Publications by authors named "Per W Kania"

34 Publications

(Apicomplexa) in in Europe with a potential for spread.

Int J Parasitol Parasites Wildl 2021 Aug 2;15:270-275. Epub 2021 Jul 2.

Laboratory of Aquatic Pathobiology, Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.

Four specimens of mallard () shot by local hunters (December 2020 to January 2021 along the eastern coastline of the island of Bornholm in the Baltic Sea) were diagnosed with a heavy load of sarcocysts in the musculature. Morphometric and molecular diagnosis based on rDNA (18 S, ITS1, 28 S) of parasites recovered from two of the birds revealed the causative pathogen to be . We further present novel sequences for the entire 5.8 S and ITS2 for this species. Elongate cysts (mean length 5.25 (SD 0.6) mm, width 1.37 (SD 0.2) mm) were recorded in all parts of the striated skeletal musculature of the birds. The main part (72%) of the 2585 cysts in one female mallard was located in the outer superficial pectoral musculature, with 11% in the inner pectoral musculature. Minor but significant parts were found in the dorsal, ventral abdominal, neck and head, legs, hand and arm (wing) musculature. No cysts were found in the smooth musculature. Each cyst contained a median of 3.2 mio bradyzoites indicating that more than 8 billion bradyzoites are available for infection of one or more predators/scavengers ingesting the bird. Bradyzoites (median length 13.5 μm (range 12.1-14.5) and median width 2.66 μm (range 2.1-3.3)) were highly resistant to proteinase treatment, which secures the passage through the stomach of the predator to its intestine where wall penetration takes place. One of the birds was ringed (tagged) in Sweden Island Øland in the Baltic Sea two years before being shot. This is documenting immigration of mallards from northern locations. The parasite species was originally described in North America in 1893 and was commonly reported in this region during the 20th century but not in Europe. Recent cases from Norway, Finland, Lithuania, Poland, UK and Hungary suggest that the species may be spreading geographically. Experienced duck hunters with a 40 years record of hunting on the island reported that this type of infection unprecedented. The final host is reported to be canines (fox, raccoon dog), skunk and mustelids, including ermines and American mink. Presence of these hosts in Europe may allow establishment of the life cycle and further colonization of the local duck populations which calls for implementation of a survey program in Europe.
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http://dx.doi.org/10.1016/j.ijppaw.2021.06.007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8264532PMC
August 2021

A Major QTL for Resistance to in Rainbow Trout.

Front Genet 2020 29;11:607558. Epub 2020 Dec 29.

Laboratory of Aquatic Pathobiology, Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Genetic selection of disease resistant fish is a major strategy to improve health, welfare and sustainability in aquaculture. Mapping of single nucleotide polymorphisms (SNP) in the fish genome may be a fruitful tool to define relevant quantitative trait loci (QTL) and we here show its use for characterization of resistant rainbow trout (). Fingerlings were exposed to the pathogen serotype O1 in a solution of 1.5 × 10 cfu/ml and observed for 14 days. Disease signs appeared 3 days post exposure (dpe) whereafter mortality progressed exponentially until 6 dpe reaching a total mortality of 55% within 11 days. DNA was sampled from all fish - including survivors - and analyzed on a 57 k Affymetrix SNP platform whereby it was shown that disease resistance was associated with a major QTL on chromosome 21 (Omy 21). Gene expression analyses showed that diseased fish activated genes associated with innate and adaptive immune responses. The possible genes associated with resistance are discussed.
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http://dx.doi.org/10.3389/fgene.2020.607558DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7802751PMC
December 2020

Immune gene expression and genome-wide association analysis in rainbow trout with different resistance to Yersinia ruckeri infection.

Fish Shellfish Immunol 2020 Nov 11;106:441-450. Epub 2020 Aug 11.

Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C., Denmark.

Selective breeding programmes involving marker assisted selection of innately pathogen resistant strains of rainbow trout rely on reliable controlled infection studies, extensive DNA typing of individual fish and recording of expression of relevant genes. We exposed juvenile rainbow trout (6 h bath to 2.6 × 10 CFU mL) to the fish pathogen Yersinia ruckeri serotype O1, biotype 2, eliciting Enteric Red Mouth Disease ERM, and followed the disease progression over 21 days. Cumulative mortality reached 42% at 12 days post challenge (dpc) after which no disease signs were recorded. All fish were sampled for DNA-typing (50 k SNP chip, Affymetrix®) throughout the course of infection when they showed clinical signs of disease (susceptible fish) or at day 21 when fish showed no clinical signs of disease (survivors - resistant fish). Genome-wide association analyses of 1027 trout applying single nucleotide polymorphisms (SNPs) as markers revealed an association between traits (susceptible/resistant) and certain regions of the trout genome. It was indicated that multiple genes are involved in rainbow trout resistance towards ERM whereby it is considered a polygenic trait. A corresponding trout group was kept as non-exposed controls and a comparative expression analysis of central innate and adaptive immune genes in gills, spleen and liver was performed for three fish groups: 1) moribund trout exhibiting clinical signs 7 dpc (CS), 2) exposed fish without clinical signs at the same sampling point (NCS) and 3) surviving fish at 21 dpc (survivors). Immune genes encoding inflammatory cytokines (IL-1β, IL-2A, IL-6A, IL-8, IL-10A, IL-12, IL-17A/F2A, IL-17C1, IL-17C2, IL-22, IFNγ, TNFα), acute phase reactants (SAA, C3, cathelicidins, lysozyme) were expressed differently in CS and NCS fish. Correlation (negative or positive) between expression of genes and bacterial load suggested involvement of immune genes in protection. Down-regulation of adaptive immune genes including IgDm, IgDs, IgT and TCR-β was seen primarily in CS and NCS fish whereas survivors showed up-regulation of effector molecule genes such as cathelicidins, complement and lysozyme suggesting their role in clearing the infection. In conclusion, SNP analyses indicated that ERM resistance in rainbow trout is a multi-locus trait. The gene expression in surviving fish suggested that several immune genes are associated with the trait conferring resistance.
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http://dx.doi.org/10.1016/j.fsi.2020.07.023DOI Listing
November 2020

Anisakid nematode larvae in the liver of Atlantic cod Gadus morhua L. from West Greenland.

