Cloning and expression of pigeon IFN-γ gene

This is the first paper describing the cloning of pigeon IFN-γ gene (PiIFN-γ) and the analysis of the in vitro expressed recombinant protein. The PiIFN-γ gene was identified by RT-PCR as a 498 bp, fragment coding for a precursor protein of 165 amino acids instead of 164 amino acids, as observed in the other avian species. The recombinant protein was expressed in vitro by an eukaryotic system and the biological properties of the cytokine were tested using a chicken macrophage cell line. The high degree of amino acid and nucleotide identity, shared with the ChIFN-γ, and the fact that the pigeon protein was functional on chicken cells, indicates a cross-reactivity between pigeon and chicken IFN-γ. The detection of the PiIFN-γ could represent an useful instrument in understanding the role played by this cytokine in immune response related to vaccinations and infectious diseases in the pigeon.     

Squash-prep cytology in the diagnosis of canine and feline nervous system lesions: a study of 42 cases

BACKGROUND: The increased sophistication of imaging techniques in veterinary medicine allows the detection of a wide variety of intracranial and intraspinal lesions; however, imaging often does not provide a definitive diagnosis for nervous system (NS) lesions. Cytology is emerging as a useful diagnostic tool for obtaining a fast and accurate assessment of NS lesions, but little information is available for dogs and cats. OBJECTIVES: The purpose of this study was to assess the accuracy of cytologic evaluation of squash samples from NS lesions in dogs and cats and to consider cytology-based diagnostic guidelines and sources of misdiagnosis. METHODS: Cytologic specimens from masses localized in the central and peripheral NS taken during surgery or postmortem examination were classified into 3 groups according to the final histopathologic diagnosis: Group 1 = completely correct diagnosis, when the cytologic diagnosis and final histologic diagnosis were exactly correlated; Group 2 = partial correlation, when the cytologic diagnosis only partially correlated with the final histologic diagnosis, and Group 3 = no correlation, when the cytologic diagnosis was incorrect and there was no correlation with the general histologic type of lesion. The diagnostic accuracy of cytopathology was calculated by considering the histopathologic diagnosis as the "gold standard," and calculating a 95% confidence interval (CI). RESULTS: A total of 42 animals (33 dogs and 9 cats) were included in the study. The cytologic diagnoses were classified in Group 1 for 32 cases (76%; 95% CI 0.63-0.89), in Group 2 for 6 cases (14%; 95% CI 0.04-0.25), and in Group 3 for 4 cases (10%; 95% CI 0.006-0.18). Considering both complete and partial correlation as an adequate result, cytologic diagnosis was satisfactory in 90% of biopsies. CONCLUSIONS: Although the current series of cases is relatively small, cytologic evaluation of squash preparations can be considered a fairly accurate and reliable tool in the diagnosis of NS lesions.     

Multinucleated giant cells with an osteoclast phenotype derived from caprine peripheral blood mononuclear cells

Formation of multinucleated giant cells (MGCs) by macrophage fusion is a typical cytopathic effect of lentiviral replication in caprine monocytes and MGC formation from cultured caprine peripheral blood mononuclear cells (PBMCs) has been considered to be diagnostic for small ruminant lentivirus (SRLV) infection. In this study, formation of MGCs was observed after 7–14 days when PBMCs were cultured from healthy goats free from SRLV infection. These MGCs expressed tartrate-resistant acid phosphatase, calcitonin receptor, integrin αVβ3, cathepsin K and matrix metalloproteinase 9 and were able to resorb bone in vitro in the absence of RANKL and macrophage colony stimulating factor, consistent with an osteoclast phenotype.     

Influence of season on the hematological and serum biochemical values of bottlenose dolphins (Tursiops truncatus) housed in a controlled environment in northern Italy

Season appears to influence the physiology of bottlenose dolphins (Tursiops truncatus). From 2000 to 2005, blood parameters of three captive bottlenose dolphins housed in northern Italy were examined to determine if seasonal variation was present. Seasonal variation was observed in the male dolphins, with both males exhibiting aminotransferase levels that were higher in autumn and lower in winter. Mean serum creatinine levels were higher during summer and lower during autumn in the adult male, and mean lactate dehydrogenase higher during summer and lower during spring in the juvenile male. Both males exhibited red blood cell counts and hemoglobin levels that were higher during autumn and lower during summer. This study contributes to the knowledge of baseline hematologic and biochemical values based on seasonality in bottlenose dolphins.     

Genetic and antigenic characterization of the surface lipoprotein P48 of Mycoplasma bovis

The presence of a membrane lipoprotein homologous to the P48 of Mycoplasma agalactiae was investigated in different Mycoplasma bovis isolates selected by geographical locations and biological properties. Its potential as a diagnostic tool was also discussed. The presence of a specific signal observed in all M. bovis field isolates probed with a rabbit antiserum raised against the M. agalactiae recombinant P48 demonstrated that this protein is structurally and antigenically conserved within the M. bovis cluster. No signal was detected when testing six different mycoplasma species found in cattle. The p48 gene was identified by PCR approach and partially sequenced. Full length gene sequence was obtained by direct bacterial chromosome sequencing. Five UGAs were selectively mutated into UGG and the full length mutated gene, lacking the signal peptide, was cloned and expressed in Escherichia coli. The purified recombinant antigen (r-P48) was evaluated as a potential marker of infection using a panel of 86 well-characterized sera from experimentally and naturally infected cattle. Specific IgM antibodies were detected within 6-9 days after experimental infection followed by an IgG response lasting from the third/fourth week after contact. Although antibody titers were well below those observed in sheep or goats infected with M. agalactiae, results suggest that M. bovis r-P48 can be used as a specific marker of infection.     

FOAM, a new simple benthic degradative module for the LAMP3D model: an application to a Mediterranean fish farm

The modelling framework already introduced by Doglioli, Magaldi, Vezzulli and Tucci to predict the potential impact of a marine fish farm is improved following different directions, namely (1) real historic current-metre data are used to force the simulations, (2) settling velocity values specifically targeting Mediterranean fish species are used, and (3) a new benthic degradative module, the Finite Organic Accumulation Module, is added to the modelling framework. The Finite Organic Accumulation Module uses the output of the other functional units of the modelling framework to calculate the organic load on the seabed. The Finite Organic Accumulation Module considers the natural capability of the seafloor in absorbing part of the organic load. Different remineralization rates reflect the sediment stress level according to the work of Findlay and Watling. Organic degradation for both uneaten feed and faeces is evaluated by changing the release modality (continuous and periodical) and by varying the settling velocities. It is found that the maximum impact on the benthic community is observed either for quickly sinking uneaten feed released twice a day, or for less intense near-bottom current conditions. If both the above-mentioned scenarios coexist, a high stress level is established in the sediment. The model also suggests that the use of self-feeders in cages can reduce farm impacts significantly. These results show how the new and more complete modelling framework presented here is able to improve the objectivity in the decision-making processes and how it may be successfully used for planning and monitoring purposes.     

Morphology of the Atlantic salmon (Salmo salar) tongue

The Atlantic salmon (Salmo salar) is a freshwater and marine fish of the family Salmonidae, widely farmed in aquaculture facilities in several countries. The salmon are carnivorous, but in aquaculture, alternative foods have been experienced. It is well known that feeding in captivity should cause adaptation and modifications of the morphological characteristics of the oral cavity, especially of tongue; therefore, the aim of this study was to investigate, by light, laser confocal and scanning electron microscopy, the morphological characteristics of the tongue dorsal surface, considering the importance of the correlations between feeding habits and the anatomy of the tongue. Scanning electron microscopy demonstrates the presence of caniniform teeth with oro-aboral orientation surrounded by numerous filiform papillae, single, fused or arranged in row. Oro-aborally, the papillae show an appearance like a rosette and they disappear at level of the root. Light and laser confocal microscopy demonstrates that the mucosa is covered by a non-keratinized stratified pavement epithelium with, in the deepest layer, the presence of a triangular structure whose apex is cranially directed and base facing aborally. In this structure, spindle-shaped cells are present, with a vimentin immunoreactivity, that for their characteristics could be adult mesenchymal stem cells. The obtained data could be useful not only for further studies on the nutrition, but it is interesting the detection of tissues typical of the embryo-fetal phase in the adult specimens tongue, thus giving a basis for studies of potential applications, if any, regarding cell therapies for different clinical indications.     

Detection of Helicobacter spp. in gastric, fecal and saliva samples from swine affected by gastric ulceration

The aim of this study was to evaluate the presence of Helicobacter (H.) spp. in swine affected by gastric ulceration. Stomachs from 400 regularly slaughtered swine were subjected to gross pathological examination to evaluate the presence of gastric ulcers. Sixty-five samples collected from ulcerated pars esophagea and 15 samples from non-ulcerated pyloric portions were submitted to histopathological and molecular analyses, to detect Helicobacter spp., H. suis and H. pylori by PCR. Feces and saliva swabs were also collected from 25 animals in order to detect in vivo the presence of Helicobacter spp.. Gastric ulcers were detected in 373 cases (93%). The presence of ulcers in association with inflammatory processes was further confirmed by histological examination. Forty-nine percent (32/65) of the ulcerated esophageal portions as well as 53% (8/15) of the non-ulcerated pyloric portions were positive for Helicobacter spp. by PCR. The Helicobacter spp. positive samples were also positive for H. suis, while H. pylori was not detected. These results were confirmed by restriction enzyme analysis. With regard to feces and saliva samples, 15/25 (60%) and 16/25 (64%) were positive for Helicobacter spp. PCR, respectively but all were negative in H. suis and H. pylori specific PCR.     

Phenotypic and genotypic characterization of canine pyoderma isolates of Staphylococcus pseudintermedius for biofilm formation

Biofilm-forming ability is increasingly being recognized as an important virulence factor in several Staphylococcus species. This study evaluated the biofilm-forming ability of sixty canine derived clinical isolates of S. pseudintermedius, using three phenotypic methods, microtiter plate test (MtP), Congo red agar method (CRA) and tube adherence test, and the presence and impact of biofilm-associated genes (icaA and icaD). The results showed that icaA and icaD genes were detected concomitantly in 55 (91.7%) of 60 isolates. A majority (88.3%) of the strains screened had matching results by the tube adherence test, MtP and PCR analysis. Better agreement (95%) was found between the PCR-based analysis and the CRA. Results of the icaA and icaD gene PCRs showed good agreement with CRA results, with a kappa of 0.7. Comparing the phenotypic methods, the statistical analysis showed that the agreement among the phenotypical tests using categorical data was generally good. Considering two classes (biofilm producer and biofilm non-producer), the percentage of matching results between the CRA method and the tube adherence test and between the CRA method and the MtP was 93.3%. A concordance of 100% was revealed between the MtP and the tube adherence test. The results indicate a high prevalence of the ica genes within S. pseudintermedius isolates, and their presence is associated with in vitro formation of a biofilm. A combination of phenotypic and genotypic tests is recommended for investigating biofilm formation in S. pseudintermedius.     

Hepatic encephalomyelopathy in a calf with congenital portosystemic shunt (CPSS)

A 4-month-old female Holstein Friesian calf was referred to the Veterinary Teaching Hospital, University of Berne, Switzerland for evaluation of ataxia, weakness, apathy and stunted growth. Clinical examination revealed generalized ataxia, propioceptive deficits, decreased menace response and sensibility. Postmortem examination did not reveal macroscopic changes of major organs. Histologically, the brain and the spinal cord lesions were characterized by polymicrocavitation, preferentially affecting the white matter fibers at the junction of grey and white matter and by the presence of Alzheimer type II cells. The liver revealed lesions consistent with a congenital portosystemic shunt, characterized by increased numbers of arteriolar profiles and hypoplasia to absence of portal veins. The pathological investigations along with the animal history and clinical signs indicated a hepatic encephalomyelopathy due to a congenital portosystemic shunt.