Proteomic profiling of sea bass muscle by two-dimensional gel electrophoresis and tandem mass spectrometry

In this study, the proteome profile of European sea bass (Dicentrarchus labrax) muscle was analyzed using two-dimensional electrophoresis (2-DE) and tandem mass spectrometry with the aim of providing a more detailed characterization of its specific protein expression profile. A highly populated and well-resolved 2-DE map of the sea bass muscle tissue was generated, and the corresponding protein identity was provided for a total of 49 abundant protein spots. Upon Ingenuity Pathway Analysis, the proteins mapped in the sea bass muscle profile were mostly related to glycolysis and to the muscle myofibril structure, together with other biological activities crucial to fish muscle metabolism and contraction, and therefore to fish locomotor performance. The data presented in this work provide important and novel information on the sea bass muscle tissue-specific protein expression, which can be useful for future studies aimed to improve seafood traceability, food safety/risk management and authentication analysis. This work is also important for understanding the proteome map of the sea bass toward establishing the animal as a potential model for muscular studies.     

Regeneration of peroxiredoxins by p53-regulated sestrins, homologs of bacterial AhpD

Acting as a signal, hydrogen peroxide circumvents antioxidant defense by overoxidizing peroxiredoxins (Prxs), the enzymes that metabolize peroxides. We show that sestrins, a family of proteins whose expression is modulated by p53, are required for regeneration of Prxs containing Cys-SO(2)H, thus reestablishing the antioxidant firewall. Sestrins contain a predicted redox-active domain homologous to AhpD, the enzyme catalyzing the reduction of a bacterial Prx, AhpC. Purified Hi95 (sestrin 2) protein supports adenosine triphosphate-dependent reduction of overoxidized PrxI in vitro, indicating that unlike AhpD, which is a disulfide reductase, sestrins are cysteine sulfinyl reductases. As modulators of peroxide signaling and antioxidant defense, sestrins constitute potential therapeutic targets.     

Alien species in aquatic environments: a selective comparison of coastal and inland waters in tropical and temperate latitudes

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Enhancing rotifer Brachionus calyciflorus population growth with commercial probiotics

The effect of a commercial probiotic (NanoCrusta, Altacrusta, Mexico City, Mexico) on the growth performance of Brachionus calyciflorus Pallas, 1766, was evaluated. In a first approach, probiotics were supplied in four densities (2.0 × 10³, 1.1 × 10⁵, 2.1 × 10⁵ and 2.1 × 10⁶ cells/ml), alone and in combination with Chlorella vulgaris (1 × 10⁶ cells/ml). The test rotifer did not grow on the probiotic alone. However, when probiotics + C. vulgaris were added, the maximum densities (D_max; ind/ml) and population growth rates (r) observed were higher. In the second experiment, probiotics were supplied at five higher densities (2.1 × 10⁶, 4.2 × 10⁶, 8.5 × 10⁶, 1.7 × 10⁷ and 3.4 × 10⁷ cells/ml) with C. vulgaris and a control treatment with only C. vulgaris (probiotic‐free). Treatments supplied with probiotics between 2.1 × 10⁶ and 1.7 × 10⁷ cells/ml showed significantly higher D_max and r than the control treatment. The results showed a positive effect of probiotic bacteria when supplied with C. vulgaris. The best outcome showed a D_max 2.16 times and an r 1.63 times higher than the density of the control treatment. Growth rates were higher in the treatments with probiotics compared to the control. We conclude that application of NanoCrusta is feasible to improve B. calyciflorus production, but the effects need to be tested in larger scales.     

