Expression of interferon gamma in the brain of cats with natural Borna disease virus infection

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus, which causes a non-suppurative meningoencephalomyelitis in a wide range of animals. In cats, BDV infection leads to staggering disease. In spite of a vigorous immune response the virus persists in the central nervous system (CNS) in both experimentally and naturally infected animals. Since the CNS is vulnerable to cytotoxic effects mediated via NK-cells and cytotoxic T-cells, other non-cytolytic mechanisms such as the interferon (IFN) system is favourable for viral clearance. In this study, IFN-γ expression in the brain of cats with clinical signs of staggering disease (N=12) was compared to the expression in cats with no signs of this disease (N=7) by quantitative RT-PCR. The IFN-γ expression was normalised against the expression of three reference genes (HPRT, RPS7, YWHAZ). Cats with staggering disease had significantly higher expression of IFN-γ compared to the control cats (p-value ≤ 0.001). There was no significant difference of the IFN-γ expression in BDV-positive (N=7) and -negative (N=5) cats having clinical signs of staggering disease. However, as BDV-RNA still could be detected, despite an intense IFN-γ expression, BDV needs to have mechanisms to evade this antiviral immune response of the host, to be able to persist.     

Degradation of ochratoxin a by Brevibacterium species

The ability to degrade ochratoxin A was studied in different bacteria with a well-known capacity to transform aromatic compounds. Strains belonging to Rhodococcus, Pseudomonas, and Brevibacterium genera were grown in liquid synthetic culture medium containing ochratoxin A. Brevibacterium spp. strains showed 100% degradation of ochratoxin A. Ochratoxin α was detected and identified by high-performance liquid chromatography-mass spectrometry (HPLC-MS) as a degradation product in the cell-free supernatants. The degradation of ochratoxin A is of public concern for food and environmental safety, because it could contribute to the development of new biological ochratoxin A detoxification systems in foodstuffs. In this study, the degradation of ochratoxin A by bacteria belonging to the food chain was demonstrated for the first time.     

USE OF TRANSSPLENIC INJECTION OF AGITATED SALINE AND HEPARINIZED BLOOD FOR THE ULTRASONOGRAPHIC DIAGNOSIS OF MACROSCOPIC PORTOSYSTEMIC SHUNTS IN DOGS

We describe the use of ultrasonography‐guided percutaneous splenic injection of agitated saline and heparinized blood for the diagnosis of portosystemic shunts (PSS) in 34 dogs. Agitated saline mixed with 1 ml of heparinized autologous blood was injected into the spleen of 34 sedated dogs under sonographic guidance. The transducer was then sequentially repositioned to visualize the portal vein, the caudal vena cava, and the right atrium through different acoustic windows. It was possible to differentiate between intrahepatic and extrahepatic shunts depending on the entry point of the microbubbles into the caudal vena cava. Portoazygos shunts and portocaval shunts could be differentiated based on the presence of microbubbles in the caudal vena cava and/or the right atrium. In one dog, collateral circulation due to portal hypertension was identified. In dogs with a single extrahepatic shunt, the microbubbles helped identify the shunting vessel. The technique was also used postoperatively to assess the efficacy of shunt closure. All abnormal vessels were confirmed by exploratory laparotomy or with ultrasonographic identification of the shunting vessel. Ultrasound‐guided transsplenic injection of agitated saline with heparinized blood should be considered as a valuable technique for the diagnosis of PSS; it is easy to perform, safe, and the results are easily reproducible.     

Bioactivity Improvement of <Emphasis Type="Italic">Olea europaea</Emphasis> Leaf Extract Biotransformed by <Emphasis Type="Italic">Wickerhamomyces anomalus</Emphasis> Enzymes

Olive leaves represent a quantitatively significant by-product of agroindustry. They are rich in phenols, mainly oleuropein, which can be hydrolyzed into several bioactive compounds, including hydroxytyrosol. In this study, water extract from olive leaves ‘Biancolilla’ was analyzed for polyphenol profile, DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and protective effect on differentiated Caco-2 cells. The efficacy of two enzymatic treatments in promoting the release of bioactive phenols was investigated: a) enzymatic extract from Wickerhamomyces anomalus, characterized by β-glucosidase and esterase activities; b) commercial β-glucosidase. Composition and bioactivity of the resulting extracts were compared. The results showed that the yeast-treated extract presented hydroxytyrosol content and DPPH radical scavenging activity comparable to those obtained using commercial β-glucosidase; however, it was showed the additional presence of hydroxycinnamic acids. In experiments on Caco-2 cells, the leaf extracts promoted the recovery of cell membrane barrier at different minimum effective concentrations. The high specificity of W. anomalus enzymatic extract may represent an effective tool for the release of bioactive phenols from olive by-products.     

Breeding of Persian melon: Purpose to increase volatile compounds in melon (Cucumis melo) intergroup F1 cross population

Understanding the volatile profile of melon is an ever increasingly important task for breeders for enhancing their flavour component. For this purpose, F1 progeny and parental lines were analysed using headspace-solid phase microextraction gas chromatography–mass spectrometry-based analysis. Diallel analysis showed that most of the investigated compounds were controlled via non-additive genetic effects with general combining ability to specific combining ability ratios of less than 0.4. High heterosis results were observed in alcohols such as ethanol and octanol in both rind and mesocarp samples and benzylalcohol in the rind. It was also noted that the ester-based compounds with high heterosis values in mesocarp samples differed from those with heterosis in rind samples. These observations further emphasize the non-additive control of these compounds. It can be concluded that non-additive genetic control is strongly governing the genetic profile within the studied progeny. This finding indicated that dudaim lines as a valuable genetic material enhanced the volatile profile of odourless commercial types of ameri and inodorus melon.     

Effects of Preparation Temperature on Gelation Properties and Molecular Structure of High‐Amylose Maize Starch

In this study, 3% aqueous high‐amylose maize starch (Hylon VII) dispersions were heated to temperatures of 140–165°C. The onset and rate of gel formation was observed using a small‐strain oscillation rheometer as a function of temperature from 90 to 25°C. The gel formation clearly began earlier in high‐amylose starch paste preheated at lower temperatures, but the rate of gelation was slower and the resulting gel was weaker in comparison with starch pastes preheated at higher temperatures. In addition, the structure of the final gels was studied using large deformation compression measurements. The most rigid gel structure on the basis of small and large deformation tests was obtained for high‐amylose starch gel preheated to 150–152°C, depending on the type of measurement. The rate of gelation was also fastest in that temperature range. High‐amylose gels heated to higher temperatures lost their rigidity. The molecular weight distribution of starch molecules was measured by size‐exclusion chromatography. Heating caused extensive degradation of amylopectin, which had a great effect on amylose gel formation and the final gel properties of high‐amylose maize starch. Micrographs of Hylon VII gels showed that phase separation of starch components visible in light microscopy occurred on heating to higher temperatures.     

Demographic consequences of anthropogenic influences: Florida black bears in north-central Florida

Anthropogenic habitat fragmentation poses a serious threat to conservation of large carnivores, due to their extensive movements and potential conflicts with humans. We studied the population ecology of Florida black bears (Ursus americanus floridanus) for 6 years in two study areas in north-central Florida: Ocala National Forest (ONF), a contiguous forested habitat, and an adjacent residential community of Lynne, a fragmented habitat with substantial human activities. We estimated age-specific survival and fecundity rates of bears using data from radio-collared bears, and parameterized and analyzed stage-structured matrix population models for the two study sites and also for data pooled from both sites. Annual survival rates of adult females were lower in Lynne (0.776 ± 0.074) than in ONF (0.966 ± 0.023). While cub survival rates were higher in Lynne (0.507 ± 0.135) than in ONF (0.282 ± 0.109), the rates at both sites were substantially lower than those reported for other black bear populations. Age-specific fecundities did not vary between sites. The asymptotic population growth rate for ONF was greater than one, whereas that for Lynne was less than one. Our results suggest that anthropogenic influences (primarily road density and vehicular traffic, through their effect on adult survival) can substantially affect the population dynamics of Florida black bears and other large carnivores with large home ranges. We recommend efforts such as constructing highway underpasses, which could reduce road-related mortalities, to ensure long-term persistence of Florida black bears facing threats from rapidly increasing human influences.     

Valorization of three varieties of grape

In the present investigation, seed methanolic extracts of three Vitis vinifera (Muscat d’Italie, Syrah and Carignan) were assayed for their antioxidant activities. Results showed that there are strong variations in the contents of total phenols (440.97–121.94 mg GAE g⁻¹ DW), flavonoids (48.07–16.81 mg EC g⁻¹ DW) and tanins (37.15–14.9 mg EC g⁻¹ DW) from the studied seeds. The phenolic composition of these extracts was determined by RP–HPLC after acid hydrolysis. The main phenolic compound was quercetin with 27.2% in Muscat d’Italie, 48.8% in Syrah and 28.4% in Carignan. Besides, all seed extracts showed remarkable DPPH radical scavenging activity with IC₅₀ values ranged from 1.8 to 30 μg ml⁻¹. EC₅₀ values of reducing power activity ranged from 100 to 120 μg ml⁻¹. The high phenolic content and the considerable antioxidant activities of vitis seed extracts could potentially be considered as an expensive source of natural antioxidants.     

Genetic variability of Appaloosa horses: a study of a closed breeding population from Argentina

The genetic diversity and structure of 72 Appaloosa horses belonging to a closed breeding population from an ecological reserve in Buenos Aires, Argentina, was investigated using eight microsatellite markers from the International Society for Animal Genetics panel. Our data showed that this Appaloosa horse population had an elevated degree of genetic diversity (He= 0.746) and did not present a significant increase of homozygous individuals (F_IS~0). However, the short tandem repeats, AHT5, ASB2, HTG10 and VHL20, were not in Hardy–Weinberg equilibrium (P-value<0.05). Genetic relationships between this population and other well known horse breeds showed that Appaloosa horses from Argentina could have had their origin in the horses of the Nez Perce’s people in Idaho while other Appaloosa horses may have had influences from Andalusian and Lusitano breeds. This closed breeding population conserves an important degree of Appaloosa genetic diversity and notwithstanding its particular breeding characteristics, represents a valuable genetic resource for conservation.