Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

Gastrointestinal parasitism reduces the plasma availability of doramectin in lambs

A study was undertaken to investigate the effect of parasitism on plasma availability and pharmacokinetic behaviour of doramectin (DRM) in lambs. Fourteen parasitised grey face Suffolk lambs (26.9 ± 1.5 kg bodyweight) were selected for the study. Seven pairs of lambs were allocated to two groups to obtain an approximately even weight distribution. Group I (non-parasitised) was pre-treated with three repeated administrations of 5 mg/kg fenbendazole to maintain a parasite free condition. In group II (parasitised), the lambs did not receive any anthelmintic treatment. After the 85-day pre-treatment period, both groups of animals were treated with DRM by subcutaneous (SC) injection in the shoulder area at 200 μg/kg. Throughout the experimental period, both groups were maintained together under similar feeding and management conditions. Blood samples were collected by jugular venepuncture at different set times between 0.5 h and 60 days post-treatment. After plasma extraction and derivatisation, samples were analysed by high performance liquid chromatography (HPLC) with fluorescence detection. A computerised kinetic analysis was performed and the data were compared using the Student’s paired t test.The parent molecule was detected in plasma between 30 min and either day 20 (parasitised) or day 35 (non-parasitised) post-DRM treatment. The AUC values of the parasitised group (143.0 ± 18 ng d/mL) were significantly lower (P < 0.05) than those observed in the parasitically naïve animals (229.6 ± 21.7 ng d/mL). The mean residence time (MRT) in the parasitised group (3.4 ± 0.3 days) was significantly shorter (P < 0.05) than in the healthy group (6.6 ± 0.6 days). Study results have shown that parasitic disease, through alteration in the body condition, can produce significant changes in the plasma disposition of DRM when administered SC to parasitised lambs.     

Perceptions of veterinarians and clients to expressions of clinical uncertainty

OBJECTIVES: The aim of this study was to explore the attitudes of veterinarians and clients towards expressions of clinical uncertainty. METHODS: Questionnaires that assessed the respondent's attitudes towards expressions of clinical uncertainty were completed by clients at six small animal practices. In addition, questionnaires that evaluated what veterinarians thought their client's attitudes would be towards expressions of clinical uncertainty were completed by veterinarians. The responses from clients and veterinarians were statistically compared. RESULTS: Veterinarians significantly underestimated the desire of clients to be told about uncertainties in treatment and significantly overestimated how the expression of uncertainty would generally reduce client confidence. Veterinarians significantly overestimated the loss of client confidence resulting from saying "I am not sure about this" and from asking a nurse for advice. Both clients and veterinarians considered that verbal expressions of uncertainty would lead to a greater reduction in client confidence than behavioural expressions of uncertainty. CLINICAL SIGNIFICANCE: This study suggests that most clients want to be told about their veterinarian's clinical uncertainties, and how this uncertainty is expressed can markedly influence client confidence.     

Effects of live prey availability on growth and survival in the early stages of cuttlefish Sepia officinalis (Linnaeus, 1758) life cycle

This investigation examined the effects of live prey availability on growth and survival of Sepia officinalis. Two independent experiments, comprising two feed rations each, were performed, using adequate prey size. In the first experiment, cuttlefish hatchlings were fed live mysids, Paramysis nouvelli [(feed ratio I (fr I)], at 15% body weight per day (bw day^?1) (fr I₁₅) and 30% bw day^?1 (fr I₃₀). In the second experiment, juvenile cuttlefish were fed live Atlantic ditch shrimp, Paleomonetes varians (fr II), under the same experimental design. In both experiments, the final mean weight, feeding rate and instantaneous growth rate were higher when animals were fed feed ratio fr II₁₅ and fr II₃₀ (30% bw day^?1). The results indicate that prey availability influenced weight gain, irrespective of the prey used, during the first 2 months of cuttlefish life cycle. This effect seems to be more noticeable when a certain limit of prey is achieved. For cuttlefish fed fr II, the optimal prey density is thought to be under 2.5 g prey L^?1 (i.e. 14 shrimp L^?1). Results indicate that cuttlefish can withstand prey densities up to 120 mysids L^?1 for cuttlefish up to 3 weeks old and 19 shrimps L^?1 for cuttlefish up to 6 weeks old.     

Effects of binder type and binder addition on the growth of juvenile Palaemonetes varians and Palaemon elegans (Crustacea: Palaemonidae)

This investigation compared the effects of using two binders of no nutritional value, lignosol and agar, and the binding method, microbinding and microcoating, on the growth of juvenile Atlantic ditch shrimp, Palaemonetes varians, and rockpool prawn, Palaemon elegans. Shrimp were fed one basal diet which varied solely in binder type and method of inclusion (Diet A, with lignosol added by microbinding, Diet B, with agar added by microbinding, and Diet C with lignosol added by microcoating) for a period of 45 days. At the end of the experiment significant differences in the final wet weight were found between shrimps fed Diet A and Diet C and between shrimps fed Diet B and Diet C (P < 0.001), but no significant differences were found between Diet A and Diet B (P > 0.05). These results were verified for both species. Significant differences were found between the linear regression slopes of the three tested diets for P. varians (F _3,411 = 3.41; P < 0.04), and for P. elegans (F _3,431 = 10.51; P < 0.0003). Palaemon elegans grew faster than P. varians and at the end of the experiment significant differences were found between the final wet weight of the two species (F _3,842 = 48.17, P < 0.0001). The growth parameters analysed, weight gain (WG), thermal-unit growth coefficient (TGC), and food conversion rate (FCR) were always higher for P. elegans. Results indicate that use of either lignosol or agar, added to the diet by microbinding, is advisable.     