Human Impacts on Forest Biodiversity in Protected Walnut-Fruit Forests in Kyrgyzstan

We used a spatially explicit model of forest dynamics, supported by empirical field data and socioeconomic data, to examine the impacts of human disturbances on a protected forest landscape in Kyrgyzstan. Local use of 27 fruit and nut species was recorded and modeled. Results indicated that in the presence of fuelwood cutting with or without grazing, species of high socioeconomic importance such as Juglans regia, Malus spp., and Armeniaca vulgaris were largely eliminated from the landscape after 50–150 yr. In the absence of disturbance or in the presence of grazing only, decline of these species occurred at a much lower rate, owing to competitive interactions between tree species. This suggests that the current intensity of fuelwood harvesting is not sustainable. Conversely, current grazing intensities were found to have relatively little impact on forest structure and composition, and could potentially play a positive role in supporting regeneration of tree species. These results indicate that both positive and negative impacts on biodiversity can arise from human populations living within a protected area. Potentially, these could be reconciled through the development of participatory approaches to conservation management within this reserve, to ensure the maintenance of its high conservation value while meeting human needs.     

Enhancement of alkalinity tolerance in two cucumber genotypes inoculated with an arbuscular mycorrhizal biofertilizer containing Glomus intraradices

The aim of the present study was to determine whether arbuscular mycorrhizal (AM) inoculation with a biofertilizer containing clays as granular carriers, leek root pieces and Glomus intraradices spores could improve alkalinity tolerance of two cucumber genotypes, and to study the changes induced by AM at agronomical and physiological level. A greenhouse experiment was carried out to determine yield, growth, fruit quality, net photosynthesis (A_CO2), electrolyte leakage, and mineral composition of two cucumber (Cucumis sativus L.) genotypes (hybrid “Ekron” or open-pollinated variety “Marketmore”) with inoculated and noninoculated arbuscular mycorrhizal biofertilizer. Plants were supplied with nutrient solutions at two pH values (6.0 or 8.1). The high pH nutrient solution had the same basic composition with an additional 10 mM NaHCO₃ and 0.5 g l⁻¹ CaCO₃. The percentage root colonization was higher in “Marketmore” (21.8%) than “Ekron” (12.7%). Total and marketable yield and total biomass were significantly higher by 189%, 213%, and 77%, respectively, with Ekron in comparison to those recorded with Marketmore. The highest crop performance with Ekron in comparison to Marketmore was due to the improved nutritional status (higher N, P, K, Ca, Mg, Fe, Mn, and B), higher leaf area, and net photosynthesis. Increasing the concentration of NaHCO₃ from 0 to 10 mM in the nutrient solution significantly decreased yield, plant growth, A_CO2, N, P, Fe, Cu, Zn, Mn, and B concentration in leaf tissue, whereas the electrolyte leakage increased. The inoculated plants under alkaline conditions had higher total, marketable yield, and total biomass than noninoculated plant. Mycorrhizal cucumber plants grown under alkaline conditions had a higher macronutrient concentration in leaf tissue compared to noninoculated plants. The highest yield and biomass production in inoculated plants seems to be related to the capacity of maintaining higher net A_CO2 and to a better nutritional status (high P, K, Mg, Fe, Zn, and Mn and low Na accumulation) in response to bicarbonate stress with respect to −AM plants.     

Response of common wheat varieties to organic and conventional production systems across Italian locations,and implications for selection

