Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Effect of increasing levels of seven tree species extracts added to a high concentrate diet on in vitro rumen gas output

This study was conducted to investigate the effects of increasing levels of extracts of Byrsonima crassifolia, Celtis pallida, Enterolobium cyclocarpum, Fraxinus excelsior, Ficus trigonata, Phoradendrom brevifolium and Prunus domestica on in vitro gas production (GP) and ruminal fermentation of a high concentrate diet. Plant extracts were prepared at 1 g dry matter (DM)/8 mL of solvent mixture (methanol : ethanol : water, 1:1:8) and added at levels of 0, 0.6, 1.2 and 1.8 mL/g DM of a high concentrate diet. In vitro GP was recorded at 2, 4, 6, 8, 10, 12, 24, 48 and 72 h of incubation. Increasing addition of extracts linearly increased (P < 0.001), the GP₂₄, GP₄₈ and GP₇₂ (mL/g DM), and linearly decreased (P < 0.001), the discrete GP lag time. Moreover, increasing extract doses linearly increased (P < 0.001) the asymptotic GP and decreased (P < 0.001) the rate of GP. GP₆ was not impacted by treatments and GP₁₂ increased linearly (P = 0.01) with increasing addition of extracts. Rumen pH declined linearly (P < 0.05) with increasing doses of extracts added. As no interactions (P > 0.05) occurred between the extracts and doses, it could be conclude that all extracts positively modified rumen fermentation at doses of 1.2 to 1.8 mL extract/g diet DM.     

Induced spawning of hatchery-raised Brazilian catfish, cachara Pseudoplatystoma fasciatum (Linnaeus, 1766)

In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with central nuclei and 17.5% with peripheral nuclei). Nine females in first maturation received two doses of carp pituitary extract (CPE), 0.5 mg/kg and 5.0 mg/kg; seven received two doses of human chorionic gonadotropin (hCG), 5 and 10 IU/g; five received doses of 0.5 CPE mg/kg and 5 hCG IU/g (CPE+hCG); and four received 0.9% saline (saline). Nine females from CPE and seven from hCG presented oocytes with the same diameter at the moment of oocyte release (100% with germinal vesicle breakdown and fertilization rate of 53.44±18.3 and 54.81±11.8%; larvae number of 165,330±94.1 and 158,570±20.6, respectively). The five females from CPE+hCG did not respond to the hormonal treatment. The four females from the saline group did not ovulate. In January 2002, 6 of 15 selected females that were going through the second reproductive cycle received CPE (five received hCG and four received saline), showing oocyte diameters similar to the ones in the first maturation. At stripping, CPE females had an oocyte diameter of 1062.5 μm (the hCG females had oocyte diameters ranging from 937.5 to 1125.0 μm; fertilization rates of 56.08±30.9 and 81.90±17.3%; 364,547±244 and 633,129±190, larvae, respectively). The fertilization rates and larvae number were higher in the second gonad maturation, both for CPE and hCG.     

Polyunsaturated Aldehydes from Large Phytoplankton of the Atlantic Ocean Surface (42°N to 33°S)

Polyunsaturated aldehydes (PUAs) are organic compounds mainly produced by diatoms, after cell wounding. These compounds are increasingly reported as teratogenic for species of grazers and deleterious for phytoplanktonic species, but there is still scarce information regarding concentration ranges and the composition of PUAs in the open ocean. In this study, we analyzed the spatial distribution and the type of aldehydes produced by the large-sized (>10 μm) phytoplankton in the Atlantic Ocean surface. Analyses were conducted on PUAs released after mechanical disruption of the phytoplankton cells, referred to here as potential PUAs (pPUAs). Results show the ubiquitous presence of pPUA in the open ocean, including upwelling areas, as well as oligotrophic gyres. Total pPUA concentrations ranged from zero to 4.18 pmol from cells in 1 L. Identified PUAs were heptadienal, octadienal and decadienal, with heptadienal being the most common (79% of total stations). PUA amount and composition across the Atlantic Ocean was mainly related to the nitrogen:phosphorus ratio, suggesting nutrient-driven mechanisms of PUA production. Extending the range of trophic conditions considered by adding data reported for productive coastal waters, we found a pattern of PUA variation in relation to trophic status.     