Parasitol Res 2020 Oct 13;119(10):3233-3241. Epub 2020 Jul 13.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 7, DK-1870, Frederiksberg C, Denmark.

Anisakid nematode larvae occur frequently in the liver of Atlantic cod, but merely few infection data from cod in waters around Greenland exist. The present study reports the occurrence of third-stage anisakid larvae in the livers of 200 Atlantic cod caught on fishing grounds along the West coast of Greenland (fjord systems of Maniitsoq) in May, June, August and September 2017. Classical and molecular helminthological techniques were used to identify the nematodes. A total of 200 cod livers were examined, and 194 were infected with third-stage nematode larvae (overall prevalence of infection 97%) with a mean intensity of 10.3 (range between 1 and 44 parasites per fish). Prevalences recorded were 96% for Anisakis simplex (s.l.), 55% for Pseudoterranova decipiens (s.l.) and 8% for Contracaecum osculatum (s.l.). Sequencing the mtDNA cox2 from 8 out of 23 these latter larvae conferred these to C. osculatum sp. B. A clear seasonal variation was observed, with a rise in A. simplex (s.l.) and P. decipiens (s.l.) occurrence in June and August and a decline in September. The study may serve as a baseline for future investigations using the three anisakids as biological indicators in Greenland waters.
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http://dx.doi.org/10.1007/s00436-020-06807-zDOI Listing
October 2020

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

Fish Shellfish Immunol 2020 Aug 15;103:200-210. Epub 2020 May 15.

SLRC - Sea Lice Research Centre, Department of Biological Sciences, University of Bergen, Thormøhlensgt. 55, Pb. 7803, 5020, Bergen, Norway. Electronic address:

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.
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http://dx.doi.org/10.1016/j.fsi.2020.05.014DOI Listing
August 2020

Endoparasitic helminths in Baltic salmon Salmo salar: ecological implications.

Dis Aquat Organ 2019 Sep;135(3):193-199

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C 1870, Denmark.

Parasites in fish are ecological indicators, as they reflect the host's migration routes, feeding behavior and immune status. We performed a parasitological investigation of sea-running Baltic salmon to study the use of parasites as indicators for this fish stock. The host-a strain of Atlantic salmon Salmo salar-has been isolated for several millennia in the semi-enclosed brackish Baltic Sea, with limited migration to and from the North Sea. Twenty-four salmon (total body weight: 4.2-14.2 kg; total body length: 80-105 cm) were caught by spoon bait in the southern Baltic Sea during feeding migrations, necropsied shortly afterwards and internal organs subjected to parasitological investigation focusing on endoparasitic helminths. The pyloric region was heavily parasitized by the cestode Eubothrium crassum (100% prevalence; intensity: 97-273 parasites per infected fish), reflecting a diet of smaller pelagic fishes. The stomach contained the hemiurid digeneans Brachyphallus crenatus (95.8% prevalence; intensity: 8-151) and Hemiurus luehei (58.3% prevalence; intensity: 2-13), indicating a diet of clupeids. Schistocephalus solidus (25% prevalence; intensity: 1-2), liberated from ingested sticklebacks, the acanthocephalan Echinorhynchus truttae (54% prevalence; intensity: 1-13) and the adult nematode Hysterothylacium aduncum (29% prevalence; intensity: 1-13) were found in the intestine. The liver was parasitized by third-stage nematode larvae of Contracaecum osculatum (45.8% prevalence; intensity: 1-4), but these were growth-stunted and encapsulated. The parasite fauna differs markedly from salmon in North Atlantic waters, and the lack of purely marine parasite species indicates that the Baltic salmon has remained in the Baltic Sea during its life history.
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http://dx.doi.org/10.3354/dao03391DOI Listing
September 2019

Transcriptomic analysis of Baltic cod (Gadus morhua) liver infected with Contracaecum osculatum third stage larvae indicates parasitic effects on growth and immune response.

Fish Shellfish Immunol 2019 Oct 13;93:965-976. Epub 2019 Aug 13.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

High infection levels due to third-stage larvae of the anisakid nematode Contracaecum osculatum have been documented in cod from the eastern part of the Baltic sea during the latest decades. The nematode larvae mainly infect the liver of Baltic cod and prevalence of infection has reached 100% with a mean intensity up to 80 parasites per host in certain areas and size classes. Low condition factors of the cod have been observed concomitant with the rise in parasite abundance suggesting a parasitic effect on growth parameters. To investigate any association between parasite infection and physiological status of the host we performed a comparative transcriptomic analysis of liver obtained from C. osculatum infected and non-infected cod. A total of 47,025 predicted gene models showed expression in cod liver and sequences corresponding to 2084 (4.43%) unigenes were differentially expressed in infected liver when compared to non-infected liver. Of the differentially expressed unigenes (DEGs) 1240 unigenes were up-regulated while 844 unigenes were down-regulated. The Gene Ontology (GO) enrichment analysis showed that 1304 DEGs were represented in cellular process and single-organism process, cell and cell part, binding and catalytic activity. As determined by the Kyoto Encyclopedia of Gene and Genomes (KEGG) Pathways analysis, 454 DEGs were involved in 138 pathways. Ninety-seven genes were related to metabolic pathways including carbohydrate, lipid, and amino acid metabolism. Thirteen regulated genes were playing a role in immune response such as Toll-like receptor signaling, NOD-like receptor signaling, RIG-I-like receptor signalling and thirty-six genes were associated with growth processes. This indicates that the nematode infection in Baltic cod may affect on molecular mechanisms involving metabolism, immune function and growth.
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http://dx.doi.org/10.1016/j.fsi.2019.08.034DOI Listing
October 2019

Differential immune gene response in gills, skin, and spleen of rainbow trout Oncorhynchus mykiss infected by Ichthyophthirius multifiliis.

