In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Effect of dietary Schizochytrium microalga oil and fish oil on plasma cholesterol level in rats

The purpose of the study was to test the hypothesis that the dietary oils with different content of n‐3 polyunsaturated fatty acids (PUFA) eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) affect plasma lipid level in rats in a different degree. The diets with 6% of fish oil (FO) and Schizochytrium microalga oil (SchO; EPA+DHA content in the diets 9.5 + 12.3 and 2.6 + 29.5% of the sum of total fatty acids, respectively) were used; the diet with 6% of safflower oil (high content of n‐6 PUFA linoleic acid, 65.5%; EPA+DHA content 0.7 + 0.9%) was used as a control. The difference between FO and SchO was established only in the case of plasma triacylglycerol (TAG) level: plasma TAG of the FO‐fed rats did not differ from the control rats (p > 0.05), while SchO decreased (p < 0.05) plasma TAG to 46% of the control. On the other hand, FO and SchO decreased (p < 0.05) total plasma cholesterol (TC) in rats in the same extent, to 73% of the control. Regarding the underlying mechanisms for the TC decrease, both SchO and FO up‐regulated hepatic Insig‐1 gene (181 and 133% of the control; p < 0.05), which tended (p = 0.15 and p = 0.19 respectively) to decrease the amount of hepatic nSREBP‐2 protein (61 and 66% of the control). However, neither SchO nor FO influenced hepatic 3‐hydroxy‐3‐methyl‐glutaryl‐CoA reductase gene expression (p > 0.05); SchO (but not FO) increased (p < 0.05) low‐density lipoprotein receptor mRNA in the liver. It was concluded that the decrease of total plasma cholesterol might be caused by an increased cholesterol uptake from plasma into the cells (in the case of SchO), but also by other (in the present study not tested) mechanisms.     

The effects of agricultural fields and human settlements on the use of rivers by wildlife in the mid-Zambezi valley,Zimbabwe

After the eradication of the Tse-Tse fly in the Mid-Zambezi valley, human settlements and fields extended mainly along the main rivers. In order to investigate the consequences of this human development on wildlife diversity we monitored three rivers of the Mid-Zambezi valley in Zimbabwe: Angwa, Manyame and Kadzi. The rivers were divided in segments of 200 m which were checked for spoors in order to assess the number of species and the number of individuals that used the segments. Human settlements were also recorded. We used a GIS to define the spatial characteristics of the fields present along the rivers, and related them to the distribution and abundance of wild species spoors in the river beds and banks. Our results show that the number of species in one segment of the river decreased with the increasing size of the field area bordering the segment. For all the major ungulate species, the numbers of individuals recorded per segment decreased with increasing field area. A similar trend was observed for small and medium-sized carnivores, though they were in lower numbers when present. Our analyses thus confirm that the extension of human agriculture in wildlife areas has an impact on most wild species, but we also define some threshold value of field size above which there seem to be an acceleration of the decrease in wildlife density and diversity: 3.2 ha for medium and small herbivores and carnivores; only the elephant seem to tolerate larger field area with a threshold value of 32 ha.This revised version was published online in May 2005 with corrections to the Cover Date.     

Testing variability in pathogenicity of Phytophthora cinnamomi

Forty eight isolates of Phytophthora cinnamomi from various host plants in France (35 isolates) and in other countries were tested for pathogenicity. Seedlings of chestnut, northern red oak, pine and eucalyptus were infected by soil contamination. Taproots, stems and bark strips of plants of chestnut and different oak species were inoculated with mycelium agar disks. Results of the different experiments were in good agreement. All isolates appeared pathogenic to all the different test species but with variable levels of virulence. Isolates with consistent low or high level of virulence, which could be used as standards in further studies, were identified. Interaction between P. cinnamomi isolates and host plant species was significant in terms of lesion lengths. These interactions could not be related to host from which P. cinnamomi was isolated. Consistent with this, in Quercus rubra, the isolate-provenance interaction was not significant. This feature is encouraging for provenance screening for resistance to P. cinnamomi in this species. The variation in virulence was not related to other isolate characteristics (mating type, electrophoretic type, age).     

Neuro-imaging the serotonin 2A receptor as a valid biomarker for canine behavioural disorders

The serotonergic system is disturbed in different mood and affective disorders, with especially the serotonin (5-HT) 2A receptor involved in impulsive aggressiveness and anxiety. The aim of the study was to evaluate the involvement of the brain 5-HT 2A receptor in dogs with different behavioural disorders. Three groups of drug naive dogs were studied: 22 dogs showing impulsive aggressive behaviour, 22 showing normal behaviour, and 22 showing anxious behaviour. The serotonin 2A receptor was evaluated with Single Photon Emission Computed Tomography (SPECT) and the serotonin 2A receptor-selective radiopharmaceutical (123)I-R91150. A serotonin 2A receptor binding index (BI), proportional to the cortical receptor density, was calculated. A receiver operating characteristic (ROC) analysis was performed to determine cut-off values at which optimal sensitivity and specificity are achieved and to evaluate the general performance of the BI in reflecting the state of the dog, i.e., impulsive aggressive, normal or anxious. Significantly (P<0.0056) altered 5-HT 2A receptor binding indices were found in bilateral frontal, temporal and occipital cortical brain areas of the dogs behaving abnormally, with consistently increased BI in impulsive aggressive dogs and decreased BI in anxious dogs. These results provide clear evidence for a disturbed serotonergic balance in canine impulsive aggression and anxiety disorders. A right frontal cut-off value of ≥1.92 with 86.4% sensitivity and 2.3% (1-specificity) was obtained for the impulsive aggressive dogs. Differentiating the anxious dogs from the rest of the population was possible with a cut-off value of ≤1.73 with 86.4% sensitivity and 18.2% (1-specificity). We conclude that SPECT imaging with the radioligand (123)I-R91150 can be a helpful tool in evaluating the involvement of the serotonin 2A receptor in the complex mechanisms of impulsive aggressive and anxious behaviour. The 5HT-2A binding index of the right frontal cortex appears to be a valid biomarker in differentiating the studied canine behavioural disorders.     

