The effect of feeding a novel multistrain yeast fraction on European seabass (Dicentrachus labrax) intestinal health and growth performance

Fish were fed a single‐strain yeast fraction (SsYF; 2 g/kg) or a multistrain yeast fraction (MsYF; 0.8 g/kg) for 10 weeks. The results demonstrated significant (p ≤ 0.03) elevations in weight gain, specific growth rate, protein efficiency ratio, and feed conversion ratio in fish fed the yeast fraction‐supplemented diets. In the distal intestine, a significant elevation in microvilli density was observed after 5 and 10 weeks of dietary supplementation with MsYF and SsYF, respectively, compared to control fed fish (p < 0.001). A significant elevation (p = 0.02) in the perimeter ratio was observed in fish fed diets supplemented with the yeast fractions. After 10 weeks of feeding on the experimental diets, Rt‐qPCR demonstrated a significant downregulation (p < 0.05) in the stress response genes, heat‐shock protein 70 (hsp70) and proliferating cell nuclear antigen (pcna), in fish fed diets supplemented with the yeast fractions. Significant (p < 0.05) elevations in interleukin 1‐beta (il1β) and interleukin‐10 (il10) gene expression were observed in fish fed diets supplemented with the MsYF compared to the other dietary groups. These findings suggest that feeding an MsYF specifically at a lower incorporation rate < 1 g/kg, compared to a commercial SsYF at 2 g/kg, is effective in improving the intestinal health status and growth performance of European seabass.     

Challenges of thermal nociceptive threshold testing in the donkey

ObjectiveTo evaluate a thermal nociceptive threshold (TNT) testing device in the donkey, and the influence of potential confounding factors on TNTs.AnimalsTwo groups (Group 1 and Group 2) of eight castrated male donkeys aged 4–9 years, weighing 105–170 kg.MethodsTNTs were measured by heating a thermal probe on skin until an end-point behaviour (threshold temperature) or a cut-out temperature (51 °C) was reached. The withers and the dorsal aspect of the distal limb were used as sites for TNT testing. The effects on TNT of different confounding factors: the limb tested; rate of heating; and ambient temperature were evaluated. Data were analyzed using general linear models, and Mann-Whitney tests, p < 0.05 was considered significant.ResultsEnd-point behaviours (skin twitch or donkey looking at test device) when the thermal probe heated the withers were observed in approximately half of tests. TNT was (mean ± SD) 46.8 ± 2.85 °C. Subsequently the limb was evaluated as the test site in Group 1 followed by Group 2 donkeys; end-point behaviour being a foot-lift. In Group 1, 72% of tests ended in an end-point behaviour but the response rate was lower in Group 2 (20%), although TNTs were similar [(47.6 ± 3.3) and (47.3 ± 3.0) °C respectively] for responding animals. Rate of heating, ambient temperature and laterality (right or left) did not affect thresholds, but mean TNT was significantly higher in the forelimb (48.5 ± 2.8 °C) than the hind limb (47.4 ± 2.8 °C) (p = 0.012).ConclusionsWhen a thermal probe cut-out temperature of 51 °C was used in TNT testing in the donkey a high proportion of tests did not produce an identifiable end point behaviour. Higher cut-out temperatures damaged the skin. Under these conditions, thermal nociceptive threshold testing appears not be an appropriate analgesiometry technique in the donkey.Clinical relevanceTNT testing under these conditions is not suitable form of analgesiometry for donkeys.     

Photoinhibition of photosynthesis in needles of two cypress (Cupressus sempervirens) clones

Photoinhibition of photosynthesis and photosynthetic recovery were studied in detached needles of cypress (Cupressus sempervirens L.) Clones 52 and 30 under controlled conditions of high irradiation (about 1900 micromol m(-2) s(-1) for 60 min; HL treatment), followed by 60 min in darkness. The degree of photoinhibition was determined based on the ratio of variable to maximum chlorophyll fluorescence (Fv/Fm), which is a measure of the potential efficiency of photosystem II (PSII), and on electron transport measurements. The Fv/Fm ratio declined in needles of both clones in response to the HL treatment. Minimal fluorescence (Fo) increased in HL-treated needles of both clones. The HL treatment decreased rates of whole-chain and PSII activity of isolated thylakoids more in Clone 52 than in Clone 30. In needles of both clones, PSI activity was less sensitive to photoinhibition than PSII activity. In the subsequent 60-min dark incubation, fast recovery was observed in needles of both clones, with PSII efficiencies reaching similar values to those in non-photoinhibited needles. The artificial exogenous electron donors diphenyl carbazide (DPC), hydroxylamine (NH2OH) and manganese chloride (MnCl2) failed to restore the HL-induced loss of PSII activity in needles of Clone 30, whereas DPC and NH2OH significantly restored PSII activity in photoinhibited needles of Clone 52. Quantification of the PSII reaction center protein D1 and the 33-kDa protein of the water-splitting complex following HL treatment of needles revealed pronounced differences between Clone 52 and Clone 30. The large decrease in PSII activity in HL-treated needles was caused by the marked loss of D1 protein and 33-kDa protein in Clone 30 and Clone 52, respectively.     