PLoS One 2019 20;14(6):e0218630. Epub 2019 Jun 20.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

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.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0218630PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6586319PMC
February 2020

A pentavalent vaccine for rainbow trout in Danish aquaculture.

Fish Shellfish Immunol 2019 May 6;88:344-351. Epub 2019 Mar 6.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark.

Mariculture in Denmark is based on production of rainbow trout grown two years in fresh water followed by one growth season in sea cages. Although the majority of rainbow trout are vaccinated against the most serious bacterial pathogens - Aeromonas salmonicida subsp. salmonicida, Vibrio anguillarum and Yersinia ruckeri, by the use of commercially available vaccines, disease outbreaks requiring treatment with antibiotics still occur. The present study tested the potential of a new experimental multicomponent vaccine that is based on local bacterial strains, isolated from rainbow trout in Danish waters, and thus custom-designed for Danish rainbow trout mariculture. The vaccination with the multicomponent vaccine resulted in protection against three relevant bacterial diseases (yersiniosis, furunculosis, vibriosis) under experimental conditions. We showed that i.p. injection of the vaccine induced specific antibody responses in trout against the different bacterial antigens and regulated expression of genes encoding SAA, C3, IL-1β, IL-6, IL-8, IgD and MHCII.
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http://dx.doi.org/10.1016/j.fsi.2019.03.001DOI Listing
May 2019

CK11, a Teleost Chemokine with a Potent Antimicrobial Activity.

J Immunol 2019 02 4;202(3):857-870. Epub 2019 Jan 4.

Animal Health Research Centre, National Institute for Agricultural and Food Research, Valdeolmos 28130, Madrid, Spain;

CK11 is a rainbow trout () CC chemokine phylogenetically related to both mammalian CCL27 and CCL28 chemokines, strongly transcribed in skin and gills in homeostasis, for which an immune role had not been reported to date. In the current study, we have demonstrated that CK11 is not chemotactic for unstimulated leukocyte populations from central immune organs or mucosal tissues but instead exerts a potent antimicrobial activity against a wide range of rainbow trout pathogens. Our results show that CK11 strongly inhibits the growth of different rainbow trout Gram-positive and Gram-negative bacteria, namely , subsp. , and and a parasitic ciliate Similarly to mammalian chemokines and antimicrobial peptides, CK11 exerted its antimicrobial activity, rapidly inducing membrane permeability in the target pathogens. Further transcriptional studies confirmed the regulation of CK11 transcription in response to exposure to some of these pathogens in specific conditions. Altogether, our studies related to phylogenetic relations, tissue distribution, and biological activity point to CK11 as a potential common ancestor of mammalian CCL27 and CCL28. To our knowledge, this study constitutes the first report of a fish chemokine with antimicrobial activity, thus establishing a novel role for teleost chemokines in antimicrobial immunity that supports an evolutionary relationship between chemokines and antimicrobial peptides.
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http://dx.doi.org/10.4049/jimmunol.1800568DOI Listing
February 2019

Transcriptomic analysis of immunity in rainbow trout (Oncorhynchus mykiss) gills infected by Ichthyophthirius multifiliis.

Fish Shellfish Immunol 2019 Mar 1;86:486-496. Epub 2018 Dec 1.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

The parasite Ichthyophthirius multifiliis infecting skin, fins and gills of a wide range of freshwater fish species, including rainbow trout, is known to induce a protective immune response in the host. Although a number of studies have reported activation of several immune genes in infected fish host, the immune response picture is still considered incomplete. In order to address this issue, a comparative transcriptomic analysis was performed on infected versus uninfected rainbow trout gills and it showed that a total of 3352 (7.2%) out of 46,585 identified gene sequences were significantly regulated after parasite infection. Of differentially expressed gene sequences, 1796 genes were up-regulated and 1556 genes were down-regulated. These were classified into 61 Gene Ontology (GO) terms and mapped to 282 reference canonical pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Infection of I. multifiliis induced a clear differential expression of immune genes, related to both innate and adaptive immunity. A total of 268 (6.86%) regulated gene sequences were known to take part in 16 immune-related pathways. These involved pathways related to the innate immunity such as the Chemokine signaling pathway, Platelet activation, Toll-like receptor signaling pathway, NOD-like receptor signaling pathway, and Leukocyte transendothelial migration. Elevated transcription of genes encoding the TLR 8 gene and chemokines (CCL4, CCL19, CCL28, CXCL8, CXCL11, CXCL13, CXCL14) was recorded indicating their roles in recognition of I. multifiliis and subsequent induction of the inflammatory response, respectively. A number of upregulated genes in infected gills were associated with antigen processing/presentation and T and B cell receptor signaling (including B cell marker CD22 involved in B cell development). Overall the analysis supports the notion that I. multifiliis induces a massive and varied innate response upon which a range of adaptive immune responses are established which may contribute to the long lasting protection of immunized rainbow trout.
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http://dx.doi.org/10.1016/j.fsi.2018.11.075DOI Listing
March 2019

Cannabidiol effects on behaviour and immune gene expression in zebrafish (Danio rerio).

PLoS One 2018 31;13(7):e0200016. Epub 2018 Jul 31.

Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C., Denmark.