Fractional extraction of compounds from grape seeds by supercritical fluid extraction and analysis for antimicrobial and agrochemical activities

White grape seeds were subjected to sequential supercritical fluid extraction. By increasing the polarity of the supercritical fluid using methanol as a modifier of CO(2), it was possible to fractionate the extracted compounds. Two fractions were obtained; the first, which was obtained with pure CO(2), contained mainly fatty acids, aliphatic aldehydes, and sterols. The second fraction, obtained with methanol-modified CO(2), had phenolic compounds, mainly catechin, epicatechin, and gallic acid. The fractions were bioassayed. Antimicrobial activities were checked on human pathogens, and a high degree of activity was obtained with the lipophilic fraction. Agrochemical activities on phytopathogenic fungi and activities on the etiolated wheat coleoptile bioassay were also checked. The more polar fraction was active in the latter bioassay.     

Residue level, persistence, and storage performance of citrus fruit treated with fludioxonil

The potential of postharvest dip treatments with fludioxonil (FLU) (a synthetic analogue of the bacterial metabolite of pyrrolnitrin), in controlling postharvest decay caused by Penicillium digitatum and Penicillium italicum of citrus fruit was investigated in comparison with the conventional fungicide imazalil (IMZ). The ultrastructural changes of fruit epicuticular wax was investigated as a function of water dip temperature, and the possible role of these changes was related to residue accumulation under FLU treatment. Residues retained by fruit were determined as a function of fungicide concentration, dip temperature, and fruit storage conditions. Scanning electron microscopy analysis revealed that fruit dipping in water at 30 or 40 degrees C did not cause differences in cuticular wax's ultrastructure in comparison to control fruit, while treatments at 50, 55, or 60 degrees C caused the disappearance of wax platelets, resulting in relatively homogeneous skin surface, due to partial "melting" of epicuticular wax. Residues of FLU in fruit treated at 20 or 50 degrees C were significantly correlated with the doses of fungicide applied. When equal amounts of fungicide were employed, the residue concentrations were notably higher (from 2.6- to 4-fold) in fruit treated at 50 degrees C than in fruit treated at 20 degrees C. The dissipation rate of FLU in "Salustiana" and "Tarocco" oranges was lower in fruit subjected to treatment at 50 degrees C. The minimal FLU concentration for almost complete decay control in artificially wounded fruit during 7-d storage at 20 degrees C was 400 mg/L active ingredient (ai) in fruit treated at 20 degrees C and 100 mg/L ai in fruit treated at 50 degrees C. Results on nonwounded Tarocco oranges subjected to 3 weeks of simulated quarantine conditions at 1 degrees C, plus 6 weeks of standard storage at 8 degrees C and an additional two weeks of simulated marketing period (SMP) at 20 degrees C revealed that almost complete decay control with FLU applications of 100 mg/L at 50 degrees C and 400 mg/L at 20 degrees C resulted in ca. 0.8 mg/kg FLU fruit residues, in agreement with results on wounded citrus fruit. When equal concentrations and temperatures were applied, FLU treatments were as effective as IMZ. In vitro trials showed a low sensitivity to FLU against P. digitatum and P. italicum isolates. MIC values for the complete inhibition of mycelium growth were >or=100 microg/mL, while ED(50) values ranged from 0.1 to 1 microg/mL for P. digitatum and from 1 to >100 microg/mL for P. italicum. The latter result suggests that care should be taken to avoid exclusive application of FLU in a sustainable program for management of fruit decay. However, integrating fungicide application and hot water dip may reduce the possibility of selecting fungicide-resistant populations of the pathogen, by increasing the effectiveness of the treatment.     

A New Bioassay Platform Design for the Discovery of Small Molecules with Anticancer Immunotherapeutic Activity

Immunotherapy takes advantage of the immune system to prevent, control, and eliminate neoplastic cells. The research in the field has already led to major breakthroughs to treat cancer. In this work, we describe a platform that integrates in vitro bioassays to test the immune response and direct antitumor effects for the preclinical discovery of anticancer candidates. The platform relies on the use of dendritic cells that are professional antigen-presenting cells (APC) able to activate T cells and trigger a primary adaptive immune response. The experimental procedure is based on two phenotypic assays for the selection of chemical leads by both a panel of nine tumor cell lines and growth factor-dependent immature mouse dendritic cells (D1). The positive hits are then validated by a secondary test on human monocyte-derived dendritic cells (MoDCs). The aim of this approach is the selection of potential immunotherapeutic small molecules from natural extracts or chemical libraries.