Desempenho e hematologia de tilápias-do-nilo alimentadas com Saccharomyces cerevisiae e vacinadas contra Streptococcus agalactiae

O objetivo deste trabalho foi avaliar a inter-relação entre a suplementação alimentar com parede celular de Saccharomyces cerevisae e a vacinação contra Streptococcus agalactiae e seu efeito sobre o desempenho produtivo e as variáveis hematológicas de tilápia-do-nilo (Oreochromis niloticus). Oitenta e quatro tilápias-do-nilo foram distribuídas em 12 caixas de fibra (n=7), em arranjo fatorial 2x2x3, correspondente a dois níveis de suplementação com parede celular de levedura, dois tipos de inoculação e três tempos de avaliação. Os peixes foram alimentados durante 77 dias. A vacinação dos peixes foi realizada 60 dias após o início da alimentação. Quinze dias após a vacinação, todos os peixes foram submetidos ao desafio com cepa viva de S. agalactiae, e 6, 24 e 48 horas após o desafio, o sangue foi colhido da veia caudal para avaliações. Peixes alimentados com ração suplementada apresentam maior ganho de peso e taxa de crescimento específico, e a interação entre os efeitos da dieta e da vacinação resulta em maiores taxas de hematócrito, hemoglobina e leucócitos

Ontogeny of humoral immune parameters in fish

The first appearance of IgM in lymphocytes varies considerably among fish species. Generally, the first appearance of B-lymphocytes and immunoglobulins is late in marine species compared to fresh water species, and larvae have reached about 20–30 mm in length when IgM is first expressed. Rainbow trout and channel catfish show the first appearances of surface IgM at about 1 week after hatching. Marine species like the sea bass, spotted wolffish and cod show IgM positive lymphocytes 1–10 weeks after hatching. Transfer of maternal antibody to eggs and embryos has been demonstrated in several species. Examples are plaice, tilapia, carp, sea bass and salmon, but not cod. The ontogeny of complement component C3 has been studied in Atlantic halibut (Hippoglossus hippoglossus L.), Atlantic cod (Gadus morhua L.) and the spotted wolffish (Anarhichas minor O.). By Western blotting experiments C3 was found in unfertilised eggs in the spotted wolffish indicating a maternal transfer. RT-PCR analysis revealed C3 mRNA transcripts from 290 d° in spotted wolffish eggs. Using immunohistochemistry and in situ hybridisation, C3 was found in liver, brain, kidney and muscle of cod larvae 2 days post-hatching and in intestines, pancreas, heart and gills at different stages of larval development. Also, C3 was detectable in halibut larvae in yolk sac, muscle, liver, brain, chondrocytes, spinal chord, eye, heart, intestines and kidney. These studies suggest that complement may play a role in generation of different organs, not only in the defence against invading pathogens. Lysozyme is a bactericidal enzyme present in mucus, lymphoid tissue and serum of most fish species, but not in cod and wolffish. The enzyme has been detected in oocytes, fertilised eggs and larval stages of fish species like coho salmon, sea bass and tilapia. The activity of other enzymes like the cathepsins has been described in eggs and larvae of sea bass, cod and salmonids. Cathepsins may have a bactericidal role in the skin of fish. Lectins are carbohydrate-binding proteins that interact with pathogenic surface structures that result in opsonization, phagocytosis or activation of complement. Lectins have been isolated from the eggs of various fish species

Immunostimulation of larvae and juveniles of cod, Gadus morhua L.

Cod larval culture is currently hampered by high mortalities in the first 2–3 weeks after hatching, often due to infectious diseases. The immune system of cod is not fully competent until 2–3 months after hatching. Conventional vaccination is, therefore, not of value before this time, and the larvae are wholly reliant on non-specific parameters for their defence against infection. A range of substances, generally derived from bacterial, fungal or plant origin, can activate these non-specific parameters. During three hatching seasons, 2001–2003, at the Marine Institute's Experimental Station, Stadur, Grindavik, Iceland, the effects of several immunostimulants on survival and disease resistance of cod larvae and juveniles were examined. Both bathing treatments and administration in the feed were used. One of these substances, lipopolysaccharide (LPS), isolated from the bacterium Aeromonas salmonicida (ssp. salmonicida or achromogenes), appeared in some instances to improve survival and have a beneficial effect on disease resistance. Other substances tested had limited effects. The results emphasize the need for further work in this field

Establishment of killer whale (Orcinus orca) primary fibroblast cell cultures and their transcriptomic responses to pollutant exposure

Populations of killer whale (Orcinus orca) contain some of the most polluted animals on Earth. Yet, the knowledge on effects of chemical pollutants is limited in this species. Cell cultures and in vitro exposure experiments are pertinent tools to study effects of pollutants in free-ranging marine mammals. To investigate transcriptional responses to pollutants in killer whale cells, we collected skin biopsies of killer whales from the Northern Norwegian fjords and successfully established primary fibroblast cell cultures from the dermis of 4 out of 5 of them. Cells from the individual with the highest cell yield were exposed to three different concentrations of a mixture of persistent organic pollutants (POPs) that reflects the composition of the 10 most abundant POPs found in Norwegian killer whales (p,p’-DDE, trans-nonachlor, PCB52, 99, 101, 118, 138, 153, 180, 187). Transcriptional responses of 13 selected target genes were studied using digital droplet PCR, and whole transcriptome responses were investigated utilizing RNA sequencing. Among the target genes analysed, CYP1A1 was significantly downregulated in the cells exposed to medium (11.6 µM) and high (116 µM) concentrations of the pollutant mixture, while seven genes involved in endocrine functions showed a non-significant tendency to be upregulated at the highest exposure concentration. Bioinformatic analyses of RNA-seq data indicated that 13 and 43 genes were differentially expressed in the cells exposed to low and high concentrations of the mixture, respectively, in comparison to solvent control. Subsequent pathway and functional analyses of the differentially expressed genes indicated that the enriched pathways were mainly related to lipid metabolism, myogenesis and glucocorticoid receptor regulation. The current study results support previous correlative studies and provide cause-effect relationships, which is highly relevant for chemical and environmental management