Metabolomic Tools for Secondary Metabolite Discovery from Marine Microbial Symbionts

Marine invertebrate-associated symbiotic bacteria produce a plethora of novel secondary metabolites which may be structurally unique with interesting pharmacological properties. Selection of strains usually relies on literature searching, genetic screening and bioactivity results, often without considering the chemical novelty and abundance of secondary metabolites being produced by the microorganism until the time-consuming bioassay-guided isolation stages. To fast track the selection process, metabolomic tools were used to aid strain selection by investigating differences in the chemical profiles of 77 bacterial extracts isolated from cold water marine invertebrates from Orkney, Scotland using liquid chromatography-high resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Following mass spectrometric analysis and dereplication using an Excel macro developed in-house, principal component analysis (PCA) was employed to differentiate the bacterial strains based on their chemical profiles. NMR ¹H and correlation spectroscopy (COSY) were also employed to obtain a chemical fingerprint of each bacterial strain and to confirm the presence of functional groups and spin systems. These results were then combined with taxonomic identification and bioassay screening data to identify three bacterial strains, namely Bacillus sp. 4117, Rhodococcus sp. ZS402 and Vibrio splendidus strain LGP32, to prioritize for scale-up based on their chemically interesting secondary metabolomes, established through dereplication and interesting bioactivities, determined from bioassay screening.     

Differential Adaptation of Complete and Substituted Triticale

Adaptation of seven complete and twelve substituted triticales to specific soil types has been studied, based on a series of twenty trials carried out in 1989 and 1990 across Spain. The nature of the GE interaction for grain yield was revealed by means of the additive main effects multiplicative interaction (AMMI) model and using the soil pH at the different sites as linear covariate. The percentage of the variability explained by the first principal component axis of the AMMI model was 72 and 65 % for the two years, suggesting a specific pattern of adaptation. Soil pH was the single most important environmental factor to explain the adaptation of complete and substituted types. Complete triticales outyielded substituted genotypes in the majority of sites. Triticale adaptation to acid and alkaline soils seems to be largely controlled by the single wheat/rye chromosome 2D(2R) substitution, for which both types differ. Complete triticales seem better adapted to the acid soils, whereas substituted types are, in general, more suited to alkaline soils.     

Agronomical and morphological differentiation among winter and spring triticales

A collection of 299 secondary hexaploid triticale cultivars and advanced breeding lines from 18 countries, which were considered a representative sample of the existing diversity, was evaluated for morphological and agronomical characters with autumn planting at Lleida, Spain, from 1988 to 1991. The entries were classified as having winter (84) or spring (215) growth habit and among this latter group were complete (73) or substituted (147) types according to CIMMYT's terminology. Winter and spring triticales were grouped by cluster and principal component analyses. Winter triticales were taller with longer growth cycles, longer spikes, and more spikelets per spike than spring types. At early stages they also had prostrate growth. Spring-substituted types were separated from complete material. As a group, spring-substituted triticales differed more from winter types than the spring complete genotypes, which showed intermediate characteristics. Complete types of spring habit had tendency to be taller, with longer spikes, more spikelets per spike and bigger and heavier grains than substituted triticales. Greater variation in morphological and agronomical parameters was detected among winter triticales followed by the complete-spring group.     

Study of nickel sorption onto biological sludges. possibilities for heavy metals removal treatments

Nickel adsorption onto a sludge wasting from a biological aeration tank is described in this work. This procedure is planned as an alternative method for metal removal from industrial effluents containing heavy metals in low concentrations. Experimental results of equilibrium adsorption and laboratory equipment operation are reported. Equilibrium data of nickel adsorption were fitted to the Freundlich equation. On the basis of this equation, the effect of several parameters such us metal concentration, sludge concentration and metal dosing rate were tested. Nickel removal efficiency is about 80% in the most favourable cases. Aeration during metal-sludge contact was also tested but little enhancing in metal uptake was observed. This analysis can be used to define the best operation conditions in practical cases either for metal or organic matter removal.