Hymenopteran parasitoids reared from European gall midges (Diptera, Cecidomyiidae).

We report the results of investigations 2010 through 2023 of hymenopteran parasitoids associated with gall midges in Europe. A total of 242 collections of gall midges were made, from each of which one to several parasitoid species emerged, resulting in ca. 200 recorded parasitoid species and 267 host-parasitoid interaction records. The parasitoid families involved were Eulophidae (63 species), Platygastridae (56 species), Torymidae (34 species), Pteromalidae (31 species), Ceraphronidae (5 species), Eupelmidae (4 species), Eurytomidae (2 species) and Encyrtidae (1 species). As many as 159 interactions are reported for the first time, significantly enlarging our knowledge of gall midge - parasitoid interactions on the species level. Even more interesting, 51 host records are for parasitoid species for which no host was previously known. Similarly, 28 species of gall midge are reported as host to named parasitoids for the first time. Additionally, 91 parasitoid records were the first for the country in question. Differences between the rearing methods applied and their suitability for recording species with contrasting life histories, are discussed.

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

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 We performed a parasitologic study of cod in spring 2012 and compared the results with previously unpublished data from 1982/1983.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.

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

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.

Occurrence of Diplostomum pseudospathaceum Niewiadomska, 1984 and D. mergi Dubois, 1932 (Digenea: Diplostomidae) in Danish freshwater snails: ecological and molecular data.

Freshwater pulmonate snails from three locations in Lake Furesø north of Copenhagen were screened for infection with furcocercariae (by shedding in the laboratory) and recovered parasite larvae were diagnosed by molecular methods (by performing PCR of rDNA and sequencing the internal transcribed spacer [ITS] region). Overall prevalence of infection in snails was 2%. Recovered cercariae from Lymnaea stagnalis (Linnaeus) were diagnosed as Diplostomum pseudospathaceum Niewiadomska, 1984 (prevalence 4%) and cercariae from Radix balthica (Linnaeus) as D. mergi (Dubois, 1932) (prevalence 2%). Pathogen-free rainbow trout were then exposed to isolated cercariae and infection success and site location of metacercariae in these fish were determined. Infection experiments confirmed that both species could infect rainbow trout with the eye lens as infection site for the metacercarial stage although infection success differed. Combination of molecular and biological assays may contribute to improvement of our knowledge on diagnosis, distribution and biology of diplostomids in fish.

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

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.

Microhabitat preference of Anisakis simplex in three salmonid species: immunological implications.

Three salmonid fish species, Oncorhynchus mykiss, Salmo salar and Salmo trutta, were infected experimentally with the parasitic nematode Anisakis simplex (A. simplex) and the difference between in vivo behaviour of the nematode in the three fish species was investigated. Infection success rate differed between species. S. salar (Baltic salmon) showed the highest number of successfully established nematodes, whereas S. trutta (brown trout) and O. mykiss (rainbow trout) had a higher natural resistance. Microhabitat selection of nematodes differed according to fish species. In brown trout, A. simplex larvae were attached to the digestive tract (stomach, pyloric caeca, intestine), while the majority of larvae found in rainbow trout were located between the pyloric caeca. In Baltic salmon, nematodes were dispersed in and on spleen, head kidney, liver, swim bladder and musculature. Encapsulation and inflammatory cellular reactions differed accordingly. Histopathological and immunohistochemical studies using monoclonal antibodies raised against salmonid IgM, CD8 and MHCII were performed to detect the presence of immune cells around the infecting nematodes. None of the three fish species showed positive reactions for IgM-bearing cells in the inflammatory tissue connected with nematodes. CD8+ cells were detected in all three species and MHCII-bearing cells were found associated with encapsulated A. simplex in rainbow trout and brown trout, but not in Baltic salmon. Physiological, immunological and pathological implications of microhabitat differences are discussed.