Maltoporin (LamB protein) contributes to the virulence and adhesion of Aeromonas veronii TH0426

Aeromonas veronii is one of the main pathogens causing freshwater fish sepsis and ulcer syndrome. More and more cases have shown that it has become an important zoonotic and aquatic agent. In this study, a A. veronii TH0426 mutant strain (ΔlamB) with an in‐frame deletion removed nucleotides 10–1,296 of the lamB gene was firstly constructed to investigate its functions. The results showed that the LD₅₀ value of the mutant ΔlamB to zebrafish and mice was 13.7‐fold and 5.6‐fold higher than those of the wild‐type strain, respectively. The toxicity of wild‐type strain to EPC cells was 2.1‐fold and threefold higher than those of ?lamB when infected for 1 and 2 hr. Furthermore, the ability of biofilm formation and the adhesion and invasion to EPC cells of ?lamB significantly decreased for 5.6‐fold and 1.8‐fold separately. In addition, motility detection result indicated that ?lamB lost the swimming ability. The results of flagellar staining and TEM demonstrated that the flagella of ?lamB were shed. In general, the deletion of lamB gene caused a significant decrease in the virulence and adhesion of A. veronii TH0426, and it can be known that the lamB gene of A. veronii plays a crucial role in the pathogenesis.     

Length‐based assessment of larval lamprey population structure at differing spatial scales

1. Perspectives on lamprey management contrast between pest control in the US Great Lakes and species conservation in the Pacific Northwest, New Zealand, and Europe. Five lamprey species are listed in the European Union (EU) Habitats Directive (HD) as requiring conservation measures. Assessments of HD ‘conservation status’ for these lampreys mainly target the larval (ammocoete) stage.
2. Larval lamprey populations can be assessed by evaluating presence or absence, density and demographic structure in riverine samples. Demographic structure has typically been described from length–frequency data using qualitative approaches and the statistical assignment of age classes. Length‐based indicators (LBIs) may provide a more rapid and flexible framework.
3. The demographic structure of a fish population can be described by univariate length metrics: length range (L_RANGE) and 90^th percentile of length (L₉₀). This study used a pooled data set from seven Irish catchments to estimate a reference point (RP) value for each metric corresponding to a healthy larval lamprey population. Two LBIs were then derived that can be estimated for an observed (OBS) population as LBI_RANGE = L_{RANGE_OBS}/L_{RANGE_RP} and LBI₉₀ = L_{90_OBS}/L_{90_RP}.
4. Simulated lamprey population length structures, representing a range of status values, were used to develop a reference gradient (RG) to support population assessment using LBIs. The assessment framework was applied to each of the seven Irish catchments, and also to subcatchment and sampling‐site scales within the Barrow system. The LBIs and RG suggested that most catchments had favourable conservation status overall, whereas smaller assessment scales revealed a spread of status values.
5. The proposed framework allows the rapid assessment of demographic structure and comparison across systems. There is potential to track states across sampling sites, events, and legislative reporting cycles, and to interpret change in relation to the local environment. The general approach may be adaptable for other fish species monitored during juvenile stages.

     

Sensitivity to information upscaling of agro-ecological assessments: Application to soil organic carbon management

Upscaling of agro-ecological indicators applied in regional analyses is sensitive to scale issues of the input data. This study develops a methodology to quantify this sensitivity for an indicator of soil organic carbon (SOC) dynamics at the farming system level. A reference case consists of seven fully described farms in northern Italy. Both upscaling in complexity by substituting measured input with estimated input and upscaling in space by extending the methods to farms not included in the reference case are addressed. The indicator increased with 3-107% at four farms after substituting measured management input with that estimated by an expert, whereas it remained unchanged or decreased at the other three farms. Taking the modal value from a cluster of pedological input did not lead to additional uncertainty in most cases, and only slightly increased it in others. We evaluated spatial upscaling by including 733 farms divided in 18 clusters that were described with less information as compared to the reference farms. Within each cluster, we observed relevant variability of the indicator (coefficients of variation of 12-43%), as a consequence of the heterogeneity of farms comprised in each cluster. In each cluster we calculated the indicator for one virtual farm, defined by using modal values for basic farm inputs. In this case the indicator was highly correlated (R² = 0.98) with the average of the values obtained using measured basic farm inputs. We conclude that upscaling in complexity and space introduces uncertainty in the values of the indicator compared to the reference case. The extent of such differences depends on the variability of the systems under analysis and on indicator sensitivity.     

