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The DAFINET workshop on Sustainable fish health control was organized in collaboration with Parafishcontrol and BANGFISH on October 23, 2018. In connection to the workshop the University of Copenhagen hosted a five day workshop on fish diseases. The 40 workshop participants comprised PFC partners and colleagues from various countries (Norway, Bangladesh, India, China, Indonesia, Italy, Denmark). The scientific fields touched upon ranged from PFC parasites (salmon lice, white spot disease parasite Ichthyophthirius multifiliis, zoonotic nematode Contracaecum osculatum), and environmental issues (arsenic substances and antibiotics in fish ponds), pigments in fish flesh, bacterial and viral infections of rainbow trout. The host species discussed ranged from Atlantic salmon and rainbow trout to tilapia and Asian catfish. Sonal Patel from Norway and Morten Limborg from Denmark presented new results and ideas from novel types of approaches in fish disease research – the holistic view on both host, microbiome and the environment. The connected fish disease course over five days focused on both basic classical techniques and more advanced methodologies such as ELISA, immunohistochemistry, histology, PCR, qPCR and DNA sequencing. The course addressed parasitic, bacterial and viral diseases in wild and farmed fish. The photo shows seven course participants from Bangladesh with professor Kurt Buchmann (KU) in the lab.

Hints on T cell responses in a fish-parasite model: Enteromyxum leei induces differential expression of T cell signature molecules depending on the organ and the infection status. Parasites & Vectors 11:443. doi.org/10.1186/s13071-018-3007-1

Enteromyxum leei is a myxozoan parasite that produces a slow-progressing intestinal disease. This parasite invades the paracellular space of the intestinal epithelium and progresses from the posterior to the anterior intestine. The aim of the present study was to gain insights into fish T cell responses in the gilthead sea bream-E. leei infection model using a PCR-array with 30 signature molecules for different leukocyte responses in head kidney, spleen, anterior and posterior intestine.
The PCR-array results suggest that E. leei induced migration of T cells from head kidney to intestines where T_H1, CTL and T_H17 profiles were activated and kept in balance by the upregulation of regulatory cytokines. These results were partially validated by the use of cross-reacting antibodies and BrdU immunostaining to monitor proliferation. Zap70 immunostaining supported the increased number of T cells in the anterior intestine detected by gene expression, but double staining with BrdU did not show active proliferation of this cell type at a local level, supporting the migration from lymphohaematopoietic tissues to the site of infection. Global analyses of the expression profiles revealed a clear separation between infected and exposed, but non-infected fish, more evident in the target organ. Exposed, non-infected animals showed an intermediate phenotype closer to the control fish.
These results evidence a clear modulation of the T cell response of gilthead sea bream upon E. leei infection. The effects occurred both at local and systemic levels, but the response was stronger and more specific at the site of infection, the intestine. Altogether, this research poses a promising basis to understand the response against this important parasite and establish effective preventive or palliative measures.

Check out all ParaFishControl publications here

Dysregulation of B Cell Activity During Proliferative Kidney Disease in Rainbow Trout. Frontiers in Immunology. doi.org/10.3389/fimmu.2018.01203

Proliferative kidney disease (PKD) is a widespread disease caused by the endoparasite Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea). Clinical disease, provoked by the proliferation of extrasporogonic parasite stages, is characterized by a chronic kidney pathology with underlying transcriptional changes indicative of altered B cell responses and dysregulated T-helper cell-like activities. Despite the relevance of PKD to European and North American salmonid aquaculture, no studies, to date, have focused on further characterizing the B cell response during the course of this disease. Thus, in this work, we have studied the behavior of diverse B cell populations in rainbow trout (Oncorhynchus mykiss) naturally infected with T. bryosalmonae at different stages of preclinical and clinical disease. Our results show a clear upregulation of all trout immunoglobulins (Igs) (IgM, IgD, and IgT) demonstrated by immunohistochemistry and Western blot analysis, suggesting the alteration of diverse B cell populations that coexist in the infected kidney. Substantial changes in IgM, IgD, and IgT repertoires were also identified throughout the course of the disease further pointing to the involvement of the three Igs in PKD through what appear to be independently regulated mechanisms. Thus, our results provide strong evidence of the involvement of IgD in the humoral response to a specific pathogen for the first time in teleosts. Nevertheless, it was IgT, a fish-specific Ig isotype thought to be specialized in mucosal immunity, which seemed to play a prevailing role in the kidney response to T. bryosalmonae. We found that IgT was the main Ig coating extrasporogonic parasite stages, IgT+ B cells were the main B cell subset that proliferated in the kidney with increasing kidney pathology, and IgT was the Ig for which more significant changes in repertoire were detected. Hence, although our results demonstrate a profound dysregulation of different B cell subsets during PKD, they point to a major involvement of IgT in the immune response to the parasite. These results provide further insights into the pathology of PKD that may facilitate the future development of control strategies.

Check out all ParaFishControl publications here