Behavioural adaptations of argulid parasites (Crustacea: Branchiura) to major challenges in their life cycle.

Fish lice (Argulus spp.) are obligate ectoparasites, which contrary to most aquatic parasites, retain the ability to swim freely throughout the whole of their life. In fish farms, they can quickly increase in numbers and without effective control cause argulosis, which results in the reduced growth and survival of their fish hosts. The morphology of Argulus spp, including their sensory organs, is suitable for both parasitism and free-swimming. By spending a considerable amount of time away from their host, these parasites risk being excessively dispersed, which could endanger mating success. Here we present a review of recent studies on the behaviour of Argulus spp, especially the aggregative behaviour that mitigates the dilution of the parasite population. Aggregation of parasites, which is especially important during the period of reproduction, occurs on different scales and involves both the aggregation of the host and the aggregation of the parasites on the host. The main behavioural adaptations of Argulus spp, including searches for hosts and mates, host manipulation and host choice, are all focused on the fish. As these ectoparasites repeatedly change hosts and inflict skin damage, they can act as vectors for fish pathogens. The development of environmentally friendly measures for the control and prevention of argulosis needs to take into account the behaviour of the parasites.

Trichodinid (Ciliophora: Trichodinidae) Infections in Perch (Perca fluviatilis) experimentally exposed to pulp and paper mill effluents.

Wild-caught European perch (Perca fluviatilis) were exposed in the laboratory to untreated bleached pulp and paper mill effluent in two separate experiments. The first experiment was conducted at 7-8°C using effluent concentrations of 5 and 10%, and the second experiment was conducted at ambient river temperature of 4-20°C using an effluent concentration of 1%. Trichodinid ciliates were identified and enumerated at the end of the exposure using a mucus subsampling technique from gill and skin as well as a formalin immersion technique, which provided total counts on each fish. Four different trichodinid species were identified on the fish. Prevalence of infection, mean number, and mean density of Trichodina spp. decreased on fish exposed to effluents compared with controls. Prevalence of infection, mean number, and mean density of Trichodinella epizootica decreased on fish exposed to 5% and 10% effluents but increased on fish exposed to 1% effluents compared with controls. These results demonstrate that trichodinid ciliates vary in their susceptibility to at least certain types of contaminants and cautions against using trichodinids as environmental indicators without delineating species.

Virulent and nonvirulent Flavobacterium columnare colony morphologies: characterization of chondroitin AC lyase activity and adhesion to polystyrene.

AIMS: Colony morphology variants of fish pathogenic Flavobacterium columnare were studied to clarify the role of colony morphology change in the virulence of the bacterium. Typical rhizoid colony (Rz) variants are virulent and moderately adherent, nonrhizoid rough (R) colony variants are nonvirulent and highly adherent, and soft colony (S) variants are nonvirulent and poorly adherent. METHODS AND RESULTS: Chondroitin AC lyase activity, adhesion to polystyrene at different temperatures and after modification of bacterial surface, and lipopolysaccharide (LPS) profiles of the variants were studied. The chondroitinase activity was significantly higher in the virulent, rhizoid variants than in the rough variants of the same strain. Temperature significantly increased the adhesion of rhizoid variants up to 20°C. Modification of bacterial surface suggested that adhesion molecules contain both carbohydrates and proteins. LPS did not differ between the variants of the same strain. CONCLUSIONS: The results suggest that in Fl. columnare both rhizoid colony morphology and high chondroitinase activity are needed for virulence and that temperature may promote the adhesion of the virulent variants to surfaces at fish farms. SIGNIFICANCE AND IMPACT OF THE STUDY: New information is produced on the virulence mechanisms of Fl. columnare and the reasons behind the survival of the bacterium at fish farms.

Eye fluke-induced cataracts in natural fish populations: is there potential for host manipulation?

Manipulation of host phenotype (e.g. behaviour, appearance) is suggested to be a common strategy to enhance transmission in trophically transmitted parasites. However, in many systems, evidence of manipulation comes exclusively from laboratory studies and its occurrence in natural host populations is poorly understood. Here, we examined the potential for host manipulation by Diplostomum eye flukes indirectly by quantifying the physiological effects of parasites on fish. Earlier laboratory studies have shown that Diplostomum infection predisposes fish to predation by birds (definitive hosts of the parasites) by reducing fish vision through cataract formation. However, occurrence of cataracts and the subsequent potential for host manipulation in natural fish populations has remained poorly explored. We studied the occurrence of eye fluke-induced cataracts from 7 common fish species (Gymnocephalus cernuus, Rutilus rutilus, Leuciscus leuciscus, Alburnus alburnus, Osmerus eperlanus, Coregonus lavaretus and Gasterosteus aculeatus) from the Bothnian Bay in the Baltic Sea. We found that the parasite-induced cataracts were common in fish and they also reached high levels which are likely to predispose fish to predation. However, we observed such cataracts only in species with the highest parasite abundances, which suggests that only certain hosts may be strongly affected by the infection.

Parasite-induced aggression and impaired contest ability in a fish host.

BACKGROUND: Success of trophically transmitted parasites depends to a great extent on their ability to manipulate their intermediate hosts in a way that makes them easier prey for target hosts. Parasite-induced behavioural changes are the most spectacular and diverse examples of manipulation. Most of the studies have been focused on individual behaviour of hosts including fish. We suggest that agonistic interactions and territoriality in fish hosts may affect their vulnerability to predators and thus the transmission efficiency of trophically transmitted parasites. The parasite Diplostomum spathaceum (Trematoda) and juvenile rainbow trout, Oncorhynchus mykiss, were used to study whether infection can alter aggression rates and territorial behaviour of intermediate fish hosts. RESULTS: The changes in behaviour of rainbow trout, Oncorhynchus mykiss, infected with an eye fluke Diplostomum spathaceum (Trematoda), was monitored over the course of an experimental infection for 1.5 months. At the beginning of their development, not yet infective D. spathaceum metacercariae decreased the aggressiveness of rainbow trout. By the time that metacercariae were fully infective to their definitive hosts, the aggressiveness increased and exceeded that of control fish. Despite the increased aggressiveness, the experimentally infected fish lost contests for a territory (dark parts of the bottom) against the control fish. CONCLUSIONS: The results obtained indicate that the parasitized fish pay the cost of aggressiveness without the benefit of acquiring a territory that would provide them with better protection against predators. This behaviour should increase transmission of the parasite as expected by the parasite manipulation hypothesis.

Intensive fish farming and the evolution of pathogen virulence: the case of columnaris disease in Finland.

Ecological changes affect pathogen epidemiology and evolution and may trigger the emergence of novel diseases. Aquaculture radically alters the ecology of fish and their pathogens. Here we show an increase in the occurrence of the bacterial fish disease Flavobacterium columnare in salmon fingerlings at a fish farm in northern Finland over 23 years. We hypothesize that this emergence was owing to evolutionary changes in bacterial virulence. We base this argument on several observations. First, the emergence was associated with increased severity of symptoms. Second, F. columnare strains vary in virulence, with more lethal strains inducing more severe symptoms prior to death. Third, more virulent strains have greater infectivity, higher tissue-degrading capacity and higher growth rates. Fourth, pathogen strains co-occur, so that strains compete. Fifth, F. columnare can transmit efficiently from dead fish, and maintain infectivity in sterilized water for months, strongly reducing the fitness cost of host death likely experienced by the pathogen in nature. Moreover, this saprophytic infectiousness means that chemotherapy strongly select for strains that rapidly kill their hosts: dead fish remain infectious; treated fish do not. Finally, high stocking densities of homogeneous subsets of fish greatly enhance transmission opportunities. We suggest that fish farms provide an environment that promotes the circulation of more virulent strains of F. columnare. This effect is intensified by the recent increases in summer water temperature. More generally, we predict that intensive fish farming will lead to the evolution of more virulent pathogens.

