Relative helminth size in crustacean hosts: in vivo determination, and effects of host gender and within-host competition in a copepod infected by a cestode

Crustaceans are important hosts for a number of helminth parasites, and they are increasingly used as models for studying the physiology, ecology and evolution of parasite-host interactions. In ecological studies, this interaction is commonly described only in terms of prevalence and number of larvae per infected host. However, the volume of helminth parasites can vary greatly, and this variation can potentially give important insights into the nature of a parasite-host relationship. It may influence and be influenced, for example, by within-host competition, host size, growth, and life history. Here we present a simple method that allows rapid approximation of the absolute and relative volumes of cestode larvae within copepod hosts of various developmental stages (nauplii, copepodites and adults). The measurements are taken in vivo without much disturbance of the animals, i.e. the technique allows study of growth and development of the parasites in relation to that of their hosts. The principles of this technique can be adopted to other helminth parasites and other crustacean hosts. Using this method in the copepod Macrocyclops albidus infected with the cestode Schistocephalus solidus, we found that the relative parasite size (= `parasite index') ranged from 0.5% to 6.5% of host size 14 days after infection. It was greater in male than in female hosts. With increasing number of parasites per host, the total parasite volume increased while the mean volume of the individual parasites decreased. The magnitude of the observed parasite indices, the large variation that was found within a sample of 46 infected adult copepods, and the observed correlates suggest that this new index can indeed be an important measure of parasite success and its pathogenecity.     

Diapausing fertilized adults

Summary A lake population of the cyclopoid copepod Cyclops strenuus Fischer had a 1-year life cycle including a diapause from June to January. Dormant stages consisted of fertilized females, not previously described as a diapause stage, and a small fraction of cop V. The longevity of the sperm in a diapausing female was at least 6–7 months. Pre-fertilization is assumed to be an adaptive mechanism to assure fertilization when male abundance is low. This life cycle pattern also gives a shorter time-lag between diapause break and offspring development, and also the low total adult population exerts a lower predation pressure on their offspring. The flexibility of life cycle patterns of copepods is emphasized.     

Diapause of Cyclops vicinus (Uljanin) in Lake Søbygård: Indication of a Risk-spreading Strategy

Cyclops vicinus is reported to enter summer diapause triggered by day length in order to survive food scarcity and fish predation. Development ceases and the fourth or fifth copepodid stages persist in the sediment for several weeks. In Lake Søbygård, however, a small eutrophic lake in Denmark, C. vicinus is found in the sediment as well as in the open water throughout the year. We performed laboratory experiments to elucidate the life cycle strategy of this population. In order to find the factors that induce diapause we tested the influence of food composition and light regime on the proportion of copepods entering diapause. Further we examined the diapause pattern of the offspring from diapausing and non-diapausing parents to see whether the co-occurrence of diapausing and non-diapausing copepods results from genetic differentiation within the population. In all experiments some of the copepods developed directly into adults, while others remained at the fourth or fifth copepodid stage and displayed diapause features. The proportion of copepods that developed directly into adults was influenced by food supply. Fewer individuals entered diapause when they received a mixed diet of algae and ciliates or algae, ciliates and seston compared with a pure algal diet. The response to light was different than described in literature: about 30% of the copepods entered diapause under dark conditions, and the diapause frequency was not higher when copepods had been exposed to continuous light. The offspring from diapausing parents showed no higher tendency to enter diapause than the offspring of non-diapausing parents, indicating no genetic differentiation. We conclude that diapause in C. vicinus represents a risk-spreading strategy, modified by food. The simultaneous production of diapausing and non-diapausing offspring ensures survival under harsh conditions; the influence of the food supply on the proportion of individuals entering diapause may adjust the population to the actual environmental condition. This flexible life cycle strategy might contribute to the exclusive dominance of C. vicinus in Lake Søbygård.     