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.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0200016PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6067702PMC
January 2019

Effect of oral booster vaccination of rainbow trout against Yersinia ruckeri depends on type of primary immunization.

Fish Shellfish Immunol 2019 Feb 31;85:61-65. Epub 2017 Oct 31.

Department of Veterinary and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

Vaccination of rainbow trout against Enteric Redmouth Disease (ERM) caused by Yersinia ruckeri can be successfully performed by administering vaccine (a bacterin consisting of formalin killed bacteria) by immersion, bath or injection. Booster immunization is known to increase the protection of fish already primed by one of these vaccination methods. Oral vaccination of trout (administering vaccine in feed) is an even more convenient way of presenting antigen to the fish but the effect of an oral booster has not previously been described in detail. The present work describes to what extent protection may be enhanced by oral boostering following priming with different administration methods. The study confirms that vaccination by 30 s dip into a bacterin (diluted 1:10) may confer a significant protection compared to non-vaccinated fish. The immunity may be optimized by booster immunization either provided as dip (most effective), bath (less effective) or orally (least effective). Oral immunization may be used as booster after dip but applied as a single oral application it induced merely a slight and statistically non-significant response. It is noteworthy that primary oral immunization followed by an oral booster vaccination showed a trend for an even weaker response. It should be investigated if continued exposure to a low antigen concentration - as performed by two oral immunizations - may induce tolerance to the pathogen and thereby leave the fish more vulnerable.
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http://dx.doi.org/10.1016/j.fsi.2017.10.049DOI Listing
February 2019

Molecular diversity of avian schistosomes in Danish freshwater snails.

Parasitol Res 2016 Mar 17;115(3):1027-37. Epub 2015 Nov 17.

Section for Parasitology and Aquatic Diseases, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Dyrlægevej 100, DK-1870, Frederiksberg C, Copenhagen, Denmark.

Avian schistosomes are widespread parasites of snails and waterfowl and may cause cercarial dermatitis (swimmer's itch) in humans, a disease that is frequently reported in European countries. These parasites are known to occur in Denmark, but here, we applied a new approach using molecular tools to identify the parasites at species level. In order to do that, 499 pulmonate freshwater snails (Radix sp., Lymnaea stagnalis, Stagnicola sp. and Planorbarius corneus) were sampled from 12 lakes, ponds, and marshes in the greater Copenhagen area. Avian schistosome cercariae were identified by microscopy and subjected to molecular investigation by sequencing and phylogenetic analysis of the 5.8S and ITS2 ribosomal DNA for species identification. Additionally, snail hosts belonging to the genus Radix were identified by sequencing and phylogenetic analysis of partial ITS2 ribosomal DNA. Three out of 499 snails shed different species of Trichobilharzia cercariae: Trichobilharzia szidati was isolated from L. stagnalis, Trichobilharzia franki from Radix auricularia and Trichobilharzia regenti from Radix peregra. In the light of the public health risk represented by bird schistosomes, these findings are of concern and, particularly, the presence of the potentially neuro-pathogenic species, T. regenti, in Danish freshwaters calls for attention.
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http://dx.doi.org/10.1007/s00436-015-4830-3DOI Listing
March 2016

Effects of adjuvant Montanide™ ISA 763 A VG in rainbow trout injection vaccinated against Yersinia ruckeri.

Fish Shellfish Immunol 2015 Dec 19;47(2):797-806. Epub 2015 Oct 19.

Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

Enteric redmouth disease (ERM) caused by the fish pathogen Yersinia ruckeri is a major threat to freshwater production of rainbow trout (Oncorhynchus mykiss) throughout all life stages. Injection vaccination of rainbow trout against Y. ruckeri infection has been shown to confer better protection compared to the traditionally applied immersion vaccination. It may be hypothesized, based on experience from other vaccines, that adjuvants may increase the protective level of ERM injection vaccines even more. Controlled comparative vaccination studies have been performed to investigate effects of the oil adjuvant Montanide™ ISA 763 A VG (Seppic) when added to an experimental Y. ruckeri bacterin (containing both biotype 1 and 2 of serotype O1). A total of 1000 fish with mean weight 19 g was divided into five different groups (in duplicated tanks 2 × 100 fish per group) 1) non-vaccinated control fish (NonVac), 2) fish injected with a commercial vaccine (AquaVac(®) Relera™) (ComVac), 3) fish injected with an experimental vaccine (ExpVac), 4) fish injected with an experimental vaccine + adjuvant (ExpVacAdj) and 5) fish injected with adjuvant alone (Adj). Injection of the experimental vaccine (both adjuvanted and non-adjuvanted) induced a significantly higher antibody (IgM) level, increased occurrence of IgM(+) cells in spleen tissue and significant up-regulation of several immune genes. Additional experiments using a higher challenge dosage suggested an immune enhancing effect of the adjuvant as the challenge produced 100% mortality in the NonVac group, 60% mortality in both of ComVac and Adj groups and only 13 and 2.5% mortalities in the ExpVac and the ExpVacAdj groups, respectively.
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http://dx.doi.org/10.1016/j.fsi.2015.10.023DOI Listing
December 2015

Eye fluke infection status in Baltic cod, Gadus morhua, after three decades and their use as ecological indicators.

Acta Parasitol 2015 Sep;60(3):423-9

Eye flukes of the genus Diplostomum were recorded with a prevalence of 7.4% and a mean intensity of 11.9 (range 1-75) parasites per fish in eye lenses of a total of 188 Atlantic cod, Gadus morhua, sampled in the Southeastern Baltic Sea from March 2013 to February 2014. A slight decrease of infection level, among the fish with body length ranging from 30 to 89.5 cm, was found when data were compared to a corresponding survey in the 1980s. Due to imprecise species identification of eye flukes based on morphometric analyses we present, as a baseline for further studies, rDNA sequences from a subsample of 19 eye flukes based on sequencing of a part of 18S, ITS-1, 5.8S, ITS-2 and part of 28S. We discuss the use of eye fluke recordings in Baltic cod as an environmental indicator due to the dependence of the parasite's life cycle on biotic (occurrence of snail and bird hosts) and abiotic (temperature and salinity) parameters.
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http://dx.doi.org/10.1515/ap-2015-0058DOI Listing
September 2015

Comparative evaluation of infection methods and environmental factors on challenge success: Aeromonas salmonicida infection in vaccinated rainbow trout.