Vertebrate diets derived from trophically transmitted fish parasites in the Bothnian Bay.

Parasites that are transmitted through predator-prey interactions may be used as indicators of trophic relationships between organisms. Yet, they are rarely used as such in the construction of topological (predator-prey) food webs. We constructed food webs of vertebrate trophic interactions using observed diet alone, trophically transmitted parasites alone, and the combination of the two based on data from 31 species of fish from the Bothnian Bay, Finland. The fish food web contained 530 links derived from observed diet, 724 links inferred from parasitism, and 1,058 links calculated from a combination of both stomach contents and parasites. This sub-web constructed from stomach contents had a mean of 17.1 links per fish species, while that using parasites had 23.4 links per fish. Combining the two diet indicators yielded 34.1 links per fish species, illustrating the complementarity of the two methods. Mean number of prey species per fish species was 12.5 using observed diet items, 15.8 using parasites, and 24.5 using both measures together. Mean number of predators per fish species was 7.4 using observed diet, 11.7 using parasites and 15.0 using both. A positive correlation was found between the mean number of parasites and the number of prey taxa in the diet among the fishes. Omnivorous fish had the highest diversity of both parasite species and prey items, while benthophagous fish had among the lowest. Mean total abundance and mean total prevalence of parasites correlated positively with fish size, with piscivores being the largest with the highest abundance and prevalence, while planktivores and benthivores had the lowest. Trophically transmitted parasites may be used to help construct vertebrate sub-webs and derive information about food web processes. Parasites alone provided equivalent if not more information than observed diet. However, resolution is improved by using parasites and observed diet together.

Being successful in the world of narrow opportunities: transmission patterns of the trematode Ichthyocotylurus pileatus.

Parasites with complex life cycles face 2 major challenges for transmission in northern latitudes. They have to cope with the general unpredictability associated with the series of transmission events required for completion of the cycle, and transmission has to be completed within a narrow temporal window because of strong seasonality. Despite this, some parasites show high transmission success, suggesting the operation of effective transmission mechanisms. We explored the transmission of Ichthyocotylurus pileatus (Trematoda) from its snail (Valvata macrostoma) to fish (Perca fluviatilis) hosts by examining some key characteristics in the dynamics of the cercarial emergence from snails. Transmission took place within a few weeks mainly in July, thus verifying the narrow temporal window for transmission. The output of the short-lived cercariae from the snails was low and variable in magnitude, but nevertheless resulted in a rapid and high rate of infection in newly hatched fish. The cercarial emergence showed a strong circadian rhythm with most of the cercariae emerging in early evening and night, which might represent the most likely mechanism underlying the high rate of transmission in this species. We emphasize the importance of holistic approaches combining aspects of multiple host species in studies on transmission of complex life-cycle parasites.

Acanthocephalan size and sex affect the modification of intermediate host colouration.

For trophically transmitted parasites, transitional larval size is often related to fitness. Larger parasites may have higher establishment success and/or adult fecundity, but prolonged growth in the intermediate host increases the risk of failed transmission via natural host mortality. We investigated the relationship between the larval size of an acanthocephalan (Acanthocephalus lucii) and a trait presumably related to transmission, i.e. altered colouration in the isopod intermediate host. In natural collections, big isopods harboured larger worms and had more modified (darker) abdominal colouration than small hosts. Small isopods infected with a male parasite tended to have darker abdominal pigmentation than those infected with a female, but this difference was absent in larger hosts. Female size increases rapidly with host size, so females may have more to gain than males by remaining in and growing mutually with small hosts. In experimental infections, a large total parasite volume was associated with darker host respiratory operculae, especially when it was distributed among fewer worms. Our results suggest that host pigment alteration increases with parasite size, albeit differently for male and female worms. This may be an adaptive strategy if, as parasites grow, the potential for additional growth decreases and the likelihood of host mortality increases.

Differences in parasite susceptibility and costs of resistance between naturally exposed and unexposed host populations.

It is generally assumed that resistance to parasitism entails costs. Consequently, hosts evolving in the absence of parasites are predicted to invest less in costly resistance mechanisms than hosts consistently exposed to parasites. This prediction has, however, rarely been tested in natural populations. We studied the susceptibility of three naïve, three parasitized and one recently isolated Asellus aquaticus isopod populations to an acanthocephalan parasite. We found that parasitized populations, with the exception of the isopod population sympatric with the parasite strain used, were less susceptible to the parasite than the naïve populations. Exposed but uninfected (resistant) isopods from naïve populations, but not from parasitized populations, exhibited greater mortality than controls, implying that resistance entails survival costs primarily for naïve isopods. These results suggest that parasites can drive the evolution of host resistance in the wild, and that co-existence with parasites may increase the cost-effectiveness of defence mechanisms.

Seasonal changes in host phenotype manipulation by an acanthocephalan: time to be transmitted?

SUMMARY: Many complex life cycle parasites exhibit seasonal transmission between hosts. Expression of parasite traits related to transmission, such as the manipulation of host phenotype, may peak in seasons when transmission is optimal. The acanthocephalan Acanthocephalus lucii is primarily transmitted to its fish definitive host in spring. We assessed whether the parasitic alteration of 2 traits (hiding behaviour and coloration) in the isopod intermediate host was more pronounced at this time of year. Refuge use by infected isopods was lower, relative to uninfected isopods, in spring than in summer or fall. Infected isopods had darker abdomens than uninfected isopods, but this difference did not vary between seasons. The level of host alteration was unaffected by exposing isopods to different light and temperature regimes. In a group of infected isopods kept at 4 degrees C, refuge use decreased from November to May, indicating that reduced hiding in spring develops during winter. Keeping isopods at 16 degrees C instead of 4 degrees C resulted in higher mortality but not accelerated changes in host behaviour. Our results suggest that changes in host and/or parasite age, not environmental conditions, underlie the seasonal alteration of host behaviour, but further work is necessary to determine if this is an adaptive parasite strategy to be transmitted in a particular season.

Spatial and temporal structure of the trematode component community in Valvata macrostoma (Gastropoda, Prosobranchia).

We conducted the first comprehensive study on the spatiotemporal structure of trematode communities in the large-mouthed valve snail, Valvata macrostoma. A total of 1103 snails were examined monthly between May and October 2007 from Lake Konnevesi, Central Finland, from a shallow (1-2 m deep) and an offshore site (5-6 m deep), located ca. 50-70 m apart. Snails were infected by 10 trematode species. The species composition and prevalence were strikingly different between the sites with high species diversity in the shallow site (all 10 species; total prevalence of sporocysts/rediae 12.1%, metacercariae 55.4%) compared to the deeper site (3 species; prevalence 15.0% and 1.9%, respectively). This difference persisted throughout our study and is probably related to the spatial distribution of bird definitive hosts, whereas the seasonal parasite dynamics are likely to be affected by changes in the age-structure of the snail population. The probability of sporocyst infections increased with snail size, but no such trend was observed in redial or metacercarial infections which decreased with host size. Our results show that generally well-described spatiotemporal differences in trematode infection of molluscs can emerge in very narrow spatial and temporal scales, which emphasizes the importance of these factors in community studies.

The effect of Echinorhynchus borealis (Acanthocephala) infection on the anti-predator behavior of a benthic amphipod.

In benthic habitats, predators can generally not be detected visually, so olfaction may be particularly important for inducing anti-predation behaviors in prey organisms. Manipulative parasites infecting benthic hosts could suppress these responses so as to increase the probability of predation and thus trophic transmission. We studied how infection with the acanthocephalan Echinorhynchus borealis affects the response of the benthic amphipod Pallasea quadrispinosa to water conditioned by burbot (Lota lota), the parasite's definitive host. In normal lake water, refuge use by infected and uninfected amphipods was similar, but when exposed to burbot-conditioned water, uninfected amphipods spent much more time hiding than infected amphipods. Thus, rather than affecting ambient hiding behavior, E. borealis infection seems to alter host response to a predator. A group of amphipods sampled from a postglacial spring that is devoid of fish predators exhibited only a weak response to burbot-conditioned water, perhaps suggesting these anti-predator behaviors are costly to maintain. The hiding behavior of spring and infected amphipods was very similar. If the reduced refuge use by the spring amphipods reflects adaptation to a predator-free environment, this indicates that E. borealis severely weakens its host's anti-predator behavior. Presumably this increases the likelihood of parasite transmission.

Multidimensionality and intra-individual variation in host manipulation by an acanthocephalan.

Trophically-transmitted parasites frequently alter multiple aspects of their host's phenotype. Correlations between modified characteristics may suggest how different traits are mechanistically related, but these potential relationships remain unexplored. We recorded 5 traits from individual isopods infected with an acanthocephalan (Acanthocephalus lucii): hiding, activity, substrate colour preference, body (pereon) coloration, and abdominal (pleon) coloration. Infected isopods hid less and had darker abdominal coloration than uninfected isopods. However, in 3 different experiments measuring hiding behaviour (time-scales of observation: 1 h, 8 h, 8 weeks), these two modified traits were not correlated, suggesting they may arise via independent mechanisms. For the shorter experiments (1 h and 8 h), confidence in this null correlation was undermined by low experimental repeatability, i.e. individuals did not behave similarly in repeated trials of the experiment. However, in the 8-week experiment, hiding behaviour was relatively consistent within individuals, so the null correlation at this scale indicates, less equivocally, that hiding and coloration are unrelated. Furthermore, the difference between the hiding behaviour of infected and uninfected isopods varied over 8 weeks, suggesting that the effect of A. lucii infection on host behaviour changes over time. We emphasize the importance of carefully designed protocols for investigating multidimensionality in host manipulation.

Monogenean assemblages and the apparent transmission capability of monogeneans between related fish species: an experimental study.

Gyrodactylid monogeneans are considered to be the most invasive fish parasites. They are omnipresent on teleost fishes and host change might play a crucial role in their biology. In the present study, related fish species were kept in single- or mixed-species groups to test the ability of Gyrodactylus species to change their host fish under experimental conditions. The first group included two percid fish species, perch Perca fluviatilis L. and ruffe Gymnocephalus cernuus (L.), and the second group two cyprinid species, roach Rutilus rutilus (L.) and minnow Phoxinus phoxinus (L.). A total of 4,182 specimens of 19 monogenean species belonging to five genera were observed, and eight species of Gyrodactylus were identified. There were three species of Gyrodactylus found on roach and six species on minnow. Gyrodactylus vimbi parasitized both cyprinid fish species and was the only species that increased in intensity during the experiments and also assumed to transmit from minnow to roach, while the other recorded host change case, Gyrodactylus macronychus, was represented by a single individual that transmitted from minnow to roach. Gyrodactylus rutilensis and Gyrodactylus pannonicus remained on their original hosts (roach and minnow, respectively). Gyrodactylus cernuae was the only species observed on both ruffe and perch, but ruffe appeared more susceptible to this parasite. Most of the gyrodactylids observed on roach and minnow were situated on the fins, while gyrodactylids of perch and ruffe parasitized mainly the gill chamber and head.

Host specificity of Argulus coregoni (Crustacea: Branchiura) increases at maturation.

We tested the hypothesis that host specificity in ectoparasites does not depend exclusively on the features of the host but also on surrounding habitats, using 2 fish ectoparasites, Argulus coregoni and A. foliaceus (Crustacea: Branchiura), occurring sympatrically in Finnish lakes. Although these parasites are considered to be of low specificity, we found that the larger of the 2 species, A. coregoni developed a pronounced preference for salmonid hosts at the beginning of maturation (defined by the presence of copulating specimens). Argulus foliaceus infects a much wider range of fish hosts. We showed that specialization of A. coregoni on salmonids does not necessarily result from incompatibility with other fishes, but could instead reflect higher sensitivity of oxygen depletion compared with A. foliaceus. Adult A. coregoni may meet these demands by attaching to salmonids, the typical inhabitants of well-aerated waters. Young parasites of both species showed little host specificity and attached mainly to fishes with higher body reflectivity. In host choice experiments, A. coregoni of 4-5 mm length preferred salmonids (rainbow trout) to cyprinids (roach) irrespective of the type of fish host, on which it had been previously grown in the laboratory. We suggest that such an innate ontogenetic shift in host preference maintains the major part of the parasite population on its principal host, ensuring successful reproduction within suitable habitats.

Experimental evidence for a hierarchy of mate- and host-induced cues in a fish ectoparasite, Argulus coregoni (Crustacea: Branchiura).

Argulus coregoni is an ectoparasite primarily infesting freshwater salmonids. Sexually reproducing parasites such as A. coregoni are confronted with a dilemma between finding a mate and the costs involved in doing so; if mating partners are unavailable on a host, by leaving to search for a mate on a new host, the parasite is exposed to risks such as predation and energy loss. The utilization of chemical cues could enhance the probability of finding a host and/or a suitable mating partner and thus decrease the level of costs associated with detachment from the host. In this study we constructed a Y-maze arena to determine if adult A. coregoni respond to mate- and host-related chemical cues. We also tested the directional response towards light, since it has been suggested that photic cues are the most important cues for juvenile A. coregoni locating a host. Our results showed that both sexes were attracted to light and fish odour. Free-swimming A. coregoni males responded to chemical cues produced by adult females but not vice versa. The hierarchy of these stimuli was analyzed by pitting the cues against one another in the Y-maze, showing that light was the most salient stimulus for both male and female parasites. Moreover, male parasites were more strongly attracted towards light and fish odour than female odour. In another experiment in a semi-natural environment, we examined whether the ability of A. coregoni males to detect female odour influences their host choice. Free-swimming males did not preferentially infest fish infected with female parasites over parasite-free fish. We suggest that a hierarchy of stimulus responses exists, whereby free-swimming parasites first respond to host-related signals and most dominantly to visual cues. However, cues connected to mate finding may become a priority for late adult stages and/or once the parasite has attached to the host.

Interspecific and intraspecific interactions in the monogenean communities of fish: a question of study scale?

Monogenean communities of fish have generally been considered non-interactive as negative interspecific interactions have rarely been reported. Most of the earlier studies on monogenean communities, however, have been conducted not only in systems with relatively low parasite abundances but, more importantly, at study scales where microhabitat-level interactions between the parasites are easily overlooked. We examined the communities of 3 abundant Dactylogyrus (Monogenea) species on the gills of crucian carp (Carassius carassius) by analysing the interactions at the scale of individual gill filaments, where interactions between the species, if any, should most likely take place. Contrary to our expectations, we did not find evidence for competitive exclusion between the species, which suggests that monogenean communities are non-interactive even in high parasite abundances. At the species level, individual parasites were highly aggregated within the filaments, essentially showing a strong tendency to occur at either end of a filament. This, together with the result of differences in the distribution of juvenile parasites within the filaments compared to adults, suggests that these parasites are able to actively seek out their conspecifics in small-scale microhabitats during maturation, which again could enhance their mate-finding.

Phenotypic variation in infectivity of Diplostomum spathaceum cercariae within a population.

The present study examined phenotypic variation in infectivity of Diplostomum spathaceum (Trematoda) cercariae within a natural population. Twelve infected Lymnaea stagnalis were collected from the field, and the infectivity of cercariae from individual snails was assessed under constant laboratory conditions. At a water temperature of 16.3 C, the mean infectivity of cercariae from the snails varied between 55.5% and 87.5%. Depending on the source of variation, this may have important ecological and evolutionary implications for both natural parasite populations and those occurring in aquaculture.