The involvement of Pseudoterranova decipiens fish infestation on the shelf-life of fresh Atlantic cod (Gadus morhua) fillet.

Zoonotic nematodes of the family Anisakidae are highly common in many marine fish species, which act as paratenic hosts for the third larval stage. In the fish, these parasites may migrate from the fish's gastro-intestinal tract (GI-tract) further to the coelomic cavity and muscles, making them a possible contamination source of bacteria they carry on their cuticle and in their GI-tract. A previous study revealed no apparent effect of Anisakis simplex on spoilage of fish, but the equally common anisakid species Pseudoterranova decipiens has a larger body surface potentially increasing the bacterial load brought into the fish muscle upon migration. As the presence of shelf-life reducing spoilage bacteria in the microbiome of this anisakid species has been demonstrated, the objective of the present study was to assess the potential shelf-life reducing effect of P. decipiens in fresh fish fillets stored in a domestic refrigerator. Atlantic cod was used as a model since members of the cod family are the third most consumed marine fish globally and it has the highest prevalence of P. decipiens infections. Infected and non-infected codfish fillet portions were collected and microbiologically analyzed at day 0 and day 4 of storage in a domestic fridge. Three isolation media were used to enhance maximum bacterial recovery and isolates were identified using MALDI-TOF MS and 16S rRNA gene sequencing. In parallel to the microbiological examination, sensory analysis was performed daily on the cod fillets to evaluate the freshness of the fish. Results revealed the presence of typical spoilage bacteria (e.g., Pseudomonas sp., Photobacterium sp.) in all fish, but based on the total viable counts, total H2S-producing bacteria, and sensory analysis, there were no objective indications to assume an increased fish spoilage rate by the presence and migration P. decipiens. Additionally, a beta-diversity comparison revealed no significant differences in microbiota composition between infected and non-infected fish parts, though individual heterogeneity in microbiome composition among Atlantic codfish individuals was found. As total viable counts did, however, exceed the guideline limits for fresh fish, further research should now focus on the role of the candling step as a potential source of post-harvest contamination. As such, anisakid infection might still accelerate fish spoilage, though now in an indirect way.

Seals, fish, humans and parasites in the Baltic: ecology, evolution and history.

Evolutionary and ecological processes affecting the interactions between hosts and parasites in the aquatic environment are at display in the Baltic Sea, a young and ecologically unstable marine ecosystem, where fluctuating abiotic and biotic factors affect the parasitofauna in fish. The dynamic infections of Baltic cod, a subpopulation of the Atlantic cod (Gadus morhua Linnaeus), with third stage anisakid nematode larvae of Pseudoterranova decipiens (Krabbe, 1878) and Contracaecum osculatum (Rudolphi, 1802) have increased following a significant increase of the Baltic grey seal Halichoerus grypus (Fabricius) population in the region. Cod serves as a paratenic host and marine mammals, pinnipeds, are definitive hosts releasing parasite eggs, with faeces, to the marine environment, where embryonation and hatching of the third stage larva take place. The parasite has no obligate intermediate hosts, but various invertebrates, smaller fish and cod act as paratenic hosts transmitting the infection to the seal. Contracaecum osculatum has an impact on the physiological performance of the cod, which optimises transmission of the larva from fish to seal. Thus, a muscle mass decrease of nearly 50% may result from heavy C. osculatum infections, probably amplified by a restricted food availability. The muscle atrophy is likely to reduce the escape reactions of the fish when meeting a foraging seal. In certain regions, where fish and seals are restricted in their migration patterns, such as the semi-enclosed Baltic Sea, the predation may contribute to a severe cod stock depletion. The parasites are zoonotic and represent a human health risk, when consumers ingest insufficiently heat- or freeze-treated infected products. Marked infections of the cod were previously reported during periods with elevated seal populations (late 19th and middle 20th century) and various scenarios for management of risk factors are evaluated in an evolutionary context.

Unraveling the microbiota of the fish parasite Pseudoterranova decipiens in codfish (Gadus morhua) reveals a fish-related bacterial community.

