Early and abrupt salinity reduction impacts European eel larval culture.

Reducing water salinity towards iso-osmotic conditions is a common practice applied in euryhaline fish farming to limit osmoregulation costs and enhance growth. In this respect, the present study investigated the timing of salinity reduction in an abrupt manner during European eel (Anguilla anguilla) larval culture by examining associated impacts on morphological and molecular levels. Larvae from 3 different parental combinations (families) were reared at constant 36 psu for 6 days (control) or subjected to a direct reduction to 18 psu on 1, 2, or 3 days post-hatch. Overall, salinity reduction enhanced growth and survival, resulting from more efficient energy resource utilization. In the control group, expression of growth-related igf2 remained constant, demonstrating a steady growth progression, while igf1 expression increased over time only for the salinity reduced treatments, potentially qualifying as a useful biomarker for growth performance. Even though each parental combination seems to have a different capacity to cope with salinity alterations, as observed by family-driven water-transport-related aquaporin (aqp1, aqp3) gene expression, it could be inferred that the abrupt salinity change is generally not stressful, based on non-upregulated heat shock proteins (hsp70, hsp90). However, the applied salinity reduction (irrespective of timing) induced the development of pericardial edema. As such, we conclude that despite the positive effect of salinity reduction on early growth and survival, the long-term benefit for eel larval culture lies in establishing a protocol for salinity reduction, at a precise developmental time point, without causing pericardial malformations.

Type of hormonal treatment administered to induce vitellogenesis in European eel influences biochemical composition of eggs and yolk-sac larvae.

Egg biochemical composition is among the main factors affecting offspring quality and survival during the yolk-sac stage, when larvae depend exclusively on yolk nutrients. These nutrients are primarily embedded in the developing oocytes during vitellogenesis. In aquaculture, assisted reproduction procedures may be applied enabling gamete production. For the European eel (Anguilla anguilla), reproductive treatment involves administration of pituitary extracts from carp (CPE) or salmon (SPE) to induce and sustain vitellogenesis. In the present study, we compared the influence of CPE and SPE treatments on offspring quality and composition as well as nutrient utilization during the yolk-sac stage. Thus, dry weight, proximal composition (total lipid, total protein), free amino acids, and fatty acids were assessed in eggs and larvae throughout the yolk-sac stage, where body and oil-droplet area were measured to estimate growth rate, oil-droplet utilization, and oil-droplet utilization efficiency. The results showed that CPE females spawned eggs with higher lipid and free amino acid contents. However, SPE females produced more buoyant eggs with higher fertilization rate as well as larger larvae with more energy reserves (estimated as oil-droplet area). Overall, general patterns of nutrient utilization were detected, such as the amount of total lipid and monounsaturated fatty acids decreasing from the egg stage and throughout the yolk-sac larval stage. On the contrary, essential fatty acids and free amino acids were retained. Notably, towards the end of the yolk-sac stage, the proximal composition and biometry of surviving larvae, from both treatments, were similar.

Epidemiology of panic disorder and subthreshold panic symptoms in the Greek general population.

Panic disorder (PD) is a common anxiety disorder with severe social and health consequences in the lives of individuals who suffer from it. General population studies that attempt to measure the prevalence of this disorder across the world suggest that a 1.7% to 4.7 % of adults and adolescents suffer from Panic Disorder. In Greece, research analyzing the abovementioned matters is limited, and previous studies were put forward in small samples. The aim of the present study was to describe the prevalence and sociodemographic associations of panic disorder (PD) and related subthreshold panic symptoms in the general population of Greece and to appraise the comorbidity, use of services and impact on quality of life of these syndromes. This was a secondary analysis of the 2009-2010 psychiatric morbidity survey carried out in a representative sample of the Greek general population (4894 participants living in private households, 18-70 years, response rate 54%). Psychiatric disorders were assessed with the computerized version of the revised Clinical Interview Schedule (CIS-R). Quality of life was assessed with the EuroQoL EQ-5D generic instrument. The utilization of health services was examined by making relevant questions. Finally, direct questions were used to assess sociodemographic and socioeconomic factors According to our findings, 1.87% of the participants (95% confidence interval [CI]: 1.50-2.26%) met criteria for PD and 1.61% met criteria for subclinical PD (95% CI: 1.26-1.96%). There was a clear female preponderance for both PD (p=0.001) and Sub-PD (p=0.01). In addition, 3.48% of the participants reported having experienced panic attacks during the past week (95% confidence interval [CI]: 2.98-4.01%). PD or subclinical PD was independently associated with a limited number of sociodemographic and socioeconomic variables especially after the adjusted analysis. Both panic related conditions involved significant reductions in quality of life and elevated utilization of health services for both medical and psychological reasons in comparison to healthy participants. In conclusion, PD and subclinical panic symptoms were common in the general Greek population with substantial comorbidity and impaired quality of life. The observed use of the general and psychological health services among adults with panic symptoms and its temporal and economic consequences calls for more efficient diagnostic and treatment policies.

