A summer heat wave decreases the immunocompetence of the mesograzer, Idotea baltica

Extreme events associated with global change will impose increasing stress on coastal organisms. How strong biological interactions such as the host–parasite arms-race are modulated by environmental change is largely unknown. The immune system of invertebrates, in particular phagocytosis and phenoloxidase activity response are key defence mechanisms against parasites, yet they may be sensitive to environmental perturbations. We here simulated an extreme event that mimicked the European heat wave in 2003 to investigate the effect of environmental change on the immunocompetence of the mesograzer Idotea baltica. Unlike earlier studies, our experiment aimed at simulation of the natural situation as closely as possible by using long acclimation, a slow increase in temperature and a natural community setting including the animals’ providence with natural food sources (Zostera marina and Fucus vesiculosus). Our results demonstrate that a simulated heat wave results in decreased immunocompetence of the mesograzer Idotea baltica, in particular a drop of phagocytosis by 50%. This suggests that global change has the potential to significantly affect host–parasite interactions

Transcriptome characterization of the South African abalone Haliotis midae using sequencing-by-synthesis

<p>Abstract</p> <p>Background</p> <p>Worldwide, the genus <it>Haliotis </it>is represented by 56 extant species and several of these are commercially cultured. Among the six abalone species found in South Africa, <it>Haliotis midae </it>is the only aquacultured species. Despite its economic importance, genomic sequence resources for <it>H. midae</it>, and for abalone in general, are still scarce. Next generation sequencing technologies provide a fast and efficient tool to generate large sequence collections that can be used to characterize the transcriptome and identify expressed genes associated with economically important traits like growth and disease resistance.</p> <p>Results</p> <p>More than 25 million short reads generated by the Illumina Genome Analyzer were <it>de novo </it>assembled in 22,761 contigs with an average size of 260 bp. With a stringent <it>E</it>-value threshold of 10^-10, 3,841 contigs (16.8%) had a BLAST homologous match against the Genbank non-redundant (NR) protein database. Most of these sequences were annotated using the gene ontology (GO) and eukaryotic orthologous groups of proteins (KOG) databases and assigned to various functional categories. According to annotation results, many gene families involved in immune response were identified. Thousands of simple sequence repeats (SSR) and single nucleotide polymorphisms (SNP) were detected. Setting stringent parameters to ensure a high probability of amplification, 420 primer pairs in 181 contigs containing SSR loci were designed.</p> <p>Conclusion</p> <p>This data represents the most comprehensive genomic resource for the South African abalone <it>H. midae </it>to date. The amount of assembled sequences demonstrated the utility of the Illumina sequencing technology in the transcriptome characterization of a non-model species. It allowed the development of several markers and the identification of promising candidate genes for future studies on population and functional genomics in <it>H. midae </it>and in other abalone species.</p

Hemolymph microbiome of Pacific oysters in response to temperature, temperature stress and infection

Microbiota provide their hosts with a range of beneficial services, including defense from external pathogens. However, host-associated microbial communities themselves can act as a source of opportunistic pathogens depending on the environment. Marine poikilotherms and their microbiota are strongly influenced by temperature, but experimental studies exploring how temperature affects the interactions between both parties are rare. To assess the effects of temperature, temperature stress and infection on diversity, composition and dynamics of the hemolymph microbiota of Pacific oysters (Crassostrea gigas), we conducted an experiment in a fully-crossed, three-factorial design, in which the temperature acclimated oysters (8 or 22 °C) were exposed to temperature stress and to experimental challenge with a virulent Vibrio sp. Strain. We monitored oyster survival and repeatedly collected hemolymph of dead and alive animals to determine the microbiome composition by 16s rRNA gene amplicon pyrosequencing. We found that the microbial dynamics and composition of communities in healthy animals (including infection survivors) were significantly affected by temperature and temperature stress, but not by infection. The response was mediated by changes in the incidence and abundance of operational taxonomic units (OTUs) and accompanied by little change at higher taxonomic levels, indicating dynamic stability of the hemolymph microbiome. Dead and moribund oysters, on the contrary, displayed signs of community structure disruption, characterized by very low diversity and proliferation of few OTUs. We can therefore link short-term responses of host-associated microbial communities to abiotic and biotic factors and assess the potential feedback between microbiota dynamics and host survival during disease

Characterisation of shrimp haemocytes and plasma components by monoclonal antibodies

Characterisation of shrimp haemocytes and plasma components by monoclonal antibodie

Quelles stratégies alternatives aux antibiotiques en aquaculture ?