Anisakidae, mainly represented by the species Anisakis simplex and Pseudoterranova decipiens, are one of the most commonly zoonotic nematodes present in marine fish species. Apart from public health risks directly linked to the parasite itself, little is known on the effects of the migrating nematodes on the hygienic quality of the fish fillet due to bacteria it carries. In the present study, the cultivated bacterial community on and in individual P. decipiens larvae deriving from codfish is reported. Four isolation media were included and evaluated to increase the bacterial diversity isolated, and identification of the bacterial growth was performed by a combination of Matrix-Assisted Laser Desorption Ionization Time-Of-Flight mass spectrometry and 16S rRNA gene sequencing. Results revealed that the microbiota of P. decipiens larvae comprises both potential spoilage bacteria and human opportunistic pathogens, and that a combined isolation on the general isolation medium tryptone soy agar and a medium supplemented with artificial seawater resulted in the highest bacterial recovery in terms of diversity and enumeration. Dissimilarity analysis also revealed similar, though unique, bacterial communities between nematodes originating from the same fish suggesting that anisakid microbiota compositions are reflections of the microbial assemblages in the fish host as an individual, and that the gut microbiome is diverse within gadoid fish species originating from the same geographical habitat. Future research should, based on the results in the present study, further elaborate on the comparison of the bacterial communities of both the larva and the codfish from which it was isolated, and, explore the extrapolation potential towards other fish and nematode species. Also, the actual degree of risk beyond the simple presence of the parasite due to carriage of opportunistic bacteria should be examined, as well as the nematode's true effect on spoilage.

In situ evidence of the role of Crangon crangon in infection of cod Gadus morhua with nematode parasite Hysterothylacium aduncum in the Baltic Sea.

Cod was one of the most important fish species in the Baltic Sea, but its condition is deteriorating for several reasons, including an increasing parasite burden. The aim of this study was to determine the source of infection of Baltic cod with parasites by examination of invertebrates found in situ in the cod stomach. A total of 1681 cod were sampled during four research cruises in the southern Baltic Sea in 2012, 2013 and 2014 and the composition of their diet was analysed. Each prey item from cod stomach was identified to the lowest possible taxonomic level and a parasitological analysis of all invertebrates collected was performed. Crangon crangon, Saduria entomon and Mysis mixta were the most commonly represented invertebrates among food items. Hysterothylacium aduncum was found only in C. crangon. This host­parasite system is reported here for the first time in situ in the stomach of cod from the Baltic Sea, confirming the role of C. crangon in cod infection with H. aduncum.

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

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.

Contracaecum osculatum (sensu lato) infection of Gadus morhua in the Baltic Sea: inter- and intraspecific interactions.

The subpopulation of Atlantic cod, Gadus morhua, in the eastern part of the Baltic Sea has experienced a significant increase in infections with anisakid nematode larvae of the species Contracaecum osculatum sensu lato (s.l.) since the year 2000. The life cycle of the parasite includes seals and especially the grey seal, Halichoerus grypus, as final hosts, carrying the adult nematodes in the stomach, crustaceans (copepods, amphipods) as first intermediate hosts and various fish species (clupeids, sandeel) including cod as second intermediate/paratenic hosts. Cod with a body length below 28 cm are generally non-infected but experience increasing infection levels when they switch to a piscine diet (infected intermediate/paratenic hosts). We present an overall frequency distribution analysis of worms in 166 cod (body length 30-49 cm) collected in the spawning area over the last 5 years. It shows a fit to the negative binomial distribution, a prevalence of infection of 89.8%, a mean intensity of 29.3 parasites per fish (range 1-377) and a variance/mean ratio of 59.2 (≫1), indicating overdispersion. We present measurements of the adult Contracaecum osculatum (s.l.) specimens in the seal stomach and show that the parasites reach a maximum length of 6.6 cm (females) and 5.8 cm (males). L3s in sprat have a total length from 1to 11 mm whereas the larvae in cod liver are 3-27 mm. A decreasing mean worm length associated with high worm densities in cod (number of nematodes per liver) was recorded. Possible explanations might include timing of feeding on infected intermediate/paratenic hosts, intraspecific competition (crowding) between larvae in cod and host responses (indicated by a significant antibody production in cod against C. osculatum (s.l.) antigens). A significant negative correlation between infection intensity and muscle mass of cod was found, suggesting parasite-induced down-regulation of growth factors in cod.