Specific breeding for organic systems may help reduce their yield gap relative to conventional systems by exploiting genotype × system (GS) interaction. Likewise, specific breeding for distinct subregions within a region could capitalize on genotype × location (GL) interaction. Grain yield and test weight of common wheat varieties were evaluated under organic and conventional systems in ten locations spanning from northern to southern Italy, with the objectives of: (i) comparing production systems; (ii) investigating the extent of GS and GL interactions and their relationship with genotypic and environmental characteristics; and (iii) preliminarily comparing, in terms of predicted selection gains, different strategies to cope with GS and GL effects. These effects were investigated in the 2-year Data set 1 including seven genotypes. GS effects were also assessed in the annual Data sets 2 and 3 including 13 and 11 genotypes, respectively. The yield reduction of organic systems relative to conventional ones averaged 28% in Data set 1, 29% in Data set 2 and 14% in Data set 3. Organic systems also tended to a modest test weight reduction. Genetic correlations between systems ranged from high to very high (0.88 ≤ r_g ≤ 0.98) for yield and test weight, owing to nil or limited GS interaction. Broad-sense heritability tended to be higher in conventional systems than organic ones for yield in two data sets (mainly due to lower experimental error) while being similar in the two systems in the other cases. Predicted selection gains suggested nil (yield) or very modest (test weight) advantage of direct selection in organic systems relative to indirect selection in conventional systems, when targeting organic systems. The scope for selection only in conventional systems was reinforced when comparing predicted gains for selection scenarios which target both systems in relation to their foreseeable marketing importance. GL effects for yield and test weight were significant and were modeled by additive main effects and multiplicative interaction analysis. Site classification based on GL effects for yield revealed a larger subregion A including northern and central Italy and a smaller subregion B comprising southern Italy, accordingly with previous, independent studies. Yield selection only in subregion A (with indirect selection gain for subregion B) implied slightly higher predicted gain for A (+4%) and much lower gain for B (−24%) relative to independent, direct selection in each subregion. Selection for specific geoclimatic subregions may have greater importance than selection for specific production systems.     

Fine‐scale analysis of heavily invaded Italian freshwater fish assemblages

Inland waters are highly vulnerable to the introduction and spread of non‐native species, due to heavy human use of aquatic ecosystems and the natural linkages among streams and lakes. This is particularly noticeable in freshwater fish communities. To better evaluate how these communities are affected by non‐native species introductions, we conducted a fine‐scale analysis of the changes in Italian freshwater fish assemblages after species introduction. For this analysis, we collected information on fish species present in 44 basins. The present Italian freshwater fish fauna is composed of 48 native and 41 established introduced species, while a further 15 introduced species have been reported but are not yet considered naturalized. The changes in the fish assemblages mostly took place in the past 2 centuries and have increased recently, with nearly 60% of the species introduced in the past 3 decades. The number of species introduced per basin ranged from 0 to 35 (mean 10.85 ± 7.77 species/basin), and in 10 basins the number of species introduced is now equal to or even higher than the number of native species. In the past, introduced species mainly originated from America, but over the past three decades, an increase of introductions from other parts of Europe and Asia has been recorded. Our results show that basins already rich in native species tend to become even richer as a consequence of the establishment of introduced species. This confirms the trend toward a biotic homogenization of ecosystems even at a local scale, due to an increase in the human‐mediated spread of generalist species.     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

Doxycycline treatment for <Emphasis Type="Italic">Dirofilaria immitis</Emphasis> in dogs: impact on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Enterococcus</Emphasis> antimicrobial resistance

Doxycycline is an antibiotic that, in addition to the classic antibacterial use, is also prescribed to fight parasitic diseases, like heartworm disease in dogs. Despite the concern that the overuse of this antibiotic may decrease susceptibility of clinically important bacteria, the consequences of the prolonged doxycycline therapy in heartworm-infected dogs have never been studied before. We have analyzed the impact of this therapy on Staphylococcus aureus and Enterococcus antimicrobial resistance. In this study, 17 heartworm-infected dogs (10 that had completed the doxycycline treatment and 7 dogs that had not yet begun) were included. Twenty-four isolates of Staphylococcus aureus were obtained from two locations of each dog. After treatment, 73.3% of isolates were resistant to at least one antibiotic but only 22.2% of isolates before treatment. Most of doxycycline resistant isolates were obtained from dogs that have received treatment. Erythromycin resistance or intermediate susceptibility was detected in 45.6% of isolates, most of them from dogs after treatment. For Enterococci, 48 isolates were obtained from fecal samples (25 before treatment and 23 after treatment). Before treatment, 32% of isolates were resistant at least to one antibiotic while after, this data increase up to 65%. Comparing isolates before and after treatment, a clear increase in resistance to doxycycline (12% against 21.74%) and erythromycin (20% against 39.13%) was observed. Although the present work is a preliminary research, the results encourages the development of further studies to determinate the effect of prolonged doxycycline therapy on antimicrobial resistance.