Fish Shellfish Immunol 2015 Jun 14;44(2):485-95. Epub 2015 Mar 14.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark. Electronic address:

When testing vaccine-induced protection an effective and reliable challenge method is a basic requirement and we here present a comparative study on different challenge methods used for infection of rainbow trout Oncorhynchus mykiss with Aeromonas salmonicida, a bacterial pathogen eliciting furunculosis. Fish were vaccinated with three different adjuvanted trivalent vaccines containing formalin killed A. salmonicida, Vibrio anguillarum O1 and O2a. These were 1) the commercial vaccine Alpha Ject 3000, 2) an experimental vaccine with water in paraffin oil adjuvant, 3) an experimental vaccine with water in paraffin oil in water adjuvant. Fish were then exposed to A. salmonicida challenge using i.p. injection, cohabitation in freshwater, cohabitation in saltwater (15 ppt) or combined fresh/saltwater cohabitation. Cohabitation reflects a more natural infection mode and was shown to give better differentiation of vaccine types compared to i.p. injection of live bacteria. The latter infection mode is less successful probably due to the intra-abdominal inflammatory reactions (characterized in this study according to the Speilberg scale) induced by i.p. vaccination whereby injected live bacteria more effectively become inactivated at the site of injection. Compared to cohabitation in freshwater, cohabitation in saltwater was less efficient probably due to reduced survivability of A. salmonicida in saltwater, which was also experimentally verified in vitro.
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http://dx.doi.org/10.1016/j.fsi.2015.03.003DOI Listing
June 2015

Occurrence of zoonotic nematodes Pseudoterranova decipiens, Contracaecum osculatum and Anisakis simplex in cod (Gadus morhua) from the Baltic Sea.

Vet Parasitol 2014 Oct 6;205(3-4):581-7. Epub 2014 Sep 6.

Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark. Electronic address:

Baltic cod Gadus morhua (a total of total 224 specimens) captured east of the island of Bornholm in the southern Baltic Sea were subjected to a parasitological investigation between March 2013 and April 2014. Full artificial digestion of fillets from 188 cod and additional investigation of livers from 36 cod were performed. Cod or seal worm Pseudoterranova decipiens was recorded in musculature (prevalences up to 55% and intensities up to 56 worms per fish) associated with a negative correlation between worm intensity and condition factor. Liver worm Contracaecum osculatum (100% prevalence with intensities up to 320 worms per fish) in liver tissue were recorded but only a slight negative correlation between intensity and condition factor was noted. Seals act as final host for both worm species and the increased occurrence during recent years is associated with the increasing grey seal population in the area. Infection with Anisakis simplex (the herring or whale worm) in Baltic cod was found at a low level corresponding to previous studies.
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http://dx.doi.org/10.1016/j.vetpar.2014.08.027DOI Listing
October 2014

Characterization of serum amyloid A (SAA) in rainbow trout using a new monoclonal antibody.

Fish Shellfish Immunol 2014 Oct 19;40(2):648-58. Epub 2014 Aug 19.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark.

Serum amyloid A (SAA) is an integral part of the innate immune response in mammals and considered to be important during the acute phase response. The present study was undertaken to elucidate the role of SAA protein in the innate immune response of rainbow trout. A monoclonal antibody raised against a recombinant peptide of rainbow trout SAA was characterized using Western blot, dot blot, ELISA and immunohistochemistry. SAA association with high density lipoprotein (HDL) complicated band identification in Western blot, but delipidization of the SAA-HDL isolate highly increased the quality of reaction in the western blot. Rainbow trout fry (87 days post hatch) infected with Yersinia ruckeri showed a significant up-regulation of the SAA gene at 72 h post infection with an increase until 96 h post infection. Non-significant up-regulations were seen at earlier time points i.e. 4 and 24 h. The expression pattern of SAA significantly correlated to the immunohistochemical analysis of the infected fry. A weak staining was seen in liver tissue at 4 h post infection which increased in intensity during the course of infection i.e. 24, 72 and 96 h post infection.
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http://dx.doi.org/10.1016/j.fsi.2014.08.018DOI Listing
October 2014

Annual and spatial variability in endo- and ectoparasite infections of North Sea cod (Gadus morhua Linnaeus, 1758) larvae, post-larvae and juveniles.

Acta Parasitol 2014 Jun 15;59(2):284-93. Epub 2014 May 15.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, 1870, Frederiksberg C, Denmark,

A parasitological investigation was performed on a total of 5380 Atlantic cod larvae, post-larvae and small juveniles sampled from the North Sea during a period of five years. The copepod Caligus elongatus (Von Nordmann, 1832) and the nematode Hysterothylacium aduncum (Rudolphi, 1802) were found at a relatively high prevalence of infection (4.6% and 5.2%, respectively). The infection by both parasites showed annual and spatial variability. C. elongatus showed a higher prevalence in 1992 compared to the following years, whereas the prevalence of H. aduncum increased from 1992 to 2001.We observed a relation between parasite distribution and parameters such as latitude and water depth. Adult digeneans (Lecithaster gibbosus and Derogenes varicus) and larval cestodes were also found with lower infection rates. Since changes of infection levels coincided with increasing North Sea water temperature in the studied period, it is hypothesized that temperature may affect parasite population levels. However, it is likely that other environmental factors may contribute to the observed variations. Absence of infection intensities higher than one nematode per fish in small larvae and post-larvae suggests that host survival may be affected by a high infection pressure. The relatively high levels of infection in the younger stages of cod, and the annual/spatial variability of these infections should be considered in the understanding of the early life dynamics of the species.
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http://dx.doi.org/10.2478/s11686-014-0244-0DOI Listing
June 2014

Import of exotic and zoonotic trematodes (Heterophyidae: Centrocestus sp.) in Xiphophorus maculatus: implications for ornamental fish import control in Europe.