Coexistence with similar life cycles in two species of freshwater copepods (Crustacea)

Two species of freshwater cyclopoid copepods, Cyclops scutifer Sars and Mesocyclops leuckarti (Claus), with different ecological requirements, coexist in three humic ponds near the timberline in boreal forests in south-central Norway. The life cycle in both species is basically regulated by complete depletion of oxygen during the long winter. There is a short period of reproduction in spring followed by a rapid development during summer and autumn to advanced instars, capable of diapause when oxygen depletion starts in early winter. Both species have an exclusive 1-yr cycle.The long winter under anoxic conditions is passed in diapause as large copepodids (late developmental instars) with usually no individuals left in the plankton.Even if co-occurring in the same habitats with similar life cycles, there is a temporal and spatial segregation.     

Competitive Growth Strategies in Intermediate Hosts: Experimental Tests of a Parasite life-History Model Using the Cestode, Schistocephalus solidus

In parasites with a complex life cycle, the fitness of an individual depends on its probability of reaching the final host and on its fecundity. Because larval growth in intermediate hosts may affect both transmission and adult size, selection should optimize growth patterns that are conditional on the presence and number of conspecific competitors. A recent model predicts that the total parasite volume per host should increase with intensity if larvae are able to vary growth depending on the number of conspecifics in the host (Life History Strategy hypothesis, i.e. LHS). Further, we would here expect growth rates to increase with intensity. By contrast, under the simplest alternative hypothesis of Resource Constraints (i.e. RC), the total parasite volume should remain constant. We experimentally infected copepods Macrocyclops albidus with the cestode Schistocephalus solidus to achieve 1, 2 or 3 parasites per host taking care that hosts had similar quality status at each infection level, and compared larval growth trajectories at the three intensity levels. The asymptotic total parasite volume was larger in double and triple infections than in single infections. Furthermore, the asymptotic total parasite volume was significantly larger in triple than in double infections but only in larger copepods that were less constrained by a host-size ceiling effect. These results, together with the fact that growth rates increased with intensity, support the LHS hypothesis: procercoids of a tapeworm may “count” their conspecific competitors in their first intermediate host to harvest its resources strategically until the next step in their complex life cycle. Co-ordinating editor: A. Biere     

Reef Fishes Have Higher Parasite Richness at Unfished Palmyra Atoll Compared to Fished Kiritimati Island

We compared parasite communities at two coral atolls in the Line Islands chain of the central Pacific (Kiritimati Island and Palmyra Atoll). Palmyra Atoll is relatively pristine while Kiritimati Island is heavily fished. At each island, we sampled five fish species for helminth and arthropod endoparasites: Chromis margaritifer, Plectroglyphidodon dickii, Paracirrhites arcatus, Acanthurus nigricans, and Lutjanus bohar. The surveys found monogeneans, digeneans, cestodes, nematodes, acanthocephalans, and copepods. Parasite richness was higher at Palmyra compared to Kiritimati for all five fish species. Fishes from Palmyra also tended to have more parasites species per host, higher parasite prevalence, and higher parasite abundance than did fishes from Kiritimati. The lower parasitism at Kiritimati may result from a simplified food web due to over fishing. Low biodiversity could impair parasite transmission by reducing the availability of hosts required by parasites with complex life cycles. Most notably, the lower abundances of larval shark tapeworms at Kiritimati presumably reflect the fact that fishing has greatly depleted sharks there in comparison to Palmyra.     

A simple method for maintaining a laboratory supply of limnetic cyclopoid copepods

The following method is used for maintaining a laboratory supply of freshwater cyclopoid copepods with intervening diapause phase. Animals are collected with a core sampler from the upper sediment surface just after initiation of diapause, i.e. during early summer for summer resting species and mid autumn for winter resting species. The method makes it possible at all times of the year to keep a variety of cyclopoid copepod species ready for experimental work. The following widespread species have been used: Mesocyclops leuckarti, Thermocyclops oithonoides, Diacyclops bicuspidatus, Cyclops vicinus, C. strenuus and C. abyssorum. Probably all cyclopoid copepods with a diapause phase could be stored according to the present method.     

