Conjugated linoleic acid and black currant seed oil in the treatment of canine atopic dermatitis: a preliminary report

Although conjugated linoleic acid (CLA) shows inhibitory effects on histamine release, eicosanoid production and pruritus in laboratory rodents, its use in canine atopic dermatitis (AD) has not been reported. The aims of this study were to assess the efficacy of CLA, black currant seed oil (BSO) or a combination of both, compared to placebo, in dogs with AD and to evaluate any changes in fatty acid metabolism with these treatments. Twenty-four dogs with AD were randomly allocated to four groups, and were treated orally each day for two months with either 1 mL/10 kg CLA (80% purity), 1 mL/10 kg pure BSO, 1 mL/10 kg CLA+1 mL/10 kg BSO, or 1 mL/10 kg sugar syrup (placebo). Serum was obtained on days 0, 30 and 60 for analysis of CLA metabolites, linoleic acid (LA), gamma-linolenic acid (GLA), dihomo-gamma-linolenic acid (DGLA) and arachidonic acid (AA). At the same time point, the owners were asked to assess pruritus and the veterinarians evaluated any skin lesions present. Although the best clinical results occurred with BSO treatment alone, improvement of clinical signs and pruritus was not significant with any of the treatments. Serum levels of GLA and DGLA significantly increased in BSO-treated dogs, but not in the CLA+BSO group. CLA at the dosage used in this study was not efficacious in treating canine AD, whereas BSO may help some dogs with AD, although further studies are necessary before this can be recommended as a treatment.     

A non-Mendelian inheritance of resistance to strobilurin fungicides in Ustilago maydis

Mutants of Ustilago maydis (DC) Corda with high resistance to azoxystrobin (RF 164 to 4714, based on EC50 values), an inhibitor of mitochondrial electron transport at the cytochrome bc1 complex, were isolated in a mutation frequency of 2.3 x 10(-7) after nitrosoguanidine mutagenesis and selection on media containing 1 microgram ml-1 azoxystrobin in addition to 0.5 mM salicylhydroxamate (SHAM), a specific inhibitor of cyanide-resistant (alternative) respiration. Oxygen uptake in whole cells was strongly inhibited in the wild-type strains by azoxystrobin (1.5 micrograms ml-1) in addition to SHAM (1 mM), but not in the mutant isolates. Genetic analysis with nine such mutant isolates resulted in progeny phenotypes which did not follow Mendelian segregation, but satisfied the criteria of non-Mendelian (cytoplasmic) heredity. In crosses between three mutant isolates with the compatible wild-type strains, the sensitivity was inherited by progeny maternally from the wild-type parent strain (criterion of uniparental inheritance). In crosses between wild-type strains and remaining mutant isolates, a continuous distribution of sensitivity in the progeny was found (criterion of vegetative segregation). The third criterion of cytoplasmic resistance (criterion of intracellular selection) was fulfilled by experiments on the stability of resistance phenotypes. With two exceptions, a reduction of resistance was observed in the mutant strains when they were grown on inhibitor-free medium. Recovery of the high resistance level was observed after they were returned to the selection medium. Cross-resistance studies with other fungicides, which also inhibit electron transport through complex III of respiratory chain, showed that mutations for resistance to azoxystrobin were also responsible for reduced sensitivity to kresoxim-methyl (RF 18 to 1199) and to antimycin-A (RF 20 to 305), which act at the Qo and Qi sites of the cytochrome bc1 complex, respectively. Studies of the fitness of azoxystrobin-resistant isolates showed that these mutations appeared to be pleiotropic, having significant adverse effects on growth in liquid culture and pathogenicity on young corn plants.     

Insights into population genetics,connectivity and demographic history of the longnosed skate Dipturus oxyrinchus (Linnaeus, 1758) in the western Mediterranean Sea