Temperature induced variation in gene expression of thyroid hormone receptors and deiodinases of European eel (Anguilla anguilla) larvae.

Thyroid hormones (THs) are key regulators of growth, development, and metabolism in vertebrates and influence early life development of fish. TH is produced in the thyroid gland (or thyroid follicles) mainly as T4 (thyroxine), which is metabolized to T3 (3,5,3'-triiodothyronine) and T2 (3,5-diiodothyronine) by deiodinase (DIO) enzymes in peripheral tissues. The action of these hormones is mostly exerted by binding to a specific nuclear thyroid hormone receptor (THR). In this study, we i) cloned and characterized thr sequences, ii) investigated the expression pattern of the different subtypes of thrs and dios, and iii) studied how temperature affects the expression of those genes in artificially produced early life history stages of European eel (Anguilla anguilla), reared in different thermal regimes (16, 18, 20 and 22 °C) from hatch until first-feeding. We identified 2 subtypes of thr (thrα and thrß) with 2 isoforms each (thrαA, thrαB, thrßA, thrßB) and 3 subtypes of deiodinases (dio1, dio2, dio3). All thr genes identified showed high similarity to the closely related Japanese eel (Anguilla japonica). We found that all genes investigated in this study were affected by larval age (in real time or at specific developmental stages), temperature, and/or their interaction. More specifically, the warmer the temperature the earlier the expression response of a specific target gene. In real time, the expression profiles appeared very similar and only shifted with temperature. In developmental time, gene expression of all genes differed across selected developmental stages, such as at hatch, during teeth formation or at first-feeding. Thus, we demonstrate that thrs and dios show sensitivity to temperature and are involved in and during early life development of European eel.

Development of rapidly dissolving pellets within the Quality by Design approach.

The purpose of this study was the development of novel, fast disintegrating, effervescent pellets by employing the direct pelletization technique as a single step process. In line with the Quality by Design (QbD) regulatory framework, statistical experimental design was extensively applied to correlate significant formulation and process variables with the critical quality attributes of the product. Pellets were studied with regards to sphericity, size and size distribution. In contrast to the existing multiparticulate platforms, this development integrated only water-soluble excipients to facilitate the multifunctional use of the final dosage form. The application of a screening fractional factorial design augmented to a full factorial design set the roadmap for the rational selection of the composition and process parameters, revealing in parallel the positive contribution of the powder feeder on the CQAs, when the critical process and formulation factors were properly adjusted. The response surface methodology was exploited for the final process optimization phase, which allowed the construction of appropriate mathematical models connecting the input variables and the CQAs under study. The implementation of the desirability function, lead to the optimum formulation and process settings for the production of pellets with narrow size distribution and geometric mean diameter of approximately 800 µm. In conclusion, using a lean approach supported by design of experiments (DoE) techniques within the QbD framework, a novel multifunctional formulation platform has been developed.

'RETCIF': a rapid, sensitive method for detection of viruses, applicable for large numbers of clinical samples.

Rapid detection of viruses in clinical samples is important for continuing appropriate antiviral treatment and discontinuing unnecessary antibacterial treatment, as well as for excluding viral pathogens. Yet detection of viral agents may require numerous susceptible cell lines. Even with the shell vial culture method, it is cumbersome for handling large volumes of specimens. A procedure has been developed, which is time and cost-saving and uses specific cell lines in a 96-well microtitre plate and monoclonal antibodies (RETCIF-rapid enhanced tissue culture immunofluorescence). Each clinical sample was inoculated into 12 different wells with five different cell lines. Enhancement was achieved by sonication, centrifugation and hormonal supplementation to the medium used. Cytomegalovirus (CMV), herpes simplex virus (HSV) and respiratory viruses were detected by monoclonal antibodies on day 2, whilst varicella zoster virus (VZV) and enteroviruses were detected on days 5 and 7, respectively. During July-December 1998, 3298 patient specimens were compared by RETCIF and a modified shell vial method. Either or both methods isolated 779 viruses (24% positivity rate), whilst both methods detected 621. Of the 779 viruses, 87% (679) were isolated by the shell vial method in an average time of 4.9 days. For RETCIF the respective rate was 92.5% (721), in an average time of 3.0 days. The RETCIF method is a time-saving procedure, with higher isolation rates than the shell vial method.