The biology and morphology of the marine harpacticoid copepod Heteropsyllus nunni Coull, during encystment diapause

Heteropsyllus nunni Coull, a meiobenthic harpacticoid copepod is the marine crustacean to undergo a state of diapause within a cyst. A 12 month field study indicated H. nunni adults reached peak population densities in winter, with nauplii maturing in the spring, becoming adults by April or May.At the last stage of development, a mature but unmated adult, they begin to prepare for encystment diapause. The copepods remain within their cyst in a state of diapause for 3–4 months during the summer only. Studies on the effects of temperature and photoperiod suggested that these two environmental cues are not crucial for induction or termination of diapause. Low temperature delayed development and time to encystment, while high temperatures accelerated development, making the time to encystment shorter. There were males than females in the cysts in laboratory experiments. Upon excystment, the copepods mate, and females begin egg production within one week. Adults that have excysted and mated die after a few weeks of active reproductive effort. Nauplii go on to mature and begin the univoltine diapause/reproductive cycle.The copepods prepare for dormancy in two ways: they begin to produce and store two types of secretory products to be used in cyst construction; then they produce large quantities of lipid to be used as a nutrient supply throughout diapause. Histochemistry of the cyst-building material indicated the lower urosome is full of two chemically different products. Dorsally, there is a storage sac of proteinaceous material. The ventral sac of secretory product is a mucopolysaccharide. The copepod builds the spherical cysts in a matrix of small and large sand grains. The cysts fit tightly around the ventral portion of the animal in the its flexed position: however, there is a large space between the cyst and the sides of the copepod.Biochemical analysis of the cyst showed it is composed of an amino acid complex similar to collagenous material. Scanning electron microscopy revealed a complex of large cuticular pores located in the lower urosome and caudal rami. There are specific pores for secretion of the two cyst-building products.     

Effect of Ascaridia compar infection on rock partridge population dynamics: empirical and theoretical investigations

Helminth parasites have the potential to significantly affect the dynamics of their hosts. As a consequence, they can dramatically threaten the persistence of endangered species, such as rock partridge Alectoris graeca saxatilis, found in the Province of Trento (northern Italy). The aim of this work was to understand the effect of helminth parasites on rock partridge fitness, and the subsequent potential effects on host population dynamics. In particular, we investigated the hypothesis that infections from Ascaridia compar induce rock partridge population cycles observed in Trentino. In order to support this hypothesis, we compared the predictions obtained from a host–parasite interaction model including variable parasite aggregation with multi‐annual empirical data of A. compar infection in natural host populations. We estimated host demographic parameters using rock partridge census data from Trentino, and the parasitological parameters from a series of experimental infections in a captive rock partridge population. The host–parasite model predicted higher A. compar abundance in rock partridge populations exhibiting cyclic dynamics compared to non‐cyclic ones. In addition, for cyclic host populations, the model predicted an increase in mean parasite burden with the length of cycle period. Model predictions were well‐supported by field data: significant differences in parasite infection between cyclic and non‐cyclic populations and among cyclic populations with different oscillation periods were observed. On the basis of these results, we conclude that helminth parasites can not be ruled out as drivers of rock partridge population dynamics in Trentino and must be considered when planning conservation strategies of this threatened species.     

The life cycle of Cyclops vicinus in Lake Søbygård: new aspects derived from sediment analyses

Cyclops vicinus is the only copepod species in the pelagic zone of Lake Søbygård and can be found there all year round. We studied the population dynamics of this copepod over a one-year period. In contrast to earlier studies we included the copepods in the pelagic zone as well as the copepods resting in the sediment. Cyclops vicinuswas found not only in the open water, but also in the sediment throughout the year. From the fluctuations of the abundances in both habitats we suggest that the life cycle is more complex than known so far: different diapause pattern appear within the population and the induction of diapause is different than in other populations. We assume that these features contribute to the predominance of C. vicinusin Lake Søbygård.     

Genomic linkage map of the human blood fluke Schistosoma mansoni

Background  

Schistosoma mansoni is a blood fluke that infects approximately 90 million people. The complete life cycle of this parasite can be maintained in the laboratory, making this one of the few experimentally tractable human helminth infections, and a rich literature reveals heritable variation in important biomedical traits such as virulence, host-specificity, transmission and drug resistance. However, there is a current lack of tools needed to study S. mansoni's molecular, quantitative, and population genetics. Our goal was to construct a genetic linkage map for S. mansoni, and thus provide a new resource that will help stimulate research on this neglected pathogen.     

