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Sites of female reproductive vulnerability: implications for testing and risk assessment.

All reproductive toxicity testing, whether in vivo or in vitro, should be conducted with consideration of the ultimate use of the data for reproductive risk assessment and the protection of human reproductive health. In this review selected sites of vulnerability in the female reproductive system are identified. These sites of female reproductive vulnerability may be utilized for in vitro toxicity testing assays, and the data from assays may be employed in the hazard identification and hazard characterization steps necessary for reproductive risk assessment. Using biomarkers of female reproductive function derived from in vitro toxicity testing it is possible to define functions that characterize female fecundity. These characterizations of female fecundity may be used to quantitate reproductive risk in human populations.

Animals↗

Metal-induced stress in bivalves living along a gradient of Cd contamination: relating sub-cellular metal distribution to population-level responses.

The use of biomarkers to assess the impacts of contaminants on aquatic ecosystems has noticeably increased over the past few years. Few of these studies, however, have contributed to the prediction of ecologically significant effects (i.e., at the population or community levels). The present field study was designed to evaluate the potential of metallothionein (MT) and sub-cellular metal partitioning measurements for predicting toxic effects at higher levels of the biological organization in freshwater bivalves (Pyganodon grandis) chronically exposed to Cd. For that purpose, we quantitatively sampled P. grandis populations in the littoral zone of nine lakes on the Precambrian Canadian Shield during two consecutive summers (1998 and 1999); lakes were characterized by contrasting Cd levels but similar trophic status. We tested relationships between the population status of P. grandis (i.e., growth parameters, density, biomass, secondary production, turnover ratio and cumulative fecundity) and (i) ambient Cd concentrations, (ii) sub-organismal responses (MT concentrations in the gill cytosol of individuals and Cd concentrations in three metal-ligand pools identified as M-HMW, the high molecular weight pool, M-MT, the metallothionein-like pool and M-LMW, the low molecular weight pool) and (iii) ecological confounding factors (food resources, presence of host fishes for the obligatory parasitic larval stage of P. grandis). Our results show that littoral density, live weight, dry viscera biomass, production and cumulative fecundity decreased with increasing concentrations of the free-cadmium ion in the environment (Pearson's r ranging from -0.63 to -0.78). On the other hand, theoretical maximum shell lengths (L( infinity )) in our populations were related to both the dissolved Ca concentration and food quality (sestonic C and N concentrations). Overall, Cd concentrations in the gill cytosolic HMW pool of the individual molluscs were the biomarker response that was most frequently and most strongly correlated with the population variables (Pearson's r ranging from -0.58 to -0.80). Our findings demonstrate, however, the difficulty of currently assigning to sub-cellular metal partitioning measurements (mainly Cd bound to the HMW fraction) any predictive role for population health, notably because of the influence of ecological confounding variables (e.g., the cumulative number of degree-days in the littoral zone, as is the case here). Metal contamination of our lakes has decreased markedly in the past 10 years and consequently we believe that the toxic effects of metals may have been replaced by some natural factors as the main agent for structuring the clam populations in these lakes.

Animals↗

Physiological cost of tolerance to toxicants in the European flounder Platichthys flesus, along the French Atlantic Coast.

