A new bacterial dehydrogenase oxidizing the lignin model compound guaiacylglycerol beta-O-4-guaiacyl ether

A lignin model compound, named in short guaiagylglycerol beta-guaiacyl ether (GGE), contains the beta-0-4 ether linkage that is common in the chemical structure of lignin. A Pseudomonas sp. (GU5) had been isolated as an organism able to grow with GGE as the sole source of carbon and energy. When grown on vanillate, the bacteria contained a NAD+ -dependent dehydrogenase converting GGE to a 355 nm absorbing product. The enzyme, named GGE-dehydrogenase, was purified about 160-fold using gel permeation, ion exchange on DEAE-Sephadex, and dye-ligand affinity chromatography. The new protein was about 52 kDa in apparent size with but one polypeptide chain after denaturation and reduction. According to several criteria, the product of GGE oxidation (Km = 12 microM) was identified as the corresponding conjugated ketone at the alpha-carbon of the C3 side-chain. The secondary alcohol function in GGE was apparently the sole target of the enzyme action. However the conversion of GGE into ketone catalyzed by the enzyme was only partial, and did not exceed 50%, probably because only one of the alpha-enantiomers was susceptible to enzyme attack. In contrast the ketone, either made by organic synthesis or by enzymic oxidation of GGE, could be totally reduced back to GGE (Km = 13 microM at pH 8.4, 8 microM at neutral pH), with NADH as the reductant, as confirmed by UV absorption and NMR spectra. Other model compounds with no primary alcoholic function, ether linkage or phenolic group were also substrates for the enzyme, confirming the specificity of GGE-dehydrogenase for the alpha-carbon position. Conjugation of the alpha-ketone with an adjacent phenolic nucleus interfered strongly with equilibrium constants and redox potentials of the system according to pH, and the enzyme displayed widely different optima with pH over 9 when oxidizing GGE, below 7 when reducing the ketone. Equilibrium studies showed that the ketone/GGE potential was -0.37 volt at pH 8.7, -0.23 volt at pH 7 (30 degrees C). The significance of this new dehydrogenase and its properties are discussed, especially in the general concern of lignin biodegradation.     

Expression of genes that encode cellular oxidant/antioxidant systems are affected by heat stress

Molecular Biology Reports - Heat stress causes critical molecular dysfunction that affects productivity in chickens. Thus, the purpose of this study was to evaluate the effect of heat stress (HS)...     

Rearing performances and environmental assessment of sea cage farming in Tunisia using life cycle assessment (LCA) combined with PCA and HCPC

Purpose

The present study aims to understand the influence of rearing practices and the contributions of production phases of fish farming to their environmental impacts and determine which practices and technical characteristics can best improve the farms’ environmental performance. Another objective is to identify the influence of variability in farming practices on the environmental performances of sea cage aquaculture farms of sea bass and sea bream in Tunisia by using principal component analysis (PCA) and hierarchical clustering on principal components (HCPC) methods and then combining the classification with life cycle assessment (LCA).

Methods

The approach consisted of three major steps: (i) of the 24 aquaculture farms in Tunisia, 18 were selected which follow intensive rearing practices in sea cages of European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) and then a typology was developed to classify the studied farms into rearing practice groups using HCPC; (ii) LCA was performed on each aquaculture farm and (iii) mean impacts and contributions of production phases were calculated for each group of farms. Impact categories included acidification, eutrophication, global warming, land occupation, total cumulative energy demand and net primary production use.

Results and discussion

Results revealed high correlation between rearing practices and impacts. The feed-conversion ratio (FCR), water column depth under the cages and cage size had the greatest influence on impact intensity. Rearing practices and fish feed were the greatest contributors to the impacts studied due to the production of fish meal and oil and the low efficiency of feed use, which generated large amounts of nitrogen and phosphorus emissions. It is necessary to optimise the diet formulation and to follow better feeding strategies to lower the FCR and improve farm performance. Water column depth greatly influenced the farms’ environmental performance due to the increase in waste dispersion at deeper depths, while shallow depths resulted in accumulation of organic matter and degradation of water quality. Cage size influences environmental performances of aquaculture farms. Thus, from an environmental viewpoint, decision makers should grant licences for farms in deeper water with larger cages and encourage them to improve their FCRs.

Conclusions

This study is the first attempt to combine the HCPC method and the LCA framework to study the environmental performance of aquacultural activity. The typology developed captures the variability among farms because it considers several farm characteristics in the classification. The LCA demonstrated that technical parameters in need of improvement are related to the technical expertise of farm managers and workers and to the location of the farm.

     

The LPS O-Antigen in Photosynthetic Bradyrhizobium Strains Is Dispensable for the Establishment of a Successful Symbiosis with Aeschynomene Legumes

The photosynthetic bradyrhizobia are able to use a Nod-factor independent process to induce nitrogen-fixing nodules on some semi-aquatic Aeschynomene species. These bacteria display a unique LPS O-antigen composed of a new sugar, the bradyrhizose that is regarded as a key symbiotic factor due to its non-immunogenic character. In this study, to check this hypothesis, we isolated mutants affected in the O-antigen synthesis by screening a transposon mutant library of the ORS285 strain for clones altered in colony morphology. Over the 10,000 mutants screened, five were selected and found to be mutated in two genes, rfaL, encoding for a putative O-antigen ligase and gdh encoding for a putative dTDP-glucose 4,6-dehydratase. Biochemical analysis confirmed that the LPS of these mutants completely lack the O-antigen region. However, no effect of the mutations could be detected on the symbiotic properties of the mutants indicating that the O-antigen region of photosynthetic Bradyrhizobium strains is not required for the establishment of symbiosis with Aeschynomene.     

Variability of <Emphasis Type="Italic">Artemia salina</Emphasis> cysts from Sabkhet El Adhibet (southeast Tunisia) with special regard to their use in aquaculture

Brine shrimp Artemia (Crustacea, Anostraca) diverge in biometry and nutritional quality. These differences in Artemia characteristics are significant not only from strain to strain but also from one harvest to another within same strain. The main objective of this study was to compare Artemia salina cysts harvested from Sabkhet El Adhibet (southeast Tunisia) on different dates between 2002 and 2007 with special regard to their use in aquaculture, using cysts and naupliar biometrics, protein, carbohydrate, and lipid content. Fatty acid profiles as well as hatching characterisation were also evaluated. Hydrated cysts measures ranged between 258.1 and 263.7 μm, while the freshly hatched nauplii of Artemia measures ranged between 458.1 and 476.1 μm. Lipid contents of the samples ranged from 16.2 to 18.3% of the dry weight. Fatty acid profiles showed that cysts from Sabkhet El Adhibet contain a high quantity of eicosapentaenoic acid (20: 5n-3) with a percentage ranging between 7.8 and 14.3% of the dry weight. The highest hatching efficiency was obtained for decapsulated cysts collected in 2007 (139500 nauplii g⁻¹ of cysts). Cysts treated with hydrogen peroxide had a hatching percentage of 14.49 to 42.99%. The hatching synchronization time for untreated cysts varied between 23.5 to 27.4 h.     

Longitudinal random effects models for genetic analysis of binary data with application to mastitis in dairy cattle

A Bayesian analysis of longitudinal mastitis records obtained in the course of lactation was undertaken. Data were 3341 test-day binary records from 329 first lactation Holstein cows scored for mastitis at 14 and 30 days of lactation and every 30 days thereafter. First, the conditional probability of a sequence for a given cow was the product of the probabilities at each test-day. The probability of infection at time t for a cow was a normal integral, with its argument being a function of "fixed" and "random" effects and of time. Models for the latent normal variable included effects of: (1) year-month of test + a five-parameter linear regression function ("fixed", within age-season of calving) + genetic value of the cow + environmental effect peculiar to all records of the same cow + residual. (2) As in (1), but with five parameter random genetic regressions for each cow. (3) A hierarchical structure, where each of three parameters of the regression function for each cow followed a mixed effects linear model. Model 1 posterior mean of heritability was 0.05. Model 2 heritabilities were: 0.27, 0.05, 0.03 and 0.07 at days 14, 60, 120 and 305, respectively. Model 3 heritabilities were 0.57, 0.16, 0.06 and 0.18 at days 14, 60, 120 and 305, respectively. Bayes factors were: 0.011 (Model 1/Model 2), 0.017 (Model 1/Model 3) and 1.535 (Model 2/Model 3). The probability of mastitis for an "average" cow, using Model 2, was: 0.06, 0.05, 0.06 and 0.07 at days 14, 60, 120 and 305, respectively. Relaxing the conditional independence assumption via an autoregressive process (Model 2) improved the results slightly.     

The In Vitro and In Vivo Inhibitory Effects of Some Sulfonamide Derivatives on Rainbow Trout (Oncorhynchus Mykiss) Erythrocyte Carbonic Anhydrase Activity

The in vitro and in vivo inhibitory effects of 5-(3α, 12α-dihydroxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (1), 5-(3α, 7α, 12α-trihydroxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (2), 5-(3α, 7α, 12α-triacetoxy-5-β-cholanamido)-1,3,4-thiadiazole-2-sulfonamide (3) and acetazolamide on rainbow trout (Oncorhynchus mykiss) (RT) erythrocyte carbonic anhydrase (CA) were investigated. The RT erythrocyte CA was obtained by affinity chromatography with a yield of 20.9%, a specific activity of 422.5?EU/mg protein and a purification of 222.4-fold. The purity of the enzyme was confirmed by SDS-PAGE. Inhibitory effects of the sulfonamides and acetazolamide on the RT erythrocyte CA were determined using the CO₂-Hydratase method in vitro and in vivo studies. From in vitro studies, it was found that all the compounds inhibited CA. The obtained I₅₀ value for the sulfonamides (1), (2) and (3) and acetazolamide were 0.83, 0.049, 0.82 and 0.052?μM, respectively. From in vivo studies, it was observed that CA was inhibited by the sulfonamides (1), (2) and (3) and acetazolamide.