Malgré des réglementations contraignantes, l’usage des antibiotiques en préventif est encore répandu en aquaculture, particulièrement pendant les phases critiques (stades précoces, métamorphose, transferts d’animaux), mais aussi chez des animaux en croissance. En plus des améliorations que l’on peut encore apporter en matière de zootechnie et de prophylaxie traditionnelle, des méthodes alternatives sont maintenant disponibles ou en développement. Les préparations microbiennes commercialisées pour les élevages terrestres sont de plus en plus utilisées pour les élevages de crevettes et de poissons, mais chaque espèce ou élevage demanderait des expérimentations particulières pour déterminer les produits et les doses les plus efficaces. En effet, les réponses des animaux à l’ajout des probiotiques peuvent être variables et l’absence de données fiables freine leur application en routine. Les probiotiques d’origine terrestre contenant des Lactobacillus, des Bacillus ou d’autres bactéries de genres connexes, ou bien encore des levures, ne conviennent pas pour les mollusques bivalves comme les huîtres, les coquilles St Jacques, les palourdes. Seules quelques bactéries marines sélectionnées protègent les larves de bivalves contre les infections bactériennes. Cependant, l’utilisation pratique de ces bactéries gram (-) pose de nombreux problèmes d’autorisation légale, de production, de conservation et de distribution. L’intérêt des probiotiques réside dans leurs effets multiples, qui associent à des activités antibactériennes, des effets sur l’hôte telles que la stimulation de la réponse immunitaire ou celle de la croissance, bien que les mécanismes d’action ne soient pas clairement identifiés. Par contre ils n’ont pas la même efficacité que les antibiotiques pour stopper une infection. Les prébiotiques comme les fructo-oligosaccharides constituent une autre possibilité d’améliorer la santé des animaux, et des essais ont montré leur efficacité chez les alevins de turbot par exemple. Une troisième alternative est représentée par les peptides antimicrobiens. Ces molécules ont un large spectre d’activité antimicrobienne. Ils peuvent tuer des bactéries gram (-) et gram (+), des champignons ou des virus enveloppés. Plusieurs de ces peptides viennent d’être découverts chez les invertébrés marins (crevettes et huîtres), où ils sont un élément essentiel de la défense de ces animaux sans immunité acquise. Ils pourraient remplacer avantageusement les antibiotiques à terme. Ils devraient générer moins de résistances chez les microorganismes cibles, car ils agissent sur les membranes cellulaires et ils devraient être plus vite dégradés sans produire de résidus.In spite of restricting rules, antibiotics are still used in aquaculture for prevention, especially during critical phases (early stages; metamorphosis; transfer), but also during on-growing. Besides improving husbandry techniques and prophylaxis, which is still possible, alternative methods are now available or in progress. Commercial probiotics for land animals are increasingly used for shrimp and fish farming, though every species or rearing condition should require particular experiments to determine the most suitable microbial preparations and doses. The variability in response to probiotics, and the lack of reliable data hinder the use of such practices in routine, indeed. These commercial preparations contain Lactobacillus, Bacillus, or other bacteria belonging to related genera, or yeast. They are not suitable for bivalve molluscs like scallops, oysters, and clams. Only few marine bacteria have been documented to protect efficiently larval bivalves against infection. However, many problems remain unsolved regarding legal authorization, production, preservation, and retailing of such preparations with gram-negative bacteria. Probiotics may display multiple effects, possibly combining bacterial antagonism to some effects on the host, e.g. stimulating immunity, or growth, though the modes of action are not fully understood. Probiotics are not as efficient as antibiotics in case of bacterial infection, however. Prebiotics like fructo-oligosaccharides (FOS) may also improve animal health, since they favor the establishment of commensal or beneficial gut microbiota. For instance, an experiment showed that FOS could increase growth of turbot fry. A third alternative may come from antimicrobial peptides, which have been described in many organisms. These peptides have wide spectra of action. They can kill gram negative and gram positive bacteria, envelop viruses, yeasts, and moulds. Several peptides were recently discovered in shrimp and oysters, where they are essential elements of innate defense, in the absence of acquired immunity. These molecules could advantageously replace antibiotics, since they are less susceptible to cause resistance in the target microorganisms, due to their direct action on membranes, and to their fast degradability, which avoid the accumulation of residues

Stylicins, a new family of antimicrobial peptides from the Pacific blue shrimp Litopenaeus stylirostris

The present study reports the characterization of Ls-Stylicin1, a novel antimicrobial peptide from the penaeid shrimp, Litopenoeus stylirostris. The predicted mature peptide of 82 residues is negatively charged (theoretical pl=5.0) and characterized by a proline-rich N-terminal region and a C-terminal region containing 13 cysteine residues. The recombinant Ls-Stylicin1 has been isolated in both monomeric and dimeric forms. Both display strong antifungal activity against Fusarium oxysporum (1.25 mu M < MIC <2.5 mu M), a pathogenic fungus of shrimp, but lower antimicrobial activity against Gram () bacteria, Vibrio sp. (40 mu M < MIC <80 mu M). However, rLs-Stylicin1 is able to agglutinate Vibrio pennaeicidae in vitro in agreement with its potent LPS-binding activity on immobilized LPS of V. penaeicidae (dissociation constant (K-d) of 9.6 x 10(-8) M). This molecule with no evident homology to other hitherto described antimicrobial peptides but identified herein several species of penaeid shrimp is thought to be the first member of a shrimp antimicrobial peptide family, which we termed stylicins. (C) 2009 Elsevier Ltd. All rights reserved

Monoclonal antibodies against Bonamia ostreae (Protozoa: Ascetospora), an intra-haemocytic parasite of flat oyster Ostrea edulis (Mollusca: Bivalvia)

The protozoan Bonamia ostreae (Ascetospora), a paasite of the flat oyster Ostrea edulis, was purified by differential and isopycnic centrifugations. Mice of the strain Balb/c were immunized with purified parasites and hybridomas were prepared by fusion of immunized mouse splenocytes with the mouse myeloma cell line P3-X63-Ag8-653. From the fusion, 12 clones were isolated and saved. The resulting antibodies were characterized with a solid phase radioimmunoassay (RIA). Antibodies from 7 clones reacted only with B. ostreae and not with normal host tissue. Although some of the antibiodies identified. One of these antibodies (20B2-1B12) has an apparent association constant of ca 3 X 10(8) M-1 and bound to a number of different sites on the parasite. These monoclonal antibodies should prove to be of great value as diagnostic and research tools