Administration of Probiotic Strain to Improve Sea Bream Wellness during Development

Two bacterial strains Lactobacillus fructivorans (AS17B), isolated from adult sea bream (Sparus aurata) gut, and Lactobacillus plantarum (906), isolated from human faeces, were administered contemporaneously, during sea bream development using Brachionus plicatilis and/or Artemia salina and dry feed as vectors. The probiotic treatment influenced gut colonization: at 35 days post-hatching (p.h.) L. fructivorans was not present in the gut, but the treatment induced colonization by L. plantarum. At 66 days p.h., L. fructivorans was evident also in the control; moreover, when suitable environmental conditions appeared in the post-metamorphosis gastro-intestinal tract, competition between L. plantarum and L. fructivorans occurred. At 90 days p.h., L. plantarum was displaced by L. fructivorans that became significantly higher with respect to the control. In treated groups, probiotic administration significantly decreased larvae and fry mortality.     

Definition of priority areas for forest conservation through the ordered weighted averaging method

The general objective of this study was to evaluate the ordered weighted averaging (OWA) method, integrated to a geographic information systems (GIS), in the definition of priority areas for forest conservation in a Brazilian river basin, aiming at to increase the regional biodiversity. We demonstrated how one could obtain a range of alternatives by applying OWA, including the one obtained by the weighted linear combination method and, also the use of the analytic hierarchy process (AHP) to structure the decision problem and to assign the importance to each criterion. The criteria considered important to this study were: proximity to forest patches; proximity among forest patches with larger core area; proximity to surface water; distance from roads; distance from urban areas; and vulnerability to erosion. OWA requires two sets of criteria weights: the weights of relative criterion importance and the order weights. Thus, Participatory Technique was used to define the criteria set and the criterion importance (based in AHP). In order to obtain the second set of weights we considered the influence of each criterion, as well as the importance of each one, on this decision-making process. The sensitivity analysis indicated coherence among the criterion importance weights, the order weights, and the solution. According to this analysis, only the proximity to surface water criterion is not important to identify priority areas for forest conservation. Finally, we can highlight that the OWA method is flexible, easy to be implemented and, mainly, it facilitates a better understanding of the alternative land-use suitability patterns.     

Pharmacokinetic-pharmacodynamic integration of danofloxacin after intravenous, intramuscular and subcutaneous administration to rabbits

The pharmacokinetics of danofloxacin was studied following intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) administration of 6 mg/kg to healthy rabbits. Danofloxacin concentration were determined by high-performance liquid chromatography assay with fluorescence detection. Minimal inhibitory concentrations (MICs) assay of danofloxacin against 30 strains of Staphylococcus aureus from several European countries was performed in order to compute pharmacodynamic surrogate markers. The danofloxacin plasma concentration versus time data after i.v. administration could best be described by a two-compartment open model. The disposition of i.m. and subcutaneously administered danofloxacin was best described by a one-compartment model. The terminal half-life for i.v., i.m. and s.c. routes was 4.88, 6.70 and 8.20 h, respectively. Clearance value after i.v. dosing was 0.76 L/kg.h. After i.m. administration, the absolute bioavailability was mean (+/-SD) 102.34 +/- 5.17% and the Cmax was 1.87 mg/L. After s.c. administration, the absolute bioavailability was mean (+/-SD) 96.44 +/- 5.95% and the Cmax was 1.79 mg/L. Danofloxacin shows a favourable pharmacokinetics profile in rabbits reflected by parameters such as a long half-life and a high bioavailability. However, in consideration of the low AUC/MIC indices obtained, its use by i.m. and s.c. route against the S. aureus strains assayed in this study cannot be recommended given the risk for selection of first mutant subpopulations.     

An immunohistochemical study of the tonsils in pigs with acute African swine fever virus infection

An immunohistochemical study of the tonsils was carried out to gain further insight in the pathogenesis of acute African swine fever (ASF). Twenty-one pigs were inoculated by intramuscular route with a highly virulent isolate of ASF virus and painlessly killed at 1-7dpi. Viral antigen was highly distributed in the tonsil from 3 to 4dpi and an increase in the number of monocyte-macrophages was very evident at the same days post inoculation. This phenomenon was observed together with an increase of the expression of proinflammatory cytokines (Tumour necrosis factor alpha and Interleukin-1 alpha) and the apoptosis of lymphocytes studied by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling (TUNEL) technique and haemorrhages. With these results, we can conclude that the tonsil is suffering similar lesions than those observed in other lymphoid organs in acute African swine fever, even when the route of inoculation is the intramuscular and not oral-nasal.     

Leishmania-specific isotype levels and their relationship with specific cell-mediated immunity parameters in canine leishmaniasis

In the current retrospective study, Leishmania infantum-specific IgG, IgA and IgM levels were determined by ELISA in 106 untreated dogs with clinically-patent leishmaniasis (Sx) and in 171 clinically healthy dogs (Asx) from Spain to investigate the relationship between these Ig isotypes and clinical status. In addition, we studied if different Leishmania-specific humoral pattern exists between Asx dogs with and without cellular mediated immunity (CMI). Fifty-six dogs were assessed by means of lymphoproliferation assay (LPA), interferon production (IFN) and leishmanin skin test (LST), 71 dogs by means of both LPA and IFN and 44 only by LST. Both Sx and Asx dogs produced Leishmania-specific IgG, IgA and IgM antibodies, however the levels and proportion of positive dogs for each Ig isotype were significantly higher in Sx than in Asx ones (P<0.001). Analysis of immunological profiles determined for each cellular technique (positive and negative cellular response for each technique combined with positive or negative specific humoral response) showed that Asx dogs constituted a high heterogeneous group. No correlations were observed between CMI tests and specific IgG or IgM levels. However, a significant inverse correlation was demonstrated between specific IgA levels and LPA response (Spearman's r=-0.220; P=0.035). In general, the low correlation detected between CMI tests and isotype levels might indicate that the immune response is not strongly polarized in the majority of Asx dogs. Additionally, this study suggests that dogs developing T-cell response are probably able to avoid the dissemination of the parasite at least to mucosal surfaces and, as a consequence, to produce low or background specific IgA levels. Further studies are needed to investigate the relationship between specific IgA and parasite load, especially at mucosal site.     

Sunflower mutant containing high levels of palmitic acid in high oleic background

A new sunflower mutant, CAS-12, was obtained, which has both high palmitic (≈30%) and high oleic acid contents, and also a substantial amount of palmitoleic acid (≈7%). The mutant was selected after X-ray irradiation of dry seeds of the inbred line BSD-2-423, which had normal palmitic (≈3%) and high oleic (≈88%) acid levels. The increase of palmitic and palmitoleic acids occurred at the expense of the oleic acid content, which decreased to around 55% in respect to the original line. Linoleic acid content is always under 5%. Palmitic and palmitoleic acid levels were similar to those of the high palmitic mutant CAS-5 obtained in a previous programme from a low oleic line isogenic to BSD-2-423 using a similar mutagenic treatment. In that previous programme we also selected three high stearic acid mutants using chemical mutagenic treatment on the same sunflower line (RDF-1-532). We attempted to obtain mutants in other lines but were unsuccessful. The isolation of similar mutants in isogenic parental lines illustrates the importance of the genetic background in the development of specific mutants with an altered seed oil fatty acid composition. The oil of this mutant will increase the range of potential uses of sunflower oil. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding two base populations of perennial ryegrass

Summary A program designed to develop two base populations of perennial ryegrass was initiated in 1985 in Galicia. Multitrait mass selection by a base index was practised in two genetic resources groups of wild Galician populations. Plants selected were intercrossed and the two resulting base populations (F and G) were evaluated in two locations at the spaced plant level and in plot trials with the aim of estimating the genetic parameters and the possibilities for future breeding. The two populations did not show significant differences from commercial cultivars. Production characters had low heritabilities (0.01–0.32) in both locations. Seasonal yields had medium to high family heritabilities (0.36–0.73). For the economic traits chosen, the expected genetic advances were important in both populations for summer and autumn growth in Mabegondo and spring and summer-autumn dry matter yield (1^st and 3^rd year) in Puebla de Brollón. Expected genetic advance for early spring growth was 0.62 in population F. In the autumn of 1991 equal numbers of plants, selected at each location by combined family and mass selection from each of these populations, were planted in two polycrosses with the aim of improving the genetic basis and adaptation of these populations.     

Factorial regression analysis of genotype by environment interaction in ryegrass populations, using both isozyme and climatic data as covariates

A sample of 58 natural ryegrass populations (Lolium perenne L.) from the French and Spanish oceanic coast was evaluated for three seasonal growth traits (i.e. spring, autumn and winter vigour traits) in 5 evaluation sites (three in France and two in Spain). This sample showed a high level of genotype (population) × environment (evaluation sites) interactions for the three agronomic traits. A factor regression analysis, using both isozyme frequencies of populations and climatic factors of evaluation sites as covariates, was carried out on a sub-sample of 30 populations in order to explain these interactions. This method succeeded in explaining most population × location interactions by the product of two covariates. For instance, for spring vigour trait, 72.8% of the interaction term could be explained by the use of two covariates: the PGI2-20 allelic frequency of populations and the minimum temperature of the coldest month of evaluation sites. This study shows the advantage of such a method for plant breeders who need to identify promising ryegrass populations for their breeding objectives. A number of genetic and evolutionary hypotheses are also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.