Parasitism, oddity and the mechanism of shoal choice

When choosing between shoals differing in Schistocephalus solidus infection status, uninfected test sticklebacks Gasterosteus aculeatus showed a preference for joining uninfected conspecifics when shoal sizes were equal, but reversed this preference when the relative size of the infected shoal was increased by a factor of 3. When given a choice between a shoal composed of size-matched minnows Phoxinus phoxinus and a shoal composed of the same number of all uninfected or all S. solidus -infected sticklebacks, test fish always preferred the sticklebacks, regardless of their infection status, over the minnow shoal. These observations suggest that species, parasite status and shoal size are all of importance when fish decide which shoal to join.     

The effect of Schistocephalus solidus infection on meal size of three‐spined stickleback

The effect of infection with the pseudophyllidean cestode Schistocephalus solidus on the meal size of individually housed three‐spined stickleback Gasterosteus aculeatus was quantified. Infected fish harboured plerocercoid loads that contributed from 1·1 to 33·9% of their total mass. Across this range of infection levels, the presence of S. solidus infection had no significant directional effect on standard length ( L _S) corrected meal size of host three‐spined sticklebacks. Amongst multiply infected fish there was a significant negative relationship between L _S‐corrected meal size and the proportion of host mass contributed by S. solidus parasites. This relationship, however, did not hold for singly‐infected fish. Furthermore, the data suggest that multiply‐infected fish that harbour a combined mass of parasites contributing < c . 15% to host body mass might exhibit meal sizes that exceed those of length‐matched uninfected fish. The results suggest that although heavy infections can significantly reduce the meal size of heavily infected three‐spined sticklebacks, in the early stages of multiple S. solidus infections host food intake may increase. The probable causes of these differential effects on meal size and their consequences for the host‐parasite system are discussed.     

Schistocephalus solidus infections increase gonadotropins and gonadotropin releasing hormone (GnRH3) mRNA levels in the three-spined stickleback, Gasterosteus aculeatus

Parasites often impair the reproduction of their hosts, one well known case being the cestode Schistocephalus solidus which is a common parasite in three-spined sticklebacks, Gasterosteus aculeatus. One of the possible ways that this could be exerted is by suppression on the brain-pituitary-gonadal (BPG) axis. In this study, mRNA levels of FSH-β and LH-β and of GnRH2 (cGnRH II) and GnRH3 (sGnRH) were measured via Q-PCR in infected and uninfected fish sampled from the field a few weeks before the onset of breeding. The pituitary mRNA levels of both FSH-β and LH-β were higher in infected males than in uninfected males. Also in females, FSH-β mRNA levels were higher in infected individuals than in others, whereas there was no significant difference found in LH-β expression. Brain mRNA levels of GnRH3 were higher in infected fish than in uninfected fish in both sexes, but no difference was found in GnRH2 mRNA levels. Thus, infection by S. solidus was able to alter the expressions not only of gonadotropins (GtHs), but also of GnRH which has not been observed previously. However, the effects are opposite to what should be expected if the parasite suppressed reproduction via actions on the brain-pituitary level. The gonads are perhaps more likely to be impaired by the parasites in other ways, and changed feedbacks on the BPG axis could then lead to the increases in GtHs and GnRH.     

Quantitative effects of three species of parasites on a population of Three-spined sticklebacks, Gasterosteus aculeatus

The effects of Schistocephalus solidus, Diplostomum gasterostei and Echinorhynchus clavula on a population of Three-spined sticklebacks from a pond in Somerset were studied. Schistocephalus was present at the highest level of infection and had the greatest effects on the sticklebacks. It caused the fish to weigh less than uninfected ones of the same length, as shown by the lower condition factor, and to grow more slowly; and it delayed the sexual maturation of fish of both sexes and prevented many from breeding. Diplostomum also had a significant effect on the condition of the sticklebacks and all three parasite species probably caused death or predation of heavily infected fish. The quantitative effects of the parasites on the condition, length and weight of the fish were calculated; seasonal changes in the condition factor could then be seen, and the growth rates for both length and weight calculated.     

Effects of Schistocephalus solidus infection on brain monoaminergic activity in female three-spined sticklebacks Gasterosteus aculeatus.

The three-spined stickleback Gasterosteus aculeatus is an intermediate host of the tapeworm Schistocephalus solidus. Changes in predator avoidance, foraging and shoaling behaviour have been reported in sticklebacks infested with S. solidus, but the mechanisms underlying parasite-induced behavioural changes are not understood. Monoamine neurotransmitters are involved in the control of behaviour and central monoaminergic systems are sensitive to various stressors. Thus, the behavioural effects of S. solidus infestation might be a reflection of changes in brain monoaminergic activity in the stickleback host. The concentrations of 5-hydroxytryptamine (5-HT), dopamine (DA), norepinephrine (NE) and their metabolites 5-hydroxy-indoleacetic acid (5-HIAA), homovanilic acid (HVA) and 3-methoxy-4-hydroxyphenylglycol (MHPG) were measured in the telencephalons, hypothalami and brainstems of parasitized and non-parasitized female sticklebacks held in the laboratory. The ratios of 5-HIAA:5-HT were significantly elevated in both the hypothalami and brainstems of infected sticklebacks. The concentrations of 5-HT and NE were significantly reduced in the telencephalons of infected fish as compared with controls, but there was no elevation of metabolite concentrations. The results are consistent with chronic stress in infected fish, but may also reflect other alterations of neuroendocrine status resulting from parasite infection.     

Nesting, courtship and kidney hypertrophy in Schistocephalus-infected male three-spined stickleback from an upland lake

The effect of Schistocephalus solidus infection on reproductive development and behaviour of male three-spined stickleback Gasterosteus aculeatus from an upland lake (Llyn Frongoch, Wales, U.K.) was investigated. Uninfected and infected males were collected on two separate occasions during the 2005 breeding season and encouraged to build nests under favourable laboratory conditions. Male nuptial colouration, courtship levels and nesting activity were recorded daily over two separate 21 day studies. Completed nests were removed, encouraging males to build multiple nests. On termination of each study, males were dissected and the kidney and any S. solidus plerocercoids were removed and weighed. In contrast to uninfected males, which readily built multiple nests, no infected males completed a nest during either study. Infected males also exhibited significantly reduced courtship levels, nesting activity, nuptial colouration, kidney development and body condition, compared with uninfected fish. Among infected three-spined stickleback, courtship and nesting behaviours tended to be more severely impacted in fish harbouring heavier plerocercoid burdens. In contrast to other populations, male three-spined stickleback from Llyn Frongoch appear to be unable to overcome the negative impacts of infections on reproductive development and behaviour when provided with short-term favourable conditions. Discrepancies between populations might have been related to the trophic status at the site of capture, emphasizing the importance of studying the biological effects of infections under different environmental conditions.     

Modulation of granulocyte responses in three-spined sticklebacks Gasterosteus aculeatus infected with the tapeworm Schistocephalus solidus

Leukocytes isolated from the head kidney and peripheral blood of 3-spined sticklebacks Gasterosteus aculeatus L. were analysed by means of flow cytometry during infection with the tapeworm Schistocephalus solidus (Müller, 1776). Although parasites increased their body weight continuously throughout the observation period (98 d), proportions of granulocytes increased in blood and head kidney only up to Day 63 post-infection (p.i.). Thereafter, declining proportions of granulocytes were observed in both organs. Thus the relative decrease in granulocyte number was not correlated to a decline in the parasitic load of the fish. To investigate a possible modulatory impact of S. solidus on granulocyte function, head kidney leukocytes were isolated at times before Day 63 p.i. and tested in vitro for their capacity to produce reactive oxygen species (ROS). Head kidney leukocytes from S. solidus-infected fish, analysed immediately after isolation (ex vivo, Day 40 p.i.), exhibited a higher ROS production when stimulated with phorbol myristate acetate (PMA), than leukocytes from naive, sham-treated control fish and fish that had resisted or cleared the infection (exposed but not infected). The latter showed an increased spontaneous ROS production that was not correlated to the numbers of granulocytes present in the head kidney isolates. In infected sticklebacks, spontaneous and PMA-induced ROS production was significantly correlated with numbers of granulocytes present in the head kidney isolates, suggesting that elevated ROS production was due to higher numbers of responding cells rather than an increased capacity of single cells. In vitro, after cultivation for 4 d in the presence of pokeweed mitogen (PWM) or extracts from S. solidus, head kidney leukocytes from control fish showed an increased ROS production and phagocytic activity compared with non-stimulated control cultures. In contrast, head kidney leukocytes from infected fish isolated on Days 48 and 44 p.i., failed to respond to S. solidus antigens in vitro. During S. solidus infection, granulocyte mobilisation resulted in elevated numbers of these cells in head kidneys, but the lack of an in vitro response to S. solidus antigens indicates an in vivo priming of granulocytes by the parasite. These observations may reflect the ability of S. solidus to impair the host's immune response once the parasite is developing in the body cavity of G. aculeatus.     

Effects of Schistocephalus solidus (Cestoda) on the food intake and diet of the three-spined stickleback, Gasterosteus aculeatus

Seasonal differences were detected between uninfected sticklebacks and those infected with Schistocephalus solidus in their stomach fullness and diet composition.     

A non-invasive morphometric technique for estimating cestode plerocercoid burden in small freshwater fish

A non-invasive morphometric technique is presented which can be used to predict the infection status and the proportion of infected fish weight contributed by parasite tissue in three-spined sticklebacks Gasterosteus aculeatus infected with plerocercoids of Schistocephalus solidus (Cestoda: Pseudophyllidea)     

The effect of hunger and cestode parasitism on the shoaling decisions of small freshwater fish

Individual minnows Phoxinus phoxinus and three-spined sticklebacks Gasterosteus aculeatus were provided with a mutually exclusive choice between joining a shoal of conspecifics and foraging alone in a maze. The shoaling decisions and foraging behaviour of individual fish were studied when the fish were satiated and after 24- and 48-h periods of food deprivation. Hunger level was found to have a significant effect on shoaling behaviour. When satiated, fish of both species spent a greater proportion of time within one body length of the shoal and spent less time out of visual contact with the shoal than after periods of food deprivation. The effect of the cestode parasite Schistocephalus solidus on the shoaling behaviour of stickleback hosts was complex. When satiated, infected fish spent less time than uninfected fish within one body length of the shoal, preferring to remain outside the shoal, yet within visual contact, although when food deprived there was no difference in the proportion of time spent by infected and uninfected fish close to the shoal. The possible ecological significance of this change in behaviour is discussed with reference to the manipulation hypothesis of host-parasite interactions.     

Annual variation in energy reserves in male three-spined stickleback, Gasterosteus aculeatm L. (Pisces, Gasterosteidae)

Wet and dry weights of tissue were measured and concentrations of glycogen, lipid and protein were estimated for the liver, gonad and carcass of male sticklebacks from an annual population collected each month over one complete year. In young-of-the-year there is one period of rapid weight gain, in all three body regions (liver, carcass and gonads) but particularly of the carcass, in the autumn and a second in spring and early summer. This is accompanied by an increase in the water content of all three body regions. The gonadosomatic index also increases sharply in spring and early summer. Young sticklebacks accumulate lipid and glycogen slowly during the autumn and winter of their first year of life and more rapidly from late winter to early summer. Thus, the period of most rapid accumulation of these reserves coincides with the time when body weight and gonad maturation are also increasing sharply. Lipid and glycogen levels fall during the reproductive season in those males that breed, so that by July they are reduced to 43% and 37% (respectively) of their peak values in May. Levels of protein increase throughout the year as the fish grow, but in breeding males by July the concentration of protein in the carcass falls to 70% of pre-breeding levels. Breeding males therefore reach the end of the reproductive season with their total energy reserves severely reduced, and consequently they suffer a very high mortality. In contrast, adult males that fail to reproduce survive beyond the breeding season. They continue to gain weight and to accumulate lipid and glycogen from August to September, but these energy reserves fall (to levels comparable to those of post-breeding fish) in December, when these fish probably die. These results demonstrate that in male sticklebacks, growth and gonad maturation can be sustained in parallel with the accumulation of energy reserves, which are subsequently extensively depleted as a result of reproductive activities.     

Parasite-associated growth enhancement in a fish-cestode system

Parasites impose an energetic cost upon their hosts, yet, paradoxically instances have been reported in which infection is associated with enhanced, rather than diminished, host growth rates. Field studies of these parasite effects are problematic, since the pre-infection condition of the hosts is generally unknown. Here, we describe a laboratory experiment in which the growth rate and body condition of 76 laboratory-reared three-spined stickleback fishes were examined before, during and after each fish was fed the infective stage of the parasitic cestode Schistocephalus solidus. Twenty-one of these fishes went on to become infected by the cestode. Fishes were individually housed and provided with an abundant food supply to eliminate the potentially masking effects of variable competitive ability. Infection occurred independently of fish gender, size, body condition or pre-exposure growth rate. After exposure to the cestode, infected fishes grew faster (excluding parasite weight) and maintained a similar or better body condition compared with uninfected fishes, despite developing enlarged spleens. The accelerated growth could not be explained by reduced gonadal development. This result, one of few demonstrations of parasite-associated growth enhancement in fishes, is discussed with respect to other such parasite systems.     

Variation in the reproductive potential of Schistocephalus infected male sticklebacks is associated with 11-ketotestosterone titre

Parasites can impact host reproduction by interfering with host endocrine systems, but the adaptive nature of such effects is disputed. Schistocephalus solidus plerocercoids are parasites of three-spined sticklebacks Gasterosteus aculeatus that are often associated with impaired host reproduction. Here, we relate reproductive behavior and physiology to levels of the androgen 11-ketotestosterone (11KT) in naturally infected and non-infected male sticklebacks from two UK populations. In one population infected males harbored heavy infections and showed uniformly reduced 11KT titres and kidney spiggin (nesting glue protein) content compared to non-infected fish. However in a second population infection levels were more variable and males with smaller infections recorded 11KT and spiggin titres that overlapped those of non-infected fish; among infected males from this population 11KT and kidney spiggin also both correlated negatively with infection severity. Male reproductive behavior correlated closely with 11KT titre in both populations, and infected males with high 11KT levels exhibited normal reproductive behavior. Our results suggest that Schistocephalus infection per se does not block reproductive development in male sticklebacks, and that some male fish may have the ability to breed whilst infected. Our results are not consistent with the hypothesis that Schistocephalus adaptively castrates male hosts via endocrine disruption; rather they support the hypothesis that reproductive disruption is a side effect of the energetic costs of infection.     

Host behaviour alteration by its parasite: from brain gene expression to functional test

Many parasites with complex life cycles modify their intermediate hosts'' behaviour, presumably to increase transmission to their final host. The threespine stickleback (Gasterosteus aculeatus) is an intermediate host in the cestode Schistocephalus solidus life cycle, which ends in an avian host, and shows increased risky behaviours when infected. We studied brain gene expression profiles of sticklebacks infected with S. solidus to determine the proximal causes of these behavioural alterations. We show that infected fish have altered expression levels in genes involved in the inositol pathway. We thus tested the functional implication of this pathway and successfully rescued normal behaviours in infected sticklebacks using lithium exposure. We also show that exposed but uninfected fish have a distinct gene expression profile from both infected fish and control individuals, allowing us to separate gene activity related to parasite exposure from consequences of a successful infection. Finally, we find that selective serotonin reuptake inhibitor-treated sticklebacks and infected fish do not have similarly altered gene expression, despite their comparable behaviours, suggesting that the serotonin pathway is probably not the main driver of phenotypic changes in infected sticklebacks. Taken together, our results allow us to predict that if S. solidus directly manipulates its host, it could target the inositol pathway.     

Sticklebacks from Greenland

Greenland freshwater sticklebacks Gasterosteus aculeatus were low-plated. Length distribution of 1 + and 2+ fish peaked at 33 and 46 mm, in early July. Pre-reproductive fish including 1 + fish were infected with Schistocephalus solidus . The diet consisted of chironomid larvae and pupae. Infected individuals ate smaller prey than non-infected ones, on one occasion where competition for food was likely to occur. Infected fish had lower stomach fullness, and the parasite compressed the stomach in heavily infected fish. The other fish species was charr, and the stickleback seemed to occupy a wide range of habitats.     

Effects of two parasites, Schistocephalus solidus (Cestoda) and Bunodera spp. (Trematoda), on the escape fast-start performance of three-spined sticklebacks

The fast‐start performance of three‐spined sticklebacks Gasterosteus aculeatus infected with Schistocephalus solidus and Bunodera spp. was determined and two distinct fast‐start responses (A and B) were observed. ‘A’ starts were of higher flexibility than B and three‐way ANOVA showed significant effects of A and B starts (P < 0·05), time (P < 0·05) and per cent standard body length, L_S (P < 0·05) on the orientation angle (angle of an individual segment of the fish with respect to the direction of travel). Schistocephalus solidus infection reduced maximum velocity (P < 0·05) and maximum acceleration (P < 0·05) of infected fish. Uninfected fish and fish infected with S. solidus up to a parasite index (parasite mass divided by the sum of fish and parasite mass) of 0·1 executed both types. Infected fish exclusively executed B starts for parasite index between 0·1 and 0·2. This was not due to a reduction in body flexibility associated with mechanical obstruction caused by S. solidus as no significant difference in the ratios of body width (P > 0·05) or depth (P > 0·05) to L_S were found between uninfected and infected fish. At a parasite index >0·2, infected fish were unable to perform escape fast‐starts increasing the likelihood of predation by their definitive hosts such as loons or belted kingfishers. Three‐spined sticklebacks infected with S. solidus with a parasite index of c. 0·2–0·3, however, were compromised by a suite of behavioural (e.g. increased foraging activity and amount of food consumed, increased risk associated with feeding and increased response latency to predatory stimuli), physiological (e.g. increased rate of oxygen consumption, slower growth, delayed sexual maturation and breeding success) and biomechanical (e.g. decreased fast‐start performance) factors. Bunodera spp. did not affect the escape fast‐start performance of three‐spined sticklebacks and no significant difference for maximum velocity (P > 0·05) and maximum acceleration (P > 0·05) was found.     

The clearance of hidden cestode infection triggered by an independent activation of host defense in a teleost fish

Parasites often elude effective recognition or attack (or both) by the host immune system, for example, though a tegument that possesses nonimmunogenic features. However, a general activation of host defense due to independent stimuli may increase immune activity to a level where such disguises are no longer effective, resulting in the clearance of an infection. We experimentally infected three-spined sticklebacks (Gasterosteus aculeatus) with the cestode Schistocephalus solidus. To independently foster a general immune response a few days later, we cut the tips of spines in some fish and sham-treated other fish. Cutting spines significantly reduced the prevalence of the infection. The injury evoked a physiological reaction that helped to clear a hidden parasite infection.     

The effect of parasitism on the toxicity of cadmium to the three-spined stickleback, Gasterosteus aculeatus L.

The toxicity of cadmium to sticklebacks was determined over a wide range of concentrations. The 96 h LC50 was found to be 6.5 mg Cd I⁻¹. The shape of the time-concentration curve suggests that cadmium may have two toxic mechanisms. The median period of survival for fish infected with the plerocercoids of the cestode, Schistocephalus solidus , was found to be considerably shorter than for non-parasitized fish. This observation is considered in the light of the known effects of 5. solidus on its host.     

Variation in stickleback head morphology associated with parasite infection

Parasites can affect host phenotypes, influencing their ecology and evolution. Host morphological changes occurring post-infection might result from pathological by-products of infection, or represent adaptations of hosts or parasites. We investigated the morphology of three-spined sticklebacks, Gasterosteus aculeatus , from a population naturally infected with Schistocephalus solidus , which grows to large sizes in their body cavity. We examined local effects of infection on trunk shape, which are imposed directly by the bulk of the growing parasite, and distant effects on head morphology. We show that trunk shape differed between infection classes, and was affected more severely in fish with heavier total parasite mass. We further show unexpected differences in head morphology. The heads of infected fish were reduced in size and differently shaped to those of non-infected fish, with infected fish having deeper heads. Importantly, both head size and shape were also affected more severely in fish with heavier total parasite mass. This latter result suggests that differences in morphology are caused by post-infection changes. Such changes may be incidental, evolutionarily neutral 'side effects' of infection. However, because head morphology affects foraging ecology, such changes are likely to have fitness consequences for hosts, and may constitute adaptations, either of hosts or of parasites. We discuss our finding in the context of the evolution of phenotypic plasticity, and suggest testable hypotheses examining the proximate mechanisms underlying these morphological effects and their potential evolutionary basis.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 759–768.     

Effect of concurrent infections of Lernaeocera Branchialis (Copepoda) and Trypanosoma murmanensis (Protozoa) on Atlantic cod, Gadus morhua

A study was initiated to ascertain the prevalence, effect and interaction of the adult stages of the parasitic copepod, Lernaeocera branchialis, on Atlantic cod concurrently infected with a hematozoan, Trypanosoma murmanensis, by comparing condition (K) factor, organ somatic indices, hematological values and lipid concentrations of the liver from infected and uninfected fish of comparable length. Prevalence of the copepod varied from 9 to 21% and was highest in fish examined on the southern coast of Newfoundland. Body condition and blood values were significantly lower in young cod infected with two parasites whereas only K-factor was altered in two size classes parasitized by one copepod. Although lipid concentrations were similar in three length groups of fish without and infected with L. branchialis, the pooled values were significantly greater among the infected group. Mortality occurred only in juvenile cod infected with the copepod and T. murmanensis concurrently. Cod which survived dual infections had lower K-factor and blood values than controls or fish harboring single, parasitic infections. It is concluded that an infection with an adult L. branchialis does not impair the health of fish in this area of the northwestern Atlantic Ocean, but the presence of T. murmanensis concurrently can potentiate its effect.