Protein changes as robust signatures of fish chronic stress: a proteomics approach to fish welfare research

Background Aquaculture is a fast-growing industry and therefore welfare and environmental impact have become of utmost importance. Preventing stress associated to common aquaculture practices and optimizing the fish stress response by quantification of the stress level, are important steps towards the improvement of welfare standards. Stress is characterized by a cascade of physiological responses that, in-turn, induce further changes at the whole-animal level. These can either increase fitness or impair welfare. Nevertheless, monitorization of this dynamic process has, up until now, relied on indicators that are only a snapshot of the stress level experienced. Promising technological tools, such as proteomics, allow an unbiased approach for the discovery of potential biomarkers for stress monitoring. Within this scope, using Gilthead seabream (Sparus aurata) as a model, three chronic stress conditions, namely overcrowding, handling and hypoxia, were employed to evaluate the potential of the fish protein-based adaptations as reliable signatures of chronic stress, in contrast with the commonly used hormonal and metabolic indicators. Results A broad spectrum of biological variation regarding cortisol and glucose levels was observed, the values of which rose higher in net-handled fish. In this sense, a potential pattern of stressor-specificity was clear, as the level of response varied markedly between a persistent (crowding) and a repetitive stressor (handling). Gel-based proteomics analysis of the plasma proteome also revealed that net-handled fish had the highest number of differential proteins, compared to the other trials. Mass spectrometric analysis, followed by gene ontology enrichment and protein-protein interaction analyses, characterized those as humoral components of the innate immune system and key elements of the response to stimulus. Conclusions Overall, this study represents the first screening of more reliable signatures of physiological adaptation to chronic stress in fish, allowing the future development of novel biomarker models to monitor fish welfare.This study received Portuguese national funds from FCT - Foundation for Science and Technology through project UIDB/04326/2020 and project WELFISH (Refª 16–02-05-FMP-12, “Establishment of Welfare Biomarkers in farmed fish using a proteomics approach”) financed by Mar2020, in the framework of the program Portugal 2020. Cláudia Raposo de Magalhães acknowledges an FCT PhD scholarship, Refª SFRH/BD/138884/2018. Denise Schrama acknowledges an FCT PhD scholarship, Refª SFRH/BD/136319/2018.info:eu-repo/semantics/publishedVersio

Fish processing and digestion affect parvalbumins detectability in Gilthead Seabream and European seabass

Consumption of aquatic food, including fish, accounts for 17% of animal protein intake. However, fish consumption might also result in several side-effects such as sneezing, swelling and anaphylaxis in sensitized consumers. Fish allergy is an immune reaction to allergenic proteins in the fish muscle, for instance parvalbumin (PV), considered the major fish allergen. In this study, we characterize PV in two economically important fish species for southern European aquaculture, namely gilthead seabream and European seabass, to understand its stability during in vitro digestion and fish processing. This information is crucial for future studies on the allergenicity of processed fish products. PVs were extracted from fish muscles, identified by mass spectrometry (MS), and detected by sandwich enzyme-linked immunosorbent assay (ELISA) after simulated digestion and various food processing treatments. Secondary structures were determined by circular dichroism (CD) after purification by anion exchange and gel filtration chromatography. In both species, PVs presented as α-helical and β-sheet structures, at room temperature, were shown to unfold at boiling temperatures. In European seabass, PV detectability decreased during the simulated digestion and after 240 min (intestinal phase) no detection was observed, while steaming showed a decrease (p < 0.05) in PVs detectability in comparison to raw muscle samples, for both species. Additionally, freezing (−20 °C) for up to 12 months continued to reduce the detectability of PV in tested processing techniques. We concluded that PVs from both species are susceptible to digestion and processing techniques such as steaming and freezing. Our study obtained preliminary results for further research on the allergenic potential of PV after digestion and processing

Early-to-mid idiopathic Parkinson’s disease shows a more cytotoxic but declined CD8-regulatory peripheral immune profile

Parkinson’s disease (PD) is the second most common neurodegenerative disease. Brain neuroinflammation plays a role in PD pathogenesis. However, the involvement of the peripheral immune system has not been systematically investigated. Here we analyzed >700 combinatorial immunological features in fresh blood of 28 early-to-mid-stage PD patients and 24 matched controls. We found an enhanced cytotoxic immune profile in idiopathic PD patients (iPD), with a higher frequency of terminally-differentiated effector CD8 T (TEMRA), late-differentiated CD8+ natural killer T cells and neutrophils. This immune profile was intensified by elevated serum granzyme A, reduced percentages of CD8+FOXP3+ regulatory T cells and group 2 innate lymphoid cells with immunosuppressive or tolerance-inducing functions. The frequency of CD8 TEMRA was negatively correlated with disease duration, suggesting a contribution to PD pathogenesis. Our work provides a comprehensive map on disturbed peripheral adaptive and innate immune cells in early-to-mid iPD, proposing easily-accessible candidates for early diagnosis and treatments