Baltic cod endohelminths reflect recent ecological changes.

We suggest helminthological investigations of cod as a supplement to traditional biological and hydrographical methods for elucidation of ecological changes in the Baltic Sea. It is under discussion if oxygen deficit or seal abundance should explain the present critical situation of Baltic cod. A comparative investigation of endoparasitic helminths in Baltic cod (Gadus morhua), captured in the same marine habitat with an interval of 35 years (1983/2018) recorded 11 species of helminths comprising trematodes (Hemiurus luehei, Podocotyle atomon, Lepidapedon elongatum), nematodes (Contracaecum osculatum, Hysterothylacium aduncum, Capillaria gracilis, Cucullanus cirratus), cestodes (Bothriocephalus sp.) and acanthocephalans (Echinorhynchus gadi, Pomphorhynchus laevis, Corynosoma semerme). Significant prevalence and intensity increases were recorded for third-stage larvae of the nematode C. osculatum (liver location) and larvae of C. semerme (encapsulated in viscera). Both parasite species use grey seal as their final host, indicating the recent expansion of the Baltic seal population. A lower E. gadi intensity and an increased prevalence of L. elongatum of small cod (31-40 cm body length) suggest a lowered intake of amphipods (intermediate host) and elevated ingestion of polychaetes, respectively, but no significant changes were seen for other helminths.

Local immune depression in Baltic cod (Gadus morhua) liver infected with Contracaecum osculatum.

Third-stage larvae of the anisakid nematode Contracaecum osculatum infecting cod (Gadus morhua) liver elicit a host immune response involving both innate and adaptive factors, but the reactions differ between liver and spleen. Inflammatory reactions occur in both liver and spleen, but a series of immune effector genes are downregulated in liver infected with nematodes whereas these genes in spleen from the same fish are upregulated. A series of novel primer and probe sets targeting cod immune responses were developed and applied in a real-time quantitative polymerase chain reaction set-up to measure the expression of immune-relevant genes in liver and spleen of infected and uninfected cod. In infected liver, 12 of 23 genes were regulated. Genes encoding cytokines associated with inflammatory reactions (IL-1ß, IL-6, IL-8) were significantly upregulated, whereas genes encoding effector molecules, assisting the elimination of pathogens, C-reactive protein (CRP)-PII, hepcidin, lysozyme G1, lysozyme G2, C3 and IgDm, were significantly downregulated. The number of downregulated genes increased with the parasite burden. In spleen, 14 of 23 immune genes showed significant regulation and nine of these were upregulated, including genes encoding CRPI, CRPII, C3, hepcidin and transferrin. The general gene expression level was higher in spleen compared to liver, and although inflammation was induced in nematode-infected liver, the effector molecule genes were depressed, which suggests a worm-induced immune suppression locally in the liver.

First evidence of the presence of Anisakis simplex in Crangon crangon and Contracaecum osculatum in Gammarus sp. by in situ examination of the stomach contents of cod (Gadus morhua) from the southern Baltic Sea.

Cod (Gadus morhua), an important fish species in the Baltic Sea, is the paratenic host for many parasite species, including the zoonotic nematodes, Anisakis sp. and Contracaecum osculatum. We aimed to identify which invertebrate species (found in situ in the fish stomach) are responsible for infection of cod with zoonotic nematodes. We found that Crangon crangon and Gammarus sp., both invertebrate prey species of cod, were infected with Anisakis simplex and C. osculatum, respectively. These host-parasite systems are reported here for the first time, implicating C. crangon and Gammarus sp. as sources of infection of Baltic cod with zoonotic nematodes.

How effective is freezing at killing Anisakis simplex, Pseudoterranova krabbei, and P. decipiens larvae? An experimental evaluation of time-temperature conditions.