1. Genetic information is crucial for the conservation of Dipturus oxyrinchus (Linnaeus, 1758), a threatened large skate with declining populations over most of its geographical range. The main aim of the present study was to investigate the genetic structure, connectivity and demographic history of the longnosed skate in Sardinia (western Mediterranean Sea).
2. Patterns of population structure were assessed in 175 specimens from six sampling sites. Variation in two mitochondrial genes (cytochrome c oxidase subunit I (COI) and control region) highlighted high genetic diversity and low but significant genetic differentiation among sites, which clustered into three groups corresponding to the north‐west, north‐east and south Sardinian coasts.
3. The observed genetic structuring could presumably depend on a combination of past geological events, contemporary restrictions to dispersal and biological characteristics of the species (e.g. site‐fidelity, no pelagic larval stage, limited dispersal of juveniles and/or adults).
4. Demographic analyses showed signs of past population expansion, but substantial current stability of Sardinian populations. From a conservation perspective, these results are encouraging, and indicate that Sardinian populations are still large and stable, and seem not to have suffered negative side‐effects from the ever‐growing fishing pressure in the region.
5. The occurrence of genetic structuring strongly supported the close monitoring of populations to identify any erosion of their gene pool, and high genetic variability of the Sardinian D. oxyrinchus populations could thus represent priority populations for conservation purposes, providing potential sources for recolonization in cases of local extinctions in other areas of the distribution range of the species.
6. When the sequences from Sardinia were compared with those available from other areas, the data seem to exclude the possibility that the Atlantic and Mediterranean host totally isolated populations or even different species, as recently suggested. However, additional markers and a larger sampling sites are needed to confirm these findings.

     

Conservation management for Orthoptera in the Dadia reserve, Greece

The diversity patterns, the ecological structure and the typical species of the orthopteran assemblage in the Dadia reserve are investigated. The reserve was designed to protect the black vulture (Aegypius monachus) and other raptors. A total of 39 orthopteran species were found, including Paranocarodes chopardi, a pamphagid species with very restricted distribution. All species can be represented in a network of six complementary habitats, including open oak woodlands, agricultural fields separated with hedges, humid grasslands, as well as serpentine grasslands. The buffer zone of the reserve is far more important for Orthoptera conservation than the core areas, which host most of the black vulture nests. Management focusing on raptors is in general compatible with conservation of Orthoptera. We suggest the maintenance of forest openings in the buffer zone, the maintenance of forest heterogeneity, the enhancement of periodical livestock grazing, and the use of nine indicator species and Paranocarodes chopardi in the reserve monitoring program.     

Phosphate adsorption by dark alkaline vertisols in Greece

Phosphorus adsorption isotherms are presented for samples from agricultural areas in central Greece. The soils contain montmorillonitic clays, have high exchange capacities and range in pH from 6.8 to 8.1. The amounts of P adsorption are high for surface and subsurface samples and are comparable to the adsorption observed in acid kaolinitic soils. The fit of the Freundlich equation to the experimental data was significantly improved when NaHCO₃ extractable P was used as an estimate of the labile soil P to account for P already adsorbed in the soil. No conclusive relationship was established between the Freundlich parameters and selected soil properties though in some cases a values (capacity term) increased with increasing organic carbon content.     

Rotenone and rotenoids in cubè resins, formulations, and residues on olives

Rotenone and rotenoids (deguelin, beta-rotenolone (12a beta-hydroxyrotenone), tephrosin (12a beta-hydroxydeguelin), 12a alpha-hydroxyrotenone, and dehydrorotenone) were determined in cubè resins and formulations. Cubè resins from Lonchocarpus contain large quantities of deguelin (ca. 21.2%) and smaller quantities of tephrosin (ca. 3.5%) and beta-rotenolone (ca. 3.0%). The composition of commercial formulations may present very different rotenoid contents depending on the extracts used to prepare them. Because these rotenoids also present insecticide activity, the efficacy of these formulations may be very different. The storage stability and photodegradation of some rotenone formulations were studied. Rotenone and rotenoids are very sensitive to solar radiation, which degrades them rapidly, with half-lives in the order of a few tens of minutes. Some formulations show greater disappearance rates than that of cubè resin, indicating that not much attention has been paid to protecting the active ingredients from photodegradation in the formulation. A study on the residues on olives was also carried out to assess not only the rotenone content, but also that of the main rotenoids. At harvest, the residues of deguelin, tephrosin, and beta-rotenolone were 0.10, 0.06, and 0.10 mg/kg, respectively, very similar to rotenone (0.08 mg/kg), and though a few data indicate similar acute toxicity values for deguelin, only rotenone is taken into consideration in the legal determination of the residue.     