Immunogenicity of a Promiscuous T Cell Epitope Peptide Based Conjugate Vaccine against Benzo[a]pyrene: Redirecting Antibodies to the Hapten

The prototype polycyclic aromatic hydrocarbon benzo[a]pyrene (B[a]P) is an environmental pollutant and food contaminant of epidemiological importance. To protect against adverse effects of this ubiquitous carcinogen, we developed an immunoprophylactic strategy based on a B[a]P-protein conjugate vaccine to induce B[a]P specific antibodies (Grova et al., Vaccine. 2009;27:4142–51). Here, we investigated in mice the efficacy of B[a]P-peptide conjugates based on promiscuous T cell epitopes (TCE) into further improve this approach. We showed that B[a]P-peptide conjugates induced very different levels of hapten-specific antibodies with variable functional efficacy, depending on the carrier. In some cases peptide carriers induced a more efficient antibody response against B[a]P than tetanus toxoid as a protein carrier, with the capacity to sequester more B[a]P in the blood. Reducing the carrier size to a single TCE can dramatically shift the antibody bias from the carrier to the B[a]P. Conjugates based on the TCE FIGITEL induced the best anti-hapten response and no antibodies against the carrier peptide. Some peptide conjugates increased the selectivity of the antibodies for the activated metabolite 7,8-diol-B[a]P and B[a]P by one or two orders of magnitude. The antibody efficacy was also demonstrated in their ability to sequester B[a]P in the blood and modulate its faecal excretion (15–56%). We further showed that pre-existing immunity to the carrier from which the TCE was derived did not reduce the immunogenicity of the peptide conjugate. In conclusion, we showed that a vaccination against B[a]P using promiscuous TCEs of tetanus toxin as carriers is feasible even in case of a pre-existing immunity to the toxoid and that some TCE epitopes dramatically redirect the antibody response to the hapten. Further studies to demonstrate a long-term protection of an immunoprophylactic immunisation against B[a]P are warranted

L'Ame gauloise : revue bi-mensuelle littéraire, politique et sociale / fondateur administrateur Armand Gilles ; directrice littéraire Gab

11 mars 19231923/03/11 (A9,N125)-1923/03/11

Additional file 12: of Integration of Ixodes ricinus genome sequencing with transcriptome and proteome annotation of the naïve midgut

Combined direct acyclic graphs for the different GO categories “molecular function” (A), “biological process” (B) and “cellular component” (C). (PDF 90 kb

Additional file 13: of Integration of Ixodes ricinus genome sequencing with transcriptome and proteome annotation of the naĂŻve midgut

FDR analysis of mass spectrometry search results. Mass spectrometry search results against concatenated target-decoy databases including protein scores and cumulative FDR. (XLSX 138 kb

Additional file 7: of Integration of Ixodes ricinus genome sequencing with transcriptome and proteome annotation of the naïve midgut

Distribution of annotated sequences by GO categories “molecular function” (A), “biological process” (B) and “cellular component” (C) of level 2 of the combined direct acyclic graph for the annotated transcriptome sequences. The number in brackets represent the number of sequences that are annotated to this GO term. (PDF 495 kb

Additional file 4: of Integration of Ixodes ricinus genome sequencing with transcriptome and proteome annotation of the naĂŻve midgut

Homology comparison to Acari genomes (except I. scapularis ). A description of the best match including e-value, identity, bit score and hit length of a homology comparison against all Acari genomes registered at NCBI Genbank at the time of analysis except I. scapularis, namely Dermatophagoides farinae, Metaseiulus occidentalis, Rhipicephalus microplus, Tetranychus urticae and Varroa destructor, are listed for each contig. (XLSX 1023 kb

Immunogenicity of B[a]P-peptide conjugates after tetanus toxoid pre-vaccination.

<p>(A) Endpoint titers (serum dilution reaching 5 times the background) for TT specific IgG antibodies determined by indirect ELISA for sera pre-immunised with tetanus toxoid (TT) followed by B[a]P-peptide or B[a]P-TT conjugate vaccination. (B) B[a]P specific IgG antibodies with and without pre-vaccination. Results are presented for each mouse (○) and median value (–). There was no statistical significant difference between animal with and without pre-vaccination (One way ANOVA procedure followed by Student-Newman-Keuls-t test). (C, D) Antibody selectivity determined by competitive ELISA in sera immunised with B[a]P peptide or B[a]P-TT conjugates with (○) and without (•) tetanus toxoid (TT) pre-vaccination. B[a]P (C) and 7,8-diol-B[a]P (D) were used as competitors to compete for binding of specific antibodies to B[a]P-ovalbumin as the coated antigen. The IC₅₀ (concentration of competitor for 50% inhibition) was calculated to determine the antibody specificity for each tested competitor. The IC₅₀ is inverse correlated to the antibody affinity. Results are presented for each mouse (circle) and median value (–).</p