Acta Parasitol 2014 Jun 15;59(2):276-83. Epub 2014 May 15.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, 1870, Frederiksberg C, Denmark,

Ornamental fish, Xiphophorus maculatus, were imported from Singapore to Denmark for distribution to local aquarists. Importers observed lethargic and erratic swimming patterns among fish and forwarded a total of 30 fish for pathological examination to a university diagnostic service. All fish were diagnosed infected with encysted Centrocestus sp. metacercariae in gills (prevalence of 100% and mean intensity of 454.5 ± 161.9 parasites per fish). Metacercariae were identified by morphological and molecular methods. Cysts (mean length 163.3 ± 13.7 μm and mean width 113.3 ± 10.6 μm) contained a bent metacercaria with an X-shaped excretory bladder. PCR amplification of a rDNA region (5.8S rRNA gene, ITS-2, 28S rRNA gene) and subsequent sequencing confirmed the diagnosis. Metacercariae were found in gill filaments adjacent to the cartilage associated with cartilage hypertrophy, epithelial and mucous cell hyperplasia, clubbing and lamellar fusion. Host cell encapsulation of cysts comprised several layers of leucocytes, chondroblast-like and fibroblast-like cells. The observations raise concerns with regard to veterinary inspection and quarantine procedures. The zoonotic potential of these trematodes and a possible spread of the parasites in natural habitats in Europe should be regarded as a public health issue. So far, several cases of human infections have been reported only in Asia, but the potential intermediate host snail, Melanoides tuberculata, has been recorded in Germany. Accordingly, establishment of the parasite in Europe with climate changes should be considered a risk.
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http://dx.doi.org/10.2478/s11686-014-0237-zDOI Listing
June 2014

Increased Contracaecum osculatum infection in Baltic cod (Gadus morhua) livers (1982-2012) associated with increasing grey seal (Halichoerus gryphus) populations.

J Wildl Dis 2014 Jul 29;50(3):537-43. Epub 2014 Apr 29.

1  Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 7, Frederiksberg C, Denmark.

Grey seals (Halichoerus gryphus), the main final host of the gastric parasitic nematode Contracaecum osculatum in the Baltic, have recently recolonized the southwestern Baltic Sea. This colonization could lead to an increase in prevalence and intensity of third-stage larvae of C. osculatum in livers of Baltic cod (Gadus morhua), which serve as transport host for this helminth. We performed a parasitologic study of cod in spring 2012 and compared the results with previously unpublished data from 1982/1983. Additionally, grey seals were counted annually from 2000 to 2011 at three haul-out sites in the southwestern Baltic. Of 97 cod livers examined in the 1982/1983 survey, 22% harbored C. osculatum larvae, whereas 55.1% of the examined cod livers (n=185) were infected in 2012; the mean intensity and mean abundance increased from 4.3 and 0.9 to 20.2 and 11.1, respectively. Molecular identification (PCR) confirmed the identity of the larvae. The grey seal population increased markedly during the 12-yr period. We suggest that the elevated parasitism of cod livers is associated with the successful re-establishment of grey seals in the southwestern Baltic.
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http://dx.doi.org/10.7589/2012-12-320DOI Listing
July 2014

Expression of immune relevant genes in rainbow trout following exposure to live Anisakis simplex larvae.

Exp Parasitol 2013 Nov 16;135(3):564-9. Epub 2013 Sep 16.

Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 7, Frederiksberg C, Denmark. Electronic address:

Basic immune response mechanisms in vertebrates against helminths are still poorly understood. Fish-nematode models may prove valuable for elucidation of this question. In this study we orally challenged rainbow trout (Oncorhynchus mykiss) with larvae of Anisakis simplex (Nematoda: Anisakidae) and subsequently investigated the expression of 18 immune relevant genes in spleen and liver 1, 4 and 8days post infection (d.p.i.). Gene expression data were analysed with regard to the infection status of the challenged rainbow trout at the time of necropsy; "worms rejected" (÷worms), "worms present" (+worms) and a combined group consisting of samples pooled from both previous groups (÷/+worms). No significant regulation of cytokine genes was recorded but fish which had rejected worms up-regulated the CD4 gene (6.1-fold change, 8d.p.i.) in liver. The gene encoding CD8 was significantly down-regulated 24h post challenge in livers in fish still carrying worms (2.7-fold change) but not in the worm-free group. The immunoglobulin gene IgM was significantly down-regulated (2.9-fold change, 8d.p.i.) in liver samples from the +worms group. Complement factor C3 and precerebellin genes were significantly up-regulated twofold in liver samples from infected fish 4d.p.i. Significant up-regulation of the acute-phase protein SAA was observed in all three groups and in both tissues. To our knowledge, this is the first study to describe the expression of immune genes in a fish host challenged with live nematode larvae.
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http://dx.doi.org/10.1016/j.exppara.2013.09.011DOI Listing
November 2013

Insight from molecular, pathological, and immunohistochemical studies on cellular and humoral mechanisms responsible for vaccine-induced protection of rainbow trout against Yersinia ruckeri.

Clin Vaccine Immunol 2013 Oct 21;20(10):1623-41. Epub 2013 Aug 21.

Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg, Denmark.

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.
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http://dx.doi.org/10.1128/CVI.00404-13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3807201PMC
October 2013

Effects of excretory/secretory products from Anisakis simplex (Nematoda) on immune gene expression in rainbow trout (Oncorhynchus mykiss).

Fish Shellfish Immunol 2013 Sep 12;35(3):734-9. Epub 2013 Jun 12.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.

Excretory/secretory (ES) products are molecules produced by parasitic nematodes, including larval Anisakis simplex, a parasite occurring in numerous marine fish hosts. The effects of these substances on host physiology have not been fully described. The present work elucidates the influence of ES substances on the fish immune system by measuring immune gene expression in spleen and liver of rainbow trout (Oncorhynchus mykiss) injected intraperitoneally with ES products isolated from A. simplex third stage larvae. The overall gene expression profile of exposed fish showed a generalized down-regulation of the immune genes tested, suggesting a role of ES proteins in immunomodulation. We also tested the enzymatic activity of the ES proteins and found that lipase, esterase/lipase, valine and cysteine arylamidases, naphthol-AS-BI-phosphohydrolase and α-galactosidase activities were present in the ES solution. This type of hydrolytic enzyme activity may play a role in nematode penetration of host tissue. In addition, based on the notion that A. simplex ES products may have an immune-depressive effect (by minimizing immune gene expression) it could also be suggested that worm enzymes directly target host immune molecules which would add to a decreased host immune response and increased worm survival.
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http://dx.doi.org/10.1016/j.fsi.2013.06.007DOI Listing
September 2013

Comparative infectivity of three larval nematode species in three different salmonids.

Parasitol Res 2013 Aug 4;112(8):2997-3004. Epub 2013 Jun 4.

Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Frederiksberg, Denmark.

Host specificity of parasites may depend both on ecological and physiological factors. Basic descriptions of the susceptibility/resistance of fish to specific nematodes are needed in order to reveal mechanisms in the host-parasite relation. Rainbow trout (Oncorhynchus mykiss), brown trout (Salmo trutta), and Atlantic salmon (Salmo salar) were orally infected with larval stages of three different anisakid nematodes-Hysterothylacium aduncum, Contracaecum osculatum, and Anisakis simplex-and parasite survival and location was subsequently recorded for up to 14 days post infection (dpi). H. aduncum was most prevalent and numerous in brown trout 2 dpi, but a large proportion of the worms were recovered dead. No tissue penetration was observed. Rainbow trout exhibited the highest susceptibility to C. osculatum larvae at 2, 7, and 14 dpi. Mean intensities and mean abundances were lower in brown trout and salmon at all time points. The pyloric cecum was penetrated in rainbow trout on two occasions. A. simplex larvae established more successfully in salmon compared to rainbow trout; brown trout exhibited the highest natural resistance. Mean intensity and mean abundance was highest in rainbow trout at 2 and 7 dpi, but not after 14 days. A range of tissues, including muscle and liver, were found penetrated by larvae, but the pyloric ceca were the preferred microhabitat for Anisakis in both rainbow trout and salmon. It can be concluded that closely related salmonids differ in susceptibility towards different anisakid larvae and these parasites select different microhabitats in the hosts. The physiological basis for this specificity is discussed.
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http://dx.doi.org/10.1007/s00436-013-3472-6DOI Listing
August 2013

Myxobolus groenlandicus n. sp. (Myxozoa) distorting skeletal structures and musculature of Greenland halibut Reinhardtius hippoglossoides (Teleostei: Pleuronectidae).

Dis Aquat Organ 2012 Mar;98(2):133-41

Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Denmark.

A specimen of Greenland halibut Reinhardtius hippoglossoides (Walbaum, 1792) caught on the west coast of Greenland (Qasigiannguit) was found to possess serious pathological changes in the body musculature. A series of cartilaginous cylindrical structures organized symmetrically at the position of the proximal pterygiophores had changed the musculature and produced irreversible distortions (cavities and holes) in the fillet of the processed fish, leaving it with no value for the industry. Histopathological investigation showed that these structures consisted of hypertrophic cartilage containing numerous myxospore-producing plasmodia. Morphometric and molecular analyses of the parasites showed that both spore morphology and rDNA sequences complied with characteristics of the genus Myxobolus, but no full affiliation with a known species could be found. The parasite is a previously undescribed species, and the name Myxobolus groenlandicus n. sp. is assigned to this new myxobolid.
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http://dx.doi.org/10.3354/dao02437DOI Listing
March 2012

Differential immune response of rainbow trout (Oncorhynchus mykiss) at early developmental stages (larvae and fry) against the bacterial pathogen Yersinia ruckeri.

Dev Comp Immunol 2012 Feb 10;36(2):463-74. Epub 2011 Sep 10.

University of Copenhagen, Faculty of Life Sciences, Department of Veterinary Disease Biology, Frederiksberg C, Denmark.

Innate immune factors play a crucial role in survival of young fish especially during early stages of life when adaptive immunity is not fully developed. In the present study, we investigated the immune response of rainbow trout (Oncorhynchus mykiss) larvae and fry at an early stage of development. We exposed 17 and 87° days post hatch larvae and fry (152 and 1118 degree days post hatch; avg. wt. 70 and 770 mg, respectively) to the bacterial pathogen, Yersinia ruckeri for 4h by bath challenge. Samples were taken at 4, 24, 72 and 96 h post exposure for qPCR and immunohistochemical analyses to elucidate the immune response mounted by these young fish. Larvae showed no mortality although infected larvae at 48 h post exposure showed hyperaemia in the mouth region and inflammation on the dorsal side of the body. Gene expression studies showed an up-regulation of iNOS and IL-22 in infected larvae 24h post exposure but most of the investigated genes did not show any difference between infected and uninfected larvae. Immunohistochemical studies demonstrated a high expression of IgT molecules in gills and CD8 positive cells in thymus of both infected and uninfected larvae. Infection of rainbow trout fry with Y. ruckeri, in contrast, induced a cumulative mortality of 74%. A high expression of cytokines (IL-1β, TNF-α, IL-22, IL-8 and IL-10), acute phase proteins (SAA, hepcidin, transferrin and precerebellin), complement factors (C3, C5 and factor B), antimicrobial peptide (cathelicidin-2) and iNOS was found in infected fry when compared to the uninfected control. IgT molecules and mannose binding lectins in gills of both infected and uninfected fry were detected by immunohistochemistry. The study indicated that early life stages (yolk-sac larvae), merely up-regulate a few genes and suggests a limited capacity of larvae to mount an immune response by gene regulation at the transcriptional level. Based on the observed clearance of bacteria and lack of mortality it could be speculated that larvae may be covered by protective shield of different immune factors providing protection against broad range of pathogens. However, the increased susceptibility of older fry suggests that Y. ruckeri may utilize some of the immune elements to enter the naive fish. The up-regulation of iNOS and IL-22 in the infected larvae implicates an important role of these molecules in immune response at early developmental stages. A dense covering of surfaces of gill filaments by IgT antibody in the young fish suggest a role of this antibody as innate immune factor at early developmental stages.
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http://dx.doi.org/10.1016/j.dci.2011.08.014DOI Listing
February 2012

Cellular and humoral factors involved in the response of rainbow trout gills to Ichthyophthirius multifiliis infections: molecular and immunohistochemical studies.

Fish Shellfish Immunol 2011 Mar 25;30(3):859-69. Epub 2011 Jan 25.

Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Stigbøjlen 7, DK-1870 Frederiksberg C, Denmark.

The parasitic ciliate Ichthyophthirius multifiliis infecting skin, fins and gills of fish induces a protective immune response in rainbow trout (Oncorhynchus mykiss) surviving the infection and a similar protection can be conferred by i.p. injection of live theronts. A combined molecular and immunohistochemical approach has been used in this work for pinpointing cellular and humoral immune factors in gill tissue involved in the response and indicating interactions between the systemic and local responses. Fish were immunized by intra-peritoneal injection of live I. multifiliis theronts, control fish were injected with PBS and subgroups were treated with the immuno-suppressant hydrocortisone before fish were challenged with live theronts. Significant up-regulations of genes encoding IgM, IgT, C3, SAA, IL-8, IL-22 and IFN-γ were induced by immunization and challenge. Hydrocortisone treatment had a significant down-regulating effect on genes incoding IgT, IgM, CD4, CD8, IFN-γ, IL-8 and IL-22 in all groups. Immunohistochemistry, using monoclonal antibodies to detect cellular markers, demonstrated active involvement of CD8, MHC II, IgT and IgM positive cells in gill tissue. Putative T-cells (CD8 positive cells) were detected in the intraepithelial lymphoid tissue located at the base of gill filaments and in hyperplastic gill tissue but following infection a clear efflux of these cells was detected. MHC II positive cells were distributed across the gill filaments and accumulated in hyperplastic tissue but hydrocortisone treatment affected their density negatively in both immunized and non-immunized fish. IgT positive cells were present in the epithelial lining of the gill lamellae (suggesting a primary role of this protein in the mucosal defence against the ciliate) whereas IgM positive cells were found only in gill arterioles and the lamellar capillaries. The present work indicates an intensive activity and specialized function of immune cells (B-cells, T-cells and macrophages) and humoral elements such as immunoglobulins IgT and IgM which are orchestrated by cytokines in gill tissue reacting against I. multifiliis.
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http://dx.doi.org/10.1016/j.fsi.2011.01.010DOI Listing
March 2011

PAMP induced expression of immune relevant genes in head kidney leukocytes of rainbow trout (Oncorhynchus mykiss).

Dev Comp Immunol 2011 Apr 13;35(4):476-82. Epub 2010 Dec 13.

Section of Biomedicine, Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark.

Host immune responses elicited by invading pathogens depend on recognition of the pathogen by specific receptors present on phagocytic cells. However, the reactions to viral, bacterial, parasitic and fungal pathogens vary according to the pathogen-associated molecular patterns (PAMPs) on the surface of the invader. Phagocytic cells are known to initiate a respiratory burst following an exposure to the pathogen, but the underlying and associated specific elements are poorly elucidated in fish. The present study describes the differential response of head kidney leukocytes from rainbow trout (Oncorhynchus mykiss) to different PAMPs mimicking viral (poly I:C), bacterial (flagellin and LPS) and fungal infections (zymosan and β-glucan). Transcript of cytokines related to inflammation (IL-1β, IL-6, IL-10 and TNF-α) was highly up-regulated following LPS exposure whereas flagellin or poly I:C induced merely moderate reactions. In contrast, IFN-γ expression was significantly higher in the poly I:C stimulated group compared to the LPS group. When head kidney cells were exposed to zymosan or β-glucan, genes encoding IL-1β, TNF-α, IL-6 and IL-10 became up-regulated. Their level of up-regulation was comparable to LPS but the kinetics differed. In particular, TNF-α induction was considerably slower when stimulated with zymosan or β-glucan. The gene encoding the COX-2 enzyme, a central element during initiation of inflammatory reactions, was significantly higher in stimulated cells although a depressing effect of high concentrations of LPS and zymosan became evident after 4h exposure. This study suggests that rainbow trout leukocytes respond differently to viral, bacterial and fungal PAMPs, which may reflect activation of specific signaling cascades eventually leading to activation of different immune effector molecules.
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http://dx.doi.org/10.1016/j.dci.2010.12.001DOI Listing
April 2011
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