UV-C irradiation induces defence responses and improves vase-life of cut gerbera flowers

UV-C (λ = 254 nm) irradiation was effective in reducing Botrytis cinerea floret specking (i.e., lesion development) and maintaining a better postharvest quality of cut gerbera flowers. A range of UV-C doses (0.5-10.0 kJ m⁻²) was tested on ‘Ice cream’ and ‘Ecco’ gerbera flowers to activate germicidal and inducible defence mechanisms. Irradiation of B. cinerea cultures with 0.5, 1.0, 2.5 and 5.0 kJ m⁻² UV-C resulted in up to a 10-fold reduction of conidial germination percentages and significant (P < 0.05) delay of mycelium growth, compared to the non-irradiated control cultures. Moreover, lesion diameters on gerbera florets inoculated with UV-C irradiated B. cinerea cultures were reduced by up to 70%, suggesting that UV-C had a negative effect on the pathogenic strength of the fungi. Lesion diameters on florets of UV-C irradiated gerberas were reduced by up to 55% giving evidence that defence responses in the host tissue were induced. Concentration of total phenolics seemed to be unaffected by 0.5 kJ m⁻² UV-C treatment in both cultivars, but polyphenol oxidase (PPO) activity increased and remained higher compared to the non-irradiated control flowers throughout the 48 h storage period at 20 °C. The increase of PPO suggests that this enzyme might play an important role in host defence mechanisms that suppressed B. cinerea floret specking. Gerbera flowers irradiated with 1.0 or 10.0 kJ m⁻² UV-C showed improvement in vase-life by 1.8 and 2.4 d, decrease in stem break percentages by 43 and 29% and delay in stem break incidence by 3.3 and 1.3 d, respectively.     

Molecular typing of Brucella melitensis endemic strains and differentiation from the vaccine strain Rev-1

In the present study forty-four Greek endemic strains of Br. melitensis and three reference strains were genotyped by Multi locus Variable Number Tandem Repeat (ML-VNTR) analysis based on an eight-base pair tandem repeat sequence that was revealed in eight loci of Br. melitensis genome. The forty-four strains were discriminated from the vaccine strain Rev-1 by Restriction Fragment Length Polymorphism (RFLP) and Denaturant Gradient Gel Electrophoresis (DGGE). The ML-VNTR analysis revealed that endemic, reference and vaccine strains are genetically closely related, while most of the loci tested (1, 2, 4, 5 and 7) are highly polymorphic with Hunter-Gaston Genetic Diversity Index (HGDI) values in the range of 0.939 to 0.775. Analysis of ML-VNTRs loci stability through in vitro passages proved that loci 1 and 5 are non stable. Therefore, vaccine strain can be discriminated from endemic strains by allele’s clusters of loci 2, 4, 6 and 7. RFLP and DGGE were also employed to analyse omp2 gene and reveled different patterns among Rev-1 and endemic strains. In RFLP, Rev-1 revealed three fragments (282, 238 and 44 bp), while endemic strains two fragments (238 and 44 bp). As for DGGE, the electrophoretic mobility of Rev-1 is different from the endemic strains due to heterologous binding of DNA chains of omp2a and omp2b gene. Overall, our data show clearly that it is feasible to genotype endemic strains of Br. melitensis and differentiate them from vaccine strain Rev-1 with ML-VNTR, RFLP and DGGE techniques. These tools can be used for conventional investigations in brucellosis outbreaks.     

Influence of postharvest hot water treatment on nutritional and functional properties of kumquat (Fortunella japonica Lour. Swingle Cv. Ovale) fruit

The present study investigated the influence of a hot water dip (HWD) for 2 min at 50 degrees C, a standard and effective treatment for postharvest decay control of citrus fruit, on the nutritional and health-related properties of kumquats. The results show that most of the parameters examined, including titratable acidity, soluble solids content, maturity index, glucose, fructose, sucrose, ascorbic acid, dehydroascorbic acid, alpha- and gamma-tocopherols, beta-carotene, zeaxantin, rhoifolin, and antioxidant activity, were not significantly affected by treatment. The levels of beta-cryptoxanthin, narirutin, and total flavonoids increased after HWD, whereas lutein and total phenols decreased. The concentration of the essential oil and the relative percentage of the individual components of the essential oil were not affected by HWD except for the minor compound p-menta-1,5-dien-1-ol, which increased after HWD. After storage, lower levels of glucose, total sugars, beta-carotene, beta-cryptoxanthin and lutein were recorded in HWD fruit. A decrease in antioxidant activity and increases in alpha-tocopherol and total vitamin E were found both in control and HWD fruit. The influence of HWD at 50 degrees C for 2 min on individual nutraceuticals and health-related properties was thus generally low and may depend on storage conditions.