The consumption of raw or inadequately cooked marine fish can lead to several disorders caused by the ingestion of viable anisakid nematodes. Although anisakid larvae can be killed by subzero temperatures, making freezing an important control measure for this potential health hazard, these parasites can survive freezing under some conditions. Therefore, the aim of the present study was to experimentally evaluate the time-temperature conditions needed to kill Anisakis simplex and Pseudoterranova spp. The effectiveness of freezing was tested on two species of fish: cod, Gadus morhua from the North Atlantic, and herring, Clupea harengus membras from the southern Baltic Sea. Samples, which comprised skinless fillets of cod (n = 40) with visible parasites and whole herring (n = 240), were separately frozen at - 15, - 18, or - 20 °C for 24 h, or at - 20 °C for 48 h in the single-compressor freezer and at - 20, - 25, or - 35 °C for 24 h in the double-compressor freezer. After thawing, parasites were stained with malachite green and examined under the microscope for viability. All A. simplex and Pseudoterranova spp. larvae in cod fillets died at a temperature of - 15 °C or lower. However, freezing did not kill all the A. simplex larvae in whole herring: spontaneous movement of these parasites was observed in samples stored in the single-compressor freezer at - 15, - 18, and - 20 °C over 24 h. Our results demonstrate that the freezing procedure must consider both the capability of the freezing device and the nature of the fish product to ensure consumer safety.

A Novel Method for Predicting Anisakid Nematode Infection of Atlantic Cod Using Rough Set Theory.

Atlantic cod ( Gadus morhua L.) is one of the most important fish species in the fisheries industries of many countries; however, these fish are often infected with parasites. The detection of pathogenic larval nematodes is usually performed in fish processing facilities by visual examination using candling or by digesting muscles in artificial digestive juices, but these methods are both time and labor intensive. This article presents an innovative approach to the analysis of cod parasites from both the Atlantic and Baltic Sea areas through the application of rough set theory, one of the methods of artificial intelligence, for the prediction of food safety in a food production chain. The parasitological examinations were performed focusing on nematode larvae pathogenic to humans, e.g., Anisakis simplex, Contracaecum osculatum, and Pseudoterranova decipiens. The analysis allowed identification of protocols with which it is possible to make preliminary estimates of the quantity and quality of parasites found in cod catches before detailed analyses are performed. The results indicate that the method used can be an effective analytical tool for these types of data. To achieve this goal, a database is needed that contains the patterns intensity of parasite infections and the conditions of commercial fish species in different localities in their distributions.

Saduria entomon infected with Hysterothylacium aduncum found in situ in the stomach of cod (Gadus morhua) from the Baltic Sea.

The parasite fauna of cod (Gadus morhus) is well described, but the life cycles of Baltic cod parasites are known only in general terms. Invertebrates commonly found in the stomach of cod are recognized as intermediate hosts in the life cycles of nematodes or acanthocephalans. The aim of this study was to determine the source of infection of Baltic cod with parasites found in situ in invertebrates present in the cod stomach. Our results indicate that Saduria entomon is both a source of infection of Baltic cod with parasites and an intermediate host in the life cycle of Hysterothylacium aduncum in the Baltic Sea.

Contracaecum osculatum and other anisakid nematodes in grey seals and cod in the Baltic Sea: molecular and ecological links.

Populations of grey seals (Halichoerus grypus), sprats (Sprattus sprattus) and cod (Gadus morhua) in the Baltic Sea are relatively stationary. The present work, applying classical and molecular helminthological techniques, documents that seals and cod also share a common parasite, the anisakid nematode Contracaecum osculatum, which uses seals as the final host and fish as transport hosts. Sequencing mitochondrial genes (COX1 and COX2) in adult worms from seals and third-stage larvae from livers of Baltic fish (sprats and cod), showed that all gene variants occur in both seals and fish. Other anisakid nematodes Pseudoterranova decipiens and Anisakis simplex are also found in both seals and cod in the Baltic Sea, but at much lower rates. The Baltic grey seal population was left at a critically low level (comprising a few hundred individuals) during the latter part of the 20th century, but since the year 2000 a marked increase in the population has been observed, reaching more than 40,000 individuals at present. Ecological consequences of the increased seal abundance may result from increased predation on fish stocks, but recent evidence also points to the influence of elevated parasitism on fish performance. Contracaecum osculatum larvae preferentially infect the liver of Baltic cod, considered a vital organ of the host. Whereas low prevalences and intensities in cod were reported during the 1980s and 1990s, the present study documents 100% prevalence and a mean intensity of above 80 worms per fish. Recent studies have also indicated the zoonotic potential of C. osculatum larvae in fish, following the consumption of raw or under-cooked fish. Therefore the present work discusses the impact of parasitism on the cod stock and the increasing risk for consumer health, and lists possible solutions for control.