Genetic manipulation allows in vivo tracking of the life cycle of the son-killer symbiont,Arsenophonus nasoniae,and reveals patterns of host invasion,tropism and pathology

Maternally heritable symbionts are common in arthropods and represent important partners and antagonists. A major impediment to understanding the mechanistic basis of these symbioses has been lack of genetic manipulation tools, for instance, those enabling transgenic GFP expression systems for in vivo visualization. Here, we transform the ‘son-killer’ reproductive parasite Arsenophonus nasoniae that infects the parasitic wasp Nasonia vitripennis with the plasmid pOM1-gfp, re-introduce this strain to N. vitripennis and then used this system to track symbiont life history in vivo. These data revealed transfer of the symbiont into the fly pupa by N. vitripennis during oviposition and N. vitripennis larvae developing infection over time through feeding. A strong tropism of A. nasoniae to the N. vitripennis ovipositor developed during wasp pupation, which aids onward transmission. The symbiont was also visualized in diapause larvae. Occasional necrotic diapause larvae were observed which displayed intense systemic infection alongside widespread melanotic nodules indicative of an active but failed immune response. Our results provide the foundation for the study of this symbiosis through in vivo tracking of the fate of symbionts through host development, which is rarely achieved in heritable microbe/insect interactions.     

Extended life cycle in the chestnut weevil: prolonged or repeated diapause?

Many insect species extend the life cycle of a part of their population over several years. The adaptive value of these long cycles is now well documented, but the physiological processes underlying them have been little studied. Long life cycles are usually viewed as resulting from prolonged diapause proceeding from a simple extension of the usual winter diapause. However, this hypothesis has not been greatly tested, and information is lacking for species with a larval diapause. The energetic cost of a prolonged larval diapause also needs to be measured, because the few published estimates of lipid consumption concern an imaginal diapause. It is not therefore clear whether the negligible lipid consumption observed during adult prolonged diapause can be extrapolated to larval diapause. From microrespirometry and lipid measures in the chestnut weevil, Curculio elephas Gyllenhal (Coleoptera: Curculionidae), we show that: (1) in contrast to the usual hypothesis, the long cycle does not result from an extension of larval winter diapause, but is due to a prolonged diapause occurring secondarily to a developmental phase, and (2) energy consumption during the prolonged diapause is not negligible, but is provided for by a higher initial lipid content in the long cycle individuals. The adaptive value of the observed cycle is discussed.     

QTL analysis of the photoperiodic response and clinal distribution of period alleles in Nasonia vitripennis

In seasonal environments, organisms synchronize their life cycle with the annual cycle of environmental factors. In many insect species, this includes a diapause response: a timed dormant stage that allows to survive harsh winter conditions. Previously, we have shown that larval diapause in the parasitic wasp Nasonia vitripennis is induced by the mother upon exposure to a threshold number of short photoperiods (named switch point) and diapause response follows a latitudinal cline in natural populations. Here, we present a QTL analysis using two lines derived from the extremes of this clinal distribution: a northern line from Oulu, Finland and a southern line from Corsica, France. A genomic region on chromosome 1 and one on chromosome 5 were found to be associated with photoperiodic diapause induction. Interestingly, these regions contain the putative clock genes period, cycle (chromosome 1) and cryptochrome (chromosome 5). An analysis of period polymorphisms in seven European populations showed a clinal distribution of two main haplotypes that correlate with the latitudinal cline for diapause induction.     

Adaptions in the life cycle ofDermacentor variabilis (Say) andIxodes dammini (Spielman,Clifford, Piesman,and Corwin) marginal populations (Acari: Ixodidae)

Both theDermacentor variabilis andIxodes dammini adult diapause seasons are followed by a breeding period. At the edge of the ticks' range, where diapause and breeding occur under adverse climatic conditions, the species still conserves its fixed response for the onset of diapause. As a result, most of the following breeding effort is wasted; the population can only be maintained by an aberrant breeding period during the normal diapause season.When the diapause and breeding periods are suitable for the climate at the edge of the ticks' range, the rate of immature development versus the temperature regime can result in the appearance of the immature stages out of phase with the timing for the optimal life cycle in the marginal climate. The ticks are limited to minor adjustments of their development rates but, if this is all that is required for an optimum life cycle, this response can preadapt the species to the foreign climate.     

