseline in all groups except group D. Other significant differences were found at all points between groups A1 and A2, groups A2 and B, groups A1 and C, groups B and C, and at points 5 and 6 between groups A2 and C, as well as between groups C and D. Even when a scientific assessment of the increase in the HR is of primary importance, the use of this physiologic parameter may be helpful in assessing the horse's perceptions of stimuli presented by people during handling. Our results show that it is not the action “per se” that is important, but the manner in which horses perceive and appraise such actions in relation to the environment and their subject's experiences.     

Calcium fertilizers induce soil suppressiveness to <Emphasis Type="BoldItalic">Phytophthora cinnamomi</Emphasis> root rot of <Emphasis Type="BoldItalic">Quercus ilex</Emphasis>

Based on the observation that the root disease caused by P. cinnamomi on Q. ilex has a low incidence and severity in soils with medium-high Ca²⁺ content, we studied the ability of Ca²⁺ fertilizers to induce soil suppressiveness to the pathogen. Studies on cultures of P. cinnamomi exposed to different Ca²⁺ fertilizers in vitro showed significant inhibition of sporangial, chlamydospore and zoospore production at millimolar concentrations while mycelial growth was mainly unaffected. Experiments performed with artificially infested soil showed that some Ca²⁺ fertilizers induce a significant decrease on chlamydospore viability. Additionally, greenhouse experiments using artificially infested soils showed a significant reduction of foliar and root symptom severities in Holm oak seedlings growing in soils amended with Ca²⁺ fertilizers. We suggest that limestone amendments in oak rangelands could enhance the suppressiveness of soils to P. cinnamomi, and it is likely that the inhibition of sporangial production was the main mechanism involved.