Extrusion of Contracaecum osculatum nematode larvae from the liver of cod (Gadus morhua).

Baltic cod livers have during recent years been found increasingly and heavily infected with third-stage larvae of Contracaecum osculatum. The infections are associated with an increasing population of grey seals which are final hosts for the parasite. Heavy worm burdens challenge utilization and safety of the fish liver products, and technological solutions for removal of worms are highly needed. We investigated the attachment of the worm larvae in liver tissue by use of histochemical techniques and found that the cod host encapsulates the worm larvae in layers of host cells (macrophages, fibroblasts) supported by enclosures of collagen and calcium. A series of incubation techniques, applying compounds targeting molecules in the capsule, were then tested for their effect to induce worm escape/release reactions. Full digestion solutions comprising pepsin, NaCl, HCl and water induced a fast escape of more than 60% of the worm larvae within 20 min and gave full release within 65 min but the liver tissue became highly dispersed. HCl alone, in concentrations of 48 and 72 mM, triggered a corresponding release of worm larvae with minor effect on liver integrity. A lower HCl concentration of 24 mM resulted in 80% release within 35 min. Water and physiological saline had no effect on worm release, and 1% pepsin in water elicited merely a weak escape reaction. In addition to the direct effect of acid on worm behaviour it is hypothesised that the acid effect on calcium carbonate in the encapsulation, with subsequent release of reaction products, may contribute to activation of C. osculatum larvae and induce escape reactions. Short-term pretreatment of infected cod liver and possibly other infected fish products, using low acid concentrations is suggested as part of a technological solution for worm clearance as low acid concentrations had limited macroscopic effect on liver integrity within 35 min.

Immunomodulatory effects of excretory/secretory compounds from Contracaecum osculatum larvae in a zebrafish inflammation model.

Excretory/secretory (ES) compounds isolated from third-stage larvae of the anisakid nematode Contracaecum osculatum parasitizing liver of Baltic cod were investigated for effects on immune gene expression in a zebrafish LPS-induced inflammation model. ES products containing a series of proteins, of which some had enzymatic activity, were injected solely or with LPS. ES proteins alone induced up-regulation of a number of immune-related genes, but generally to a lower degree compared to LPS. When co-injected with LPS, the worm products exacerbated merely expression of five genes affecting Th1, Th2, Th17 and innate responses compared to the LPS-injected group. However, the level of overexpression decreased in an inverse dose-dependent manner. The immune regulating action of C. osculatum ES products is interpreted as an important evolutionary ability of larval parasites in the transport host which makes it less susceptible to host immune responses whereby the probability of reaching the final host is increased.

Host condition and accumulation of metals by acanthocephalan parasite Echinorhynchus gadi in cod Gadus morhua from the southern Baltic Sea.

In this study, we analyzed the relationship between concentration of metals in the host-parasite system (cod - acanthocephalan Echinorhynchus gadi) and Fulton's condition factor (FCF) of the host. The relationship between metal (Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Pb, Sr, Zn) concentrations in E. gadi and cod tissues was expressed as a bioconcentration factor (BCF), the ratio of the concentration in the parasite tissue to that in host tissues. Acanthocephalans accumulated mainly toxic metals (Cd, Pb), as well as Sr, Ca, Na. Cadmium showed the highest bioconcentration in parasites (BCF >200) compared to fish muscle. Significant negative correlation was detected between FCF and the concentration of Cd and Hg in cod liver. In contrast, FCF was positively correlated with the concentration of Hg in acanthocephalan tissues.

Host size-dependent anisakid infection in Baltic cod Gadus morhua associated with differential food preferences.