Assay of aroma active components of virgin olive oils from southern Italian regions by SPME-GC/ion trap mass spectrometry

An SPME-GC/ion trap method was exploited to determine the chromatogram of volatile compounds of organic olive oils of southern Italian regions. The method is based on the assay of the terminal species of the "lipoxygenase pathway", which are present in the volatile fraction of the sampled compounds. Ethyl isobutanoate was used as internal standard in either the EI or CI ionization mode. The absolute concentration values of each analyte were evaluated through good-to-excellent calibration curves. Case studies on oils obtained from different cultivars or harvesting times are presented. The quantitative data for each compound were subjected to principal component analysis to characterize the different cultivars of this work.     

Use of reverse micelles for the simultaneous extraction of oil, proteins, and glucosinolates from cruciferous oilseeds

Cruciferous oilseeds are important sources of oil, proteins, and glucosinolates (GLs), potentially available when biorefinery processes are used. The proposed extraction technology is based on the use of reverse micelles (RMs) made with cetyltrimethylammonium bromide (CTAB) dispersed in organic solvent. The physicochemical properties of this extraction system and the good water solubility of many high value compounds, such as GLs and some proteins, permit the simultaneous extraction of oil, and these products from cruciferous oilseed meals. This procedure is based on three main steps: (i) seed conditioning; (ii) solid-liquid extraction by RM solution; and (iii) back-transfer of the RM solution for recovery of the extracted compounds. The method makes it possible to simultaneously extract almost the same amount of oil as with pure organic solvents used in the current extraction plants and more than 90% of soluble proteins and GLs. It is a promising biorefinery technology alternative to traditional oil extraction processes.     

Effect of improved fallow on crop productivity, soil fertility and climate-forcing gas emissions in semi-arid conditions

The impacts of fallow on soil fertility, crop production and climate-forcing gas emissions were determined in two contrasting legumes, Gliricidia sepium and Acacia colei, in comparison with traditional unamended fallow and continuous cultivation systems. After 2 years, the amount of foliar material produced did not differ between the two improved fallow species; however, grain yield was significantly elevated by 55% in the first and second cropping season after G. sepium compared with traditional fallow. By contrast, relative to the unamended fallow, a drop in grain yield was observed in the first cropping season after A. colei, followed by no improvement in the second. G. sepium had higher foliar N, K and Mg, while A. colei had lower foliar N but higher lignin and polyphenols. In the third year after fallow improvement, a simulated rainfall experiment was performed on soils to compare efflux of N₂O and CO₂. Improved fallow effects on soil nutrient composition and microbial activity were demonstrated through elevated N₂O and CO₂ efflux from soils in G. sepium fallows compared with other treatments. N₂O emissions were around six times higher from this nitrogen-fixing soil treatment, evolving 69.9 ngN₂O–N g⁻¹soil h⁻¹ after a simulated rainfall event, compared with only 8.5 and 4.8 ngN₂O–N g⁻¹soil h⁻¹ from soil under traditional fallow and continuous cultivation, respectively. The findings indicate that selection of improved fallows for short-term fertility enhancement has implications for regional N₂O emissions for dry land regions.     

Effects of pig slurry application on soils and soil humic acids

The effect of three annually consecutive additions of pig slurry at two rates (90 and 150 m3 x ha(-1) x year(-1) on soils and soil humic acids (HAs) was investigated in a field experiment under semiarid conditions. Soils and pig slurries were analyzed by standard methods. The HAs were isolated from soils and pig slurry by a conventional procedure based on alkaline extraction, acidic precipitation to pH 1, purification by repeated alkaline dissolutions and acidic precipitations, water washing, dialysis, and final freeze-drying. The HAs obtained were analyzed for elemental (C, H, N, S, and O) and acidic functional group (carboxylic and phenolic) composition, and by UV-vis, FT-IR, fluorescence, and ESR spectroscopies. With respect to the control soil, the pig slurry amended soils had greater pH and electrical conductivity, slightly larger total N content, and smaller values of C/N ratio. A decrease of total organic C was observed only in soils amended for 2 and 3 years at the higher slurry rate. With respect to control soil HA, pig slurry HA was characterized by larger contents of S- and N-containing groups, smaller acidic functional group and organic free radical contents, a prevalent aliphatic character, extended molecular heterogeneity, and smaller aromatic polycondensation and humification degrees. Amendment with pig slurry HA determines a number of modifications in soil HAs, including increase of C, S, and COOH contents, C/N ratios, and aliphaticity and decrease of extraction yields and N, O, phenolic OH, and organic free radical contents. These effects are generally more evident after the first year of slurry application and tend to disappear with increasing number of treatments. Most probably, over the years the slightly humified slurry HA is mineralized through extended microbial oxidation, whereas only the most recalcitrant components, such as S-containing, phenolic, and aliphatic structures, are partially accumulated by incorporation into soil HA.