Differential immune gene response in gills, skin, and spleen of rainbow trout <i>Oncorhynchus mykiss</i> infected by<i> Ichthyophthirius multifiliis</i>

Infection of rainbow trout with the parasitic ciliate Ichthyopthirius multifiliis induces differential responses in gills, skin and spleen. A controlled experimental infection was performed and expression of immune-relevant genes in skin, gills, and spleen were recorded by qPCR at day 1 and 8 after parasite exposure. Infection induced a marked reaction involving regulation of innate and adaptive immune genes in rainbow trout at day 8 post-infection. The expression level of a total of 22 out of 24 investigated genes was significantly higher in gills compared to skin reflecting the more sensitive and delicate structure of gills. Especially pro-inflammatory cytokines IL-6, IL-17 C1, regulatory cytokines IL-4/13A, IL-10, TGFβ, complement factor C5, chemokines CK10, CK12, acute phase proteins (precerebellin, hepcidin) and immunoglobulins (IgM, IgT) displayed differential expression levels. The spleen, a central immune organ with no trace of the parasite, showed elevated expression of IgM, IgT, complement factor C5 and chemokine CK10 (compared to skin and gills directly exposed to the parasite), indicating an interaction between the infected surface sites and central immune organs. This communication could be mediated by chemokines CK10 and CK12 and cytokine IL-4/13A and may at least partly explain the establishment of a systemic response in rainbow trout against the parasite

Rainbow trout Oncorhynchus mykiss skin responses to salmon louse Lepeophtheirus salmonis: From copepodid to adult stage

The marine crustacean Lepeophtheirus salmonis (salmon louse) is a common ectoparasite of wild and farmed salmonids. The parasite has a complex ontogeny comprising eight instars. The planktonic copepodid stage settles on host skin and pass through five instars to reach the adult stage. The present study comprises an experimental infestation of Oncorhynchus mykiss (rainbow trout) with salmon lice and describes histopathology and host immune responses in skin beneath the louse at multiple time points encompassing all louse developmental stages. Each fish was exposed to 80 infective copepodids, a mean no. of 32 parasites reached the preadult I stage whereas a mean no. of 11 parasites reached the adult stage. A progression in the severity of cutaneous lesions was observed, and levels of immune gene transcripts at the attachment site revealed a dynamic response, initially related to innate immunity. Later, immune cells accumulated in the dermis concomitant with a moderate decrease in levels of transcripts characteristic of both innate and adaptive immune responses. The present study also demonstrates that the cutaneous immune response was mainly induced at lice affected sites, while non-affected skin resembled the skin of untreated control. This indicates that the skin cannot be regarded as a uniform organ and requires careful sampling at all salmon louse stages.publishedVersio

Insight from Molecular, Pathological, and Immunohistochemical Studies on Cellular and Humoral Mechanisms Responsible for Vaccine-Induced Protection of Rainbow Trout against <em>Yersinia ruckeri</em>

The immunological mechanisms associated with protection of vaccinated rainbow trout, Oncorhynchus mykiss, against enteric redmouth disease (ERM), caused by Yersinia ruckeri, were previously elucidated by the use of gene expression methodology and immunochemical methods. That approach pointed indirectly to both humoral and cellular elements being involved in protection. The present study correlates the level of protection in rainbow trout to cellular reactions in spleen and head kidney and visualizes the processes by applying histopathological, immunohistochemical, and in situ hybridization techniques. It was shown that these cellular reactions, which were more prominent in spleen than in head kidney, were associated with the expression of immune-related genes, suggesting a Th2-like response. Y. ruckeri, as shown by in situ hybridization (ISH), was eliminated within a few days in vaccinated fish, whereas nonprotected fish still harbored bacteria for a week after infection. Vaccinated fish reestablished normal organ structure within a few days, whereas nonprotected fish showed abnormalities up to 1 month postinfection. Protection in the early phase of infection was mainly associated with the expression of genes encoding innate factors (complement factors, lysozyme, and acute phase proteins), but in the later phase of infection, increased expression of adaptive immune genes dominated. The histological approach used has shown that the cellular changes correlated with protection of vaccinated fish. They comprised transformation of resident cells into macrophage-like cells and increased occurrence of CD8α and IgM cells, suggesting these cells as main players in protection. Future studies should investigate the causality between these factors and protection

Genetic responses of the marine copepod Acartia tonsa (Dana) to heat shock and epibiont infestation

Expression of stress-related genes was investigated in the marine copepod Acartia tonsa in relation to heat shock at two different salinities (10 and 32‰), and it was furthermore investigated whether experimentally induced epibiont infestation led to elevated expression of stress-related genes. Expression of the genes ferritin, Hsp90 and Hsp70 were analyzed in adult copepods by conducting reverse transcription-quantitative real time PCR (RT-qPCR). The expression of Hsp70 and Hsp90 was significantly up-regulated after heat shock and the expression levels were higher in copepods cultivated at 10‰ salinity seawater than in copepods cultivated at 32‰. Significant up-regulation of ferritin (3.3 fold increase) was observed as a response to infestation with the epibiotic euglenid Colacium vesiculosum. Results suggest that (i) A. tonsa responds more pronounced to thermal shock when cultivated in low salinity seawater (10‰) as compared to optimal salinity seawater (32‰) and (ii) epibiont infestation does cause a measureable physiological response in the host

Cannabidiol effects on behaviour and immune gene expression in zebrafish (<i>Danio rerio</i>)

Various preparations and extracts of the plant Cannabis sativa (family cannabaceae) are used as herbal medicinal drugs against a series of disorders but the plant contains a wide series of pharmacologically active components which may confound evaluation of drug effects. In order to differentiate specific effects of the individual constituents on specific functions in the organism we advocate for controlled studies on specified constituents and their impact on the vertebrate organism. One of the dominating Cannabis constituents, delta(9)-tetrahydrocannabinol (THC), has previously been studied in depth whereas information on another main ingredient cannabidiol (CBD) is limited. We have performed a controlled study on CBD and its effect using an experimental zebrafish model. CDB treatment of zebrafish for 30 min affected mobility of the fish by decreasing swimming speed and swimming distance. In addition, out of 23 immune related genes studied it was shown that expression of two genes il1b and il17a/f2 were up-regulated and four genes, tgfba, ighm, cd4-1, and s100a10b were significantly down-regulated following CBD treatment. The study indicated that CBD affects motility and immunity of the vertebrate host