A significant increase in the infection level of Baltic cod Gadus morhua with the anisakid nematode larvae Contracaecum osculatum and Pseudoterranova decipiens has been recorded during recent years due to the expanding local population of grey seals Halichoerus grypus, which act as final hosts for these parasites. Here, we report from an investigation of 368 cod (total length [TL] 6-49 cm; caught in ICES Subdivision 25) that the infection level of juvenile cod (TL 6-30 cm) with larvae of C. osculatum and P. decipiens is absent or very low, whereas it increases drastically in larger cod (TL 31-48 cm). A third nematode Hysterothylacium aduncum was rarely found. The study indicates that the prey animals for large cod act as transport hosts for the parasite larvae. Analyses of stomach contents of cod caught in the same area (2007-2014) showed that small benthic organisms (including polychaetes Harmothoë sarsi) are preferred food items by small cod, the isopod Saduria entomon is taken by all size classes, and sprat Sprattus sprattus are common prey items for cod larger than 30 cm. Parasitological investigations (microscopic and molecular analyses) of H. sarsi (100 specimens) and S. entomon (40 specimens) did not reveal infection in these invertebrates, but 11.6% of sprat (265 specimens examined) was shown to be infected with 1-8 C. osculatum third stage larvae per fish. Analyses of sprat stomach contents confirmed that copepods and cladocerans are the main food items of sprat. These observations suggest that the C. osculatum life cycle in the Baltic Sea includes grey seals as final hosts, sprat as the first transport host and cod as second transport host. It may be speculated that sprat obtain infection by feeding on copepods and/or cladocerans, which could serve as the first intermediate hosts. One cannot exclude the possibility that the size-dependent C. osculatum infection of cod may contribute (indirectly or directly) to the differential mortality of larger cod (>38 cm) compared to smaller cod (<30 cm) recently recorded in the Baltic cod population.

Measuring abnormal movements in free-swimming fish with accelerometers: implications for quantifying tag and parasite load.

Animal-borne data loggers allow movement, associated behaviours and energy expenditure in fish to be quantified without direct observations. As with any tagging, tags that are attached externally may adversely affect fish behaviour, swimming efficiency and survival. We report on free-swimming wild Atlantic cod (Gadus morhua) held in a large mesocosm that exhibited distinctly aberrant rotational swimming (scouring) when externally tagged with accelerometer data loggers. To quantify the phenomenon, the cod were tagged with two sizes of loggers (18 and 6 g; <2% body mass) that measured tri-axial acceleration at 50 Hz. An automated algorithm, based on body angular rotation, was designed to extract the scouring movements from the acceleration signal (98% accuracy). The algorithm also identified the frequency pattern and associated energy expenditure of scouring in relation to tag load (% body weight). The average per cent time spent scouring (5%) was independent of tag load. The vector of the dynamic body acceleration (VeDBA), used as a proxy for energy expenditure, increased with tag load (r(2)=0.51), and suggests that fish with large tags spent more energy when scouring than fish with small tags. The information allowed us to determine potential detrimental effects of an external tag on fish behaviour and how these effects may be mitigated by tag size. The algorithm can potentially identify similar rotational movements associated with spawning, courtship, feeding and parasite-load shedding in the wild. The results infer a more careful interpretation of data derived from external tags and the careful consideration of tag type, drag, buoyancy and placement, as well as animal buoyancy and species.

Spatial ecology of coastal Atlantic cod Gadus morhua associated with parasite load.

Acoustic tags and receivers were used to investigate the spatial ecology of coastal Atlantic cod Gadus morhua (n = 32, mean fork length: 50 cm, range: 33-80 cm) on the Norwegian Skagerrak coast in 2012. Monthly home ranges (HR), swimming activity and depth use varied considerably among individuals and through the months of June, July and August. HR sizes for the period ranged from 0.25 to 5.20 km2 (mean = 2.30 km2. Two thirds of the tagged G. morhua were infected with black spot disease Cryptocotyle lingua parasites; these fish had larger HRs and occupied deeper water compared with non-infected fish. The infected fish also tended to be more active in terms of horizontal swimming. From an ecological and evolutionary perspective, any environmental change that modifies G. morhua behaviour may therefore also alter the parasite load of the population, and its conservation and fishery status.

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

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.