Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs,humans and wildlife in south‐east Spain

Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.     

Propofol versus thiopental: effects on peri-induction intraocular pressures in normal dogs

ObjectiveTo determine the effects of propofol or thiopental induction on intraocular pressures (IOP) in normal dogs.Study designProspective randomized experimental study.AnimalsTwenty-two random-source dogs weighing 19.5 ± 5.3 kg.MethodsDogs were randomly assigned to receive propofol 8 mg kg⁻¹ IV (group P) or thiopental 18 mg kg⁻¹ IV (group T) until loss of jaw tone. Direct arterial blood pressure, arterial blood gasses, and IOP were measured at baseline, after pre-oxygenation but before induction, before endotracheal intubation, and after intubation.ResultsThere were no significant differences between groups with regard to weight, body condition score, breed group, or baseline or before-induction IOP, arterial blood pressure, or blood gases. The baseline IOP was 12.9 mmHg. Before endotracheal intubation, IOP was significantly higher compared to baseline and before induction in dogs receiving propofol. After intubation with propofol, IOP was significantly higher compared to thiopental and was significantly higher compared to before induction. After intubation, IOP was significantly lower compared to before intubation in dogs receiving thiopental. Propofol increased IOP before intubation by 26% over the before-induction score and thiopental increased IOP by 6% at the same interval. The IOP in group P remained 24% over the before induction score whereas thiopental ultimately decreased IOP 9% below baseline after intubation. There was no significant relationship between any cardiovascular or blood gas parameter and IOP at any time. There was no significant relationship between the changes in any cardiovascular or blood gas parameter and the changes in IOP between time points.Conclusions and clinical relevancePropofol caused a significant increase in IOP compared to baseline and thiopental. Thiopental caused an insignificant increase in IOP which decreased after intubation. Propofol should be avoided when possible in induction of anesthesia in animals where a moderate increase in IOP could be harmful.     

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

Evidence of Migration and Endophytic Presence of Agrobacterium tumefaciens in Rose Plants

Agrobacterium tumefaciens was isolated from stem tumors of several rose cultivars showing that the bacterium is the causal agent of aerial galls in rose plants. No differences were observed in the characteristics of the Agrobacterium isolates from crown or aerial galls. Stem inoculation of ten rose cultivars showed that all of them were susceptible to A. tumefaciens but differences in the size of the resulting tumors were observed. The movement of A. tumefaciens in rose plants was demonstrated using two wild type strains and two antibiotic resistant mutants. Three months after inoculation, the inoculated strains were recovered in the roots, crown and below and above the inoculation site but low numbers of pathogenic Agrobacterium cells were isolated. New tumors appeared in 5% of the noninoculated wounds. A. tumefaciens was isolated from the stem at different distances from the tumor in naturally infected plants. In symptomless commercial plants, the isolation from the roots, crown and at different stem levels demonstrated the existence of systemic and latent infections in rose. Direct isolation using a nonselective and selective media with or without a previous enrichment step were efficient methods for isolating tumorigenic Agrobacterium from the different parts of rose plants.     

2n gametes inSolanum tuberosum dihaploids

Pollen stainability over two years and microsporogenesis were investigated in seven dihaploids extracted fromSolanum tuberosum tetraploids (AVRDC-1287.19, Conchita, Merrimack and W231). Two dihaploids proved to be male-sterile and male gamete abortion is likely to be post-meiotic because a regular meiosis occurred. The other dihaploids proved to be partially male-fertile and 2n pollen producers, as well. Almost all clones showed predominantly the formation of parallel spindles. Correlations estimated between the observed abnormalities in spindle orientation and the meiotic products evidenced a high correlation between parallel/fused spindles and dyads. The dihaploids were also used as parents in crosses with tetraploids. One berry with 44 seeds was obtained from dihaploid AVI 24 used as pollen parent in 4x × 2x crosses. After 2x × 4x crosses, three dihaploids including the two male-sterile were found to be 2n egg producers. The formation of 2n gametes inS. tuberosum dihaploids is discussed in relation to a direct use of dihaploids in sexual polyploidization schemes to combine useful traits in a superior genotype at 4x level.     

Bentazone Leaching in Spanish Soils

Adsorption, incubation and soil-column experiments with bentazone [3-isopropyl-1H-2,1,3-benzothiadiazin-4(3H)-one 2,2-dioxide] were carried out in ten different soils from the marches surrounding the Doñana National Park (Huelva, SW Spain). Adsorption isotherms for the different soils showed a good fit with the Freundlich equation. Bentazone was poorly adsorbed in all the soils studied, with no significant relationship between theK_f values and soil characteristics. A significant correlation was obtained between the soil organic matter content and the distribution constant values (K_d) calculated at an equilibrium concentration of 200 μg cm⁻³. The low adsorption and non-degradation of bentazone on these soils suggest that the herbicide readily percolates through soils to reach the surface and ground waters. The mobility of bentazone through three soil columns was also studied. The mass balances carried out showed that bentazone was totally eluted from the soil columns. The theoretical model applied to explain bentazone leaching under our experimental conditions seems to be suitable for soil columns with a uniform water-flow rate.     

Soil evolution over the Quaternary period in a semiarid climate (Segre river terraces,northeast Spain)

A macro- and micromorphologic study was done on the soils from a stepped sequence of seven dated fluvial terraces in the lower Segre river valley (Lleida, northeast Spain) under a present-day semiarid Mediterranean climate. The soils have evolved from the Holocene through the early Pleistocene, providing an excellent morphostratigraphic framework for evaluating time-dependent factors influencing soil formation in a dry and calcareous environment. Throughout the chronosequence, some properties are regularly age-related specially carbonatation in subsurface horizons. The carbonates occur mainly as micrite, and although microsparite and sparite also appear in the oldest soils, they are replaced by fine-grained calcite by dissolution–reprecipitation processes (micritization process), which is active at present. Some pedological paleofeatures as the presence of sparite and recarbonated argillans in oldest terrace can be interpreted as the reflex of climatic changes during the Quaternary. In spite of this climate variability, the soils display progressive and systematic patterns of carbonate accumulation: on the lowest terraces, the soils do not yet have secondary carbonates but in the beginning Late Pleistocene calcic horizons, with carbonate pendents, are developed; these pendents increase its thickness with age although pendent growth rates decreases from Late Pleistocene to Middle Pleistocene. In the middle of the Late Pleistocene, calcic horizons evolved to petrocalcic horizons, which increase its thickness in the Middle and specially in Early Pleistocene. The presence of calcic and petrocalcic horizons is the primary basis for soil classification. This criterion is applicable not only to the soils of the lower Segre river basin, but also to many soils throughout the semiarid Mediterranean region.     

Comparative assessment of food web structure and fisheries productivity of three reservoirs in Ghana

Three Ghanaian reservoirs (Tono, Bontanga and Golinga) were compared through a food web modelling approach (Ecopath with Ecosim) to assess production characteristics and food web structures. The lakes differ in size and morphology, generating specific conditions for fish growth and production. While the two top fishery target species were Sarotherodon galilaeus (L.) and Oreochromis niloticus (L.) in all reservoirs, the mean trophic level of the catch was lowest in the largest and deepest reservoir (Tono) due to higher trophic level species occupying less accessible deep “refuge” habitats. The smallest lake had the highest fish production under optimal conditions of water supply (17.1 compared with 15.5 and 10.1 t/km²/year for lakes Bontanga and Tono), but it appears to be most vulnerable under conditions of drought. For the planning and construction of adequately sized reservoirs used for fishery and irrigation purposes, the water budget (ratio of inflow and evaporation) needs to be estimated.