Studies on the various sugar-urea and sugar-starch-urea relationships with reference to the fermentation in the rumen and the digestibility of nutrients in fattening cattle]

Recommendations are given saying that sugar-to-crude fibre rations of not more than 1.7:1 and sugar-to-urea ratios of 12:1 should be used in the fattening of cattle fed urea-supplemented rations of roughages in which 40% to 50% of the crude protein content of the ration had been replaced by urea. The amount of sugar per kg live-weight should not exceed 4 g. Higher quantities reduce the digestibility of crude fibre and, additionally, decrease the NH3 level and the pH in the rumen. At the same time the amount of volatile fatty acids is increased while the content of ruminal acetic acid decreases. Variations in the sugar-to-starch ratio had no statistically significant influence on the nitrogen balance whereas it was found that increasing quantities of sugar significantly decreased the digestibility of crude fibre. The proportional content of sugar in the readily metabolisable carbohydrates contained in the ration should not exceed 20% to 50%.     

3 TESLA MAGNETIC RESONANCE IMAGING OF THE OCCIPITOATLANTOAXIAL REGION IN THE NORMAL HORSE

The aim of this study was to describe the appearance of the ligamentous structures of the occipitoatlantoaxial (OAA) region in the normal horse by 3 tesla (3T) magnetic resonance imaging (MRI). The MRI images of the longitudinal odontoid ligament, tectorial membrane, dorsal and ventral atlantoaxial ligaments, dorsal atlantooccipital membrane with its reinforcing ligaments, and the lateral atlantooccipital ligaments of 10 horse cadavers were evaluated. All ligaments and membranes were identified in all planes, except for the lateral atlantooccipital ligament in the sagittal plane due to its cranioventrolateral course. All were iso to mildly hypointense to musculature of the neck in T1W with the exception of the tectorial membrane that was moderately hypointense; moderately hypointense in PD‐SPIR, and markedly hypointense (isointense to cortical bone) in T2W. The PD‐SPIR was the best sequence to identify all ligaments and membranes from their cranial and caudal attachments. The longitudinal odontoid ligament, ventral atlantoaxial ligament, and reinforcing bands of the dorsal atlantooccipital membrane presented a characteristic striped heterogeneous signal behavior thought to be due to fibrocartilaginous content. The remaining ligaments and membranes showed homogeneous signal intensity. Special anatomical features in this species such as the fan‐shaped longitudinal odontoid ligament, absence of the transverse ligament and presence of the ventral atlantoaxial ligament were documented. Ligamentous structures that stabilize the equine OAA region were described with MRI in this study and these findings could serve as an anatomic reference for those cases where instability of this region is suspected.     

Subolesin/akirin orthologs from Ornithodoros spp. soft ticks: cloning, RNAi gene silencing and protective effect of the recombinant proteins

Subolesin/akirin is a well characterized protective antigen highly conserved across vector species and thus potentially useful for the development of a broad-spectrum vaccine for the control of arthropod infestations including hard ticks, mosquitoes, sand flies and the poultry red mite Dermanyssus gallinae. Soft ticks could be also targeted by this vaccine if proved that the soft tick subolesin orthologs are conserved and induce protective immune responses too. However, to date no soft tick subolesin orthologs have been fully characterized nor tested as recombinant antigens in vaccination trials. The objectives of the present work were to clone and characterize the subolesin orthologs from two important vector species of soft ticks as Ornithodoros erraticus and O. moubata, to evaluate the effect of subolesin gene silencing by RNAi, and to test the protective value of the recombinant antigens in vaccination trials. The obtained results demonstrate that both soft tick subolesins are highly conserved showing more than 69% and 74% identity with those of hard ticks in their nucleotide and amino acid sequences, respectively. Additionally, we demonstrate that both soft ticks possess fully operative RNAi machinery, and that subolesin gene silencing by dsRNA injection inhibits oviposition indicating the involvement of subolesin in tick reproduction. Finally, vaccination with the recombinant soft tick subolesins induced a partial protective effect resulting in the reduction of the oviposition rate. These preliminary results encourage further studies on the use of recombinant subolesins as vaccines for the control of soft tick infestations, either alone or in combination with other specific molecules.     

Clinical and in vitro efficacy of amoxicillin against bacteria associated with feline skin wounds and abscesses

A clinical trial involving 122 cats with infected skin wounds or abscesses presented to 10 veterinary clinics was conducted to evaluate the efficacy of 2 oral amoxicillin drug products (a paste and a suspension). A 2nd objective of the study was to identify bacteria involved in such infections and verify their in vitro sensitivity to amoxicillin. Samples of wound exudate were harvested at the time of presentation and submitted for aerobic and anaerobic culture. The sensitivity to amoxicillin of isolates thought to be infecting agents was tested, using a standard minimum inhibitory concentration method. Pasteuralla multocida and obligate anaerobes of the genera Prevotella, Fusobacterium, and Porphyromonas were the most frequently isolated pathogens. Overall, their in vitro susceptibility to amoxicillin was very good. Both drug products were clinically efficacious with a global success rate of 95.1% for cats administered oral amoxicillin at 11-22 mg/kg bodyweight (mean 13.8 mg/kg bodyweight) twice daily for 7 to 10 days.