Control of Diapause by Acidic pH and Ammonium Accumulation in the Hemolymph of Antarctic Copepods

Life-cycles of polar herbivorous copepods are characterised by seasonal/ontogenetic vertical migrations and diapause to survive periods of food shortage during the long winter season. However, the triggers of vertical migration and diapause are still far from being understood. In this study, we test the hypothesis that acidic pH and the accumulation of ammonium (NH₄ ⁺) in the hemolymph contribute to the control of diapause in certain Antarctic copepod species. In a recent study, it was already hypothesized that the replacement of heavy ions by ammonium is necessary for diapausing copepods to achieve neutral buoyancy at overwintering depth. The current article extends the hypothesis of ammonium-aided buoyancy by highlighting recent findings of low pH values in the hemolymph of diapausing copepods with elevated ammonium concentrations. Since ammonia (NH₃) is toxic to most organisms, a low hemolymph pH is required to maintain ammonium in the less toxic ionized form (NH₄ ⁺). Recognizing that low pH values are a relevant factor reducing metabolic rate in other marine invertebrates, the low pH values found in overwintering copepods might not only be a precondition for ammonium accumulation, but in addition, it may insure metabolic depression throughout diapause.     

Parasite-induced Host Mortality: Indirect Evidence From a Long-term Study

A long-term field study of a perturbed host–helminth system provides indirect evidence that a long-lived swimbladder nematode, Cystidicola farionis, induces mortality of Arctic charr, Salvelinus alpinus. The prevalence and abundance of this parasite has changed little over the period from 1987 to 1999. The cumulative numbers of L₃-stage larvae steadily increased with increasing host age, indicating a continuous exposure to infection throughout the life of the target fish host. Indirect methods, which used data pooled over years and long-term cohort analyses, indicate that parasite-induced host mortality (PIHM) occurs in hosts older than 10 years. Furthermore, using a short-term cohort method adjusted for worm recruitment, we found indications of PIHM occurrence even in younger age groups. These patterns do not seem to be caused by high parasite mortality rates since dead worms are rarely observed inside the swimbladder. Age-related changes in infection rates or in resistance to infection seem to play only a minor role as there were only slight changes in the preference of charr for feeding on amphipods (which are intermediate hosts) and in the acquisition rate of L₃ larvae in older hosts. Mortality of the most heavily infected hosts is the most probable explanation for the observed patterns.     

Parasite transfer from crustacean to fish hosts in the Lübeck Bight,SW Baltic Sea

Four helminth parasites out of 19 species found in the Lübeck Bight, Baltic Sea, were chosen for investigations on the transfer from invertebrate to small-sized fish hosts: larvae of the tapewormsSchistocephalus sp. andBothriocephalus sp. (Cestoda) living in planktonic copepods as primary hosts;Podocotyle atomon (Digenea) andHysterothylacium sp. (Nematoda) were found in benthic crustaceans, especiallyGammarus spp. These hosts were the prey of 3 gobiid fishes,Gobiusculus flavescens (feeding mainly on plankton),Pomatoschistus minutus (preferring benthos), andP. pictus (feeding more on plankton than benthos). Because the fishes selected smaller sizes of crustaceans, they ingested all stages of the copepods but only the smaller-sized groups of gammarids which were often less infested by parasites. In order to evaluate the probability for a fish to be parasitized by a helminth, an infestation potential index (IP) was calculated.Podocotyle atomon andHysterothylacium sp. revealed an IP which was far lower in gobies than expected when the prevalences of the previous hosts were taken into consideration. The IP of tapeworm larvae was mainly influenced by the feeding pressure of the gobiid predators, which might change with developmental stage and season. It is concluded that parasite transfer to the next host decreases when sizes of prey and predator differ only moderately. This mechanism can reduce the numbers of parasites transferred to less suitable or wrong hosts.