Physiological and genetic responses of flounder Platichthys flesus populations were investigated along the French Atlantic Coast in one moderately contaminated estuary (Ster) and three contaminated estuaries (Seine, Loire and Gironde). The focus of this study was to explore the relationship between stress resistance and energetic trade-offs, in order to detect possible differential physiological capacities or performances between individuals carrying particular alleles or genotypes (allozyme data) characterised as "tolerant" or "sensitive". A general reduction of the relative fecundity, the growth rate and the condition factor was highlighted in contaminated fish populations, suggesting that survival in such polluted systems implies energetic costs for fish thus reducing the energy available for particular functions. A lower observed heterozygosity was also detected in contaminated populations with respect to the Ster, suggesting a general decrease in genetic variability in response to chemical stress (with an exception for the Seine estuary). This study confirmed the previously detected relationship between PGM 85, AAT1 95 alleles and reduced DNA damage in contaminated fish [Marchand, J., Tanguy, A., Laroche, J., Quiniou, L., Moraga, D., 2003. Responses of European flounder Platichthys flesus populations to contamination in different estuaries along the Atlantic coast of France. Mar. Ecol. Prog. Ser. 260, 273-284] and furthermore suggested that, reduced fecundity and condition factor associated to the individuals carrying the previous alleles, were also reflecting the cost of resistance to stress in polluted populations. The cost of tolerance to stress as well as the high gene flow from neighbouring populations less exposed to contamination may explain the apparently moderate increase of the suspected "tolerant" alleles in contaminated flounder populations.

Analysis of Variance↗

Effects of chronic internal alpha irradiation on physiology, growth and reproductive success of Daphnia magna.

Daphnids were chronically exposed to waterborne Am-241, an alpha-emitting radionuclide, ranging in concentration from 0.4 to 40 Bq ml(-1). Am-241 amounts were monitored in the medium, daphnid tissues and cuticles. Corresponding average dose rates of 0.02, 0.11 and 0.99 mGy h(-1) were calculated for whole organisms with internal alpha-radiation contributing 99% of total dose rates. Effects of internal alpha irradiation on respiration and ingestion rates, adult, egg and neonate individual dry masses, fecundity and larval resistance to starvation were examined in 23-day experiments. Daphnids showed increased respiratory demand after 23 days at the highest dose rate, suggesting increased metabolic cost of maintenance due to coping with alpha radiological stress. Although no effect was detected on ingestion rates between contaminated and control daphnids, exposure to dose rates of 0.11 mGy h(-1) or higher, resulted in a significant 15% reduction in body mass. Fecundity remained unchanged over the 23-day period, but individual masses of eggs and neonates were significantly smaller compared to the control. This suggested that increased metabolic expenditure in chronically alpha-radiated daphnids came at the expense of their energy investment per offspring. As a consequence, neonates showed significantly reduced resistance to starvation at every dose rate compared to the control. Our observations are discussed in comparison with literature results reported for cadmium, a chemical toxicant which affects feeding activity and strongly reduces individual energy uptake.

Age Factors↗

Inhibition of ovarian development by methyl farnesoate in the tadpole shrimp, Triops longicaudatus.

Methyl farnesoate (MF), a putative crustacean hormone, is the immediate precursor of insect juvenile hormone III (JHIII) in the biosynthetic pathway. We examined whether MF, shown to inhibit adult metamorphosis in several crustacean species, is a juvenilizing factor in the tadpole shrimp, Triops longicaudatus. Oocyte production was chosen as a parameter for measuring reproductive development. MF was administered to juveniles by ingestion via biological vector (Artemia nauplii), MF-coated food pellets, and MF liposome food pellets. Artemia were incubated in 30 microl of 5 microg/ml MF. The MF-coated and MF liposome pellets were prepared with MF concentrations ranging between 0.1 microg/g and 10 microg/g MF by weight. Groups of tadpole shrimp were treated with these vectors from the time of hatching for 5 or 10 days in laboratory and field studies. The treatment groups of all the MF vectors showed reductions in oocyte production. Lower concentrations of MF (0.75 microg/g-3.8 microg/g MF) appeared to have a physiological effect on fecundity, but higher concentrations (10 microg/g MF) reduced somatic growth. MF-coated pellets (1 microg/g MF) administered to adults (after 5 days) caused no difference in oocyte production. The observed reductions of fecundity and the disparity of results between MF treatment on juveniles and adults suggest that MF may regulate ovarian development.

Animals↗

Life-stage-dependent sensitivity of zebrafish (Danio rerio) to estrogen exposure.

The aim of this study was to identify periods in zebrafish (Danio rerio) development when estrogen exposure has long-term consequences on reproductive capabilities at the adult stage. To this end, zebrafish were exposed to 10 ng/L ethynylestradiol (EE(2)) during three stages of gonadal differentiation: (i) the juvenile hermaphroditic stage when gonads display the morphology of an immature ovary (in our zebrafish colony this lasted from 15 to 42 days post-fertilization [dpf]), (ii) the gonad transition stage when the hermaphroditic gonad differentiates into either testes or ovary (from 43 to day 71 dpf), and (iii) the premature stage of testicular and ovarian development (from 72 to 99 dpf). The consequences of stage-specific exposure to EE(2) were assessed by determining time to first spawning, fecundity (number of eggs per female per day), fertilization success (percentage of fertilized eggs) and sex ratio of the adults. Exposure during the gonad transition period induced a delay in the onset of spawning and a significant reduction of fecundity and fertilization success, whereas exposure during the hermaphroditic stage or during the premature stage had no significant impact on the reproductive parameters of adult fish. The results from this experiment pointed to the gonad transition stage as being most susceptible to persistent effects of developmental estrogen exposure. In a second experiment, the concentration dependency of the EE(2)effects was evaluated by exposing zebrafish during the gonad transition stage (43-71 dpf) to 1.67, 3 or 10 ng EE(2)/L. Significant effects of EE(2) on adult reproduction were found with 3 and 10 ng EE(2)/L, but not with 1.67 ng/L. Histological examination of the gonads revealed that at termination of EE(2) exposure (71 dpf), all individuals in the 3 and 10 ng EE(2)/L treatment possessed ovaries. However, this feminising effect appeared to be reversible since at the adult stage (190 dpf), both fish with ovaries and with testes were found. Thus, EE(2) exposure during the gonad transition stage seems to have no persistent effect on gonad histology but on reproductive capabilities.

Animals↗

Effects of pharmaceuticals on Daphnia survival, growth, and reproduction.

Pharmaceuticals have been globally detected in surface waters, and the ecological impacts of these biologically-active, ubiquitous chemicals are largely unknown. To evaluate the aquatic toxicity of individual pharmaceuticals and mixtures, we performed single species laboratory toxicity tests with Daphnia magna, a common freshwater zooplankton. We conducted acute (6-day) and chronic (30-day) exposure pharmaceutical bioassays and evaluated survivorship and morphology of adults and neonates, adult length, resting egg production, brood size (fecundity), and the proportion of male broods produced (sex ratio). In general, exposure to a single pharmaceutical in the 1-100 microg/l range yielded no apparent effects on the normal life processes of Daphnia. However, chronic fluoxetine exposure (36 microg/l) significantly increased Daphnia fecundity, and acute clofibric acid exposure (10 microg/l) significantly increased sex ratio. A mixture of fluoxetine (36 microg/l) and clofibric acid (100 microg/l) caused significant mortality; the same fluoxetine concentration mixed with 10 microg/l clofibric acid resulted in significant deformities, including malformed carapaces and swimming setae. Mixtures of three to five antibiotics (total antibiotic concentration 30-500 microg/l) elicited changes in Daphnia sex ratio. We conclude: (1) individual and mixtures of pharmaceuticals affect normal development and reproduction of Daphnia magna, (2) aquatic toxicity of pharmaceutical mixtures can be unpredictable and complex compared to individual pharmaceutical effects, and (3) timing and duration of pharmaceutical exposure influence aquatic toxicity.

Animals↗

Reproductive effects and bioconcentration of 4-nonylphenol in medaka fish (Oryzias latipes).

The objective of this study was to investigate the reproductive success (fertility and fecundity) of medaka (Oryzias latipes) exposed to 4-nonylphenol (4-NP) in water and the viability of their F(1) offspring. In addition, we measured the bioconcentration of 4-NP in eggs. After a 21-d exposure to 100microg/l 4-NP, medaka showed reduced egg production and fertility. Hepatic vitellogenin levels were increased significantly in males treated with 10, 50 and 100microg/l of 4-NP. In the F(1) generations, the hatchability and time to hatching of embryos in the 100microg/l treatment group were adversely affected, and 2-7microg 4-NP/g egg was found in spawned eggs (the bioconcentration factor: 30-100). These results indicate that (1) 4-NP adversely affected the reproduction (fecundity and fertility) of adult medaka, (2) 4-NP accumulated in eggs through maternal transfer and (3) these levels of 4-NP were associated with adverse effects in the F(1) offspring.

Animals↗

Chronic toxicity of tributyltin to development and reproduction of the European freshwater snail Lymnaea stagnalis (L.).

Chronic toxicity, growth and reproduction were measured in the freshwater gastropod Lymnaea stagnalis exposed to waterborne bis(tri-n-butyltin) oxide (TBTO) over a range of four nominal concentrations (0-10microg TBTl(-1)). Egg development was completely inhibited at 10microg TBTl(-1), whilst abnormal embryonic development was observed at 1microg TBTl(-1). For the solvent control and the 0.01microg TBTl(-1)treatment group, normal development of L. stagnalis was observed. Survivorship of hatchlings was significantly reduced by TBT at 1microgl(-1) while inhibition of shell growth of L. stagnalis was also observed at this concentration. The data were used to determine intrinsic growth rates (r) using two theoretical approaches (the Euler-Lotka equation and a Leslie Matrix). Both approaches showed that survival, fecundity and population growth rate were reduced at 1microg TBTl(-1). Interestingly, at 0.01microg TBTl(-1) snails showed a higher fecundity and growth rate than in the solvent control. The TBT concentration at which the r would equal zero (ECr(0)) and the population NOEC (No Observed Effect Concentration) were estimated. The population NOEC was defined as either the lower 95% confidence or lower 95% pointwise percentile limit of the ECr(0). Values obtained using the two different approaches were similar and thus a geometric mean was calculated to obtain a final representative population NOEC value for L. stagnalis of 2745ng TBTl(-1). The present data together with chronic toxicity TBT data for freshwater organisms, obtained from peer-reviewed literature, were used to construct a species sensitivity distribution (SSD). A predicted no effect concentration was then derived from the SSD (hazardous concentration at 5%, i.e., HC5 or 95% protection level). This SSD was compared with the SSD derived from saltwater species datasets. The HC5 value for saltwater species (3.55ng TBTl(-1); lower confidence limit: 1.93ng TBTl(-1)) was significantly lower than that for freshwater species (30.13ng TBTl(-1); lower confidence limit: 9.23ng TBTl(-1)), indicating that saltwater species are probably more susceptible to TBT than their freshwater counterparts.

Animals↗

The influence of nonylphenol on life-history of the earthworm Dendrobaena octaedra Savigny: linking effects from the individual- to the population-level.

We conducted a study to look at the effects of nonylphenol (NP) on the life-history of the parthenogenetic earthworm, Dendrobaena octaedra. During a 196-day study, we observed that the growth rate of juveniles and the percentage of worms producing cocoons were the only traits significantly affected by NP, while the total number of cocoons produced was marginally affected. Despite some fairly large changes in the average values of individual life-history traits caused by NP, the effects were difficult to detect statistically due to large interindividual variability. A declining trend was observed for population growth rate (lambda) with increasing NP concentration, but the decline was not statistically significant. The percent reduction in lambda was less than the percent reduction in the most sensitive life-history trait (fecundity). An elasticity analysis showed that lambda was more sensitive to changes in survival than to changes in reproductive traits. However, neither juvenile nor adult survival were affected by NP, and decomposition analysis showed that the minor changes in lambda were mainly caused by effects of NP on time to first reproduction, time between reproduction events and fecundity. The present study suggests that extrapolation from laboratory studies to population effects in the field may be greatly enhanced by combining ecotoxicological and demographic methods.

Animals↗

Effect of sublethal concentrations of chlorpyrifos on three successive generations of Daphnia carinata.

Effects of sublethal concentrations of chlorpyrifos (ranging from 0.005 (0.01 LC(50)) to 0.500 microg/L (1 LC(50))) on population characteristics of individual cultures of Daphnia carinata were investigated over 21 days with subsequent testing of the two next generations. The endpoints for the first and second generations observed were survival, fecundity, time to first brood, and number of offspring per female. The results were incorporated into the computation of the intrinsic rate of natural increase for daphnids in each of the treatments. Exposure to chlorpyrifos affected survival and fecundity of animals in the first generation. In the second generation, the most affected endpoint was time to the first brood with an indication of hormesis. The LC(50) tests were then conducted using animals of the third generation from each of the exposures in individual tests. Despite the absence of a negative effect of chlorpyrifos in the second generation, results of testing the third generation showed constant significant decline in LC(50) from control daphnids through to 0.1 LC(50) preexposed daphnids (0.1 LC(50), 0.05 microg/L, being the highest concentration in which animals survived exposure to the toxicant in the second generation).

Animals↗

The thyroid endocrine disruptor perchlorate affects reproduction, growth, and survival of mosquitofish.

The perchlorate anion--an oxidizer found in rockets, missiles, some ammunition, flares, airbags, and fireworks--occurs as a contaminant in ground and surface water in many parts of the United States. Its toxic effects include inhibition of thyroid hormone synthesis. To investigate its chronic toxicity, mosquitofish (Gambusia holbrooki) adults and fry were exposed to aqueous sodium perchlorate at 1, 10, and 100mg/L, and growth and reproductive performance (fecundity, eggs/embryos mass, and gonadosomatic index [GSI]) were determined. Five-day acute toxicity tests were also performed. Perchlorate had a stimulatory effect on fecundity, GSI, and egg/embryo mass, at least for some treatments. The LC50 of sodium perchlorate was 404 mg/L. Growth was enhanced at 1mg/L but inhibited at 10mg/L. These results suggest that, at environmentally relevant concentrations, perchlorate does not induce acutely toxic effects but may have mild stimulatory or hormetic effects on fitness parameters in this species.

Animals↗

Individual effects of estrogens on a marine fish, Cunner (Tautogolabrus adspersus), extrapolated to the population level.

Endocrine-disrupting chemicals (EDCs) in the environment may alter the population dynamics of wildlife by affecting reproductive output. This study describes a matrix modeling approach to link laboratory studies on endocrine disruption with potential ecological effects. The experimental model used is cunner (Tautogolabrus adspersus), which inhabit estuarine and marine areas where sewage treatment and other discharges containing estrogenic EDCs are likely. To test the effects of estrogenic exposures on fecundity, reproductively active cunner were exposed in three separate experiments by implanting 17beta-estradiol, estrone, or 17alpha-ethynylestradiol subcutaneously in a slow-release matrix at 0.05, 0.5, and 2.5 mg/kg. Egg production per gram female and egg viability were determined daily for a 1-week preexposure period and then for a 2-week exposure period. The mean number of eggs produced per gram female and egg viability (%) were calculated for the initial preexposure period and the 2-week exposure period for each experiment. Significant changes were observed in egg production per gram female in the high-17beta-estradiol treatment (P = 0.07) and high-17alpha-ethynylestradiol treatment (P = 0.04). A significant increase was observed in egg viability (%) in the low-17alpha-ethynylestradiol treatment (39.0%; P < or = 0.05). Cunner population response was projected using an age-structured matrix population model parameterized with published survival and fecundity estimates. By incorporating reproductive response data from laboratory exposures, model projections were used to describe how reproductive changes by estrogen treatment could alter cunner population growth rate (lambda). Published by Elsevier Inc.

Animals↗

Variation in the response of the invasive species Potamopyrgus antipodarum (Smith) to natural (cyanobacterial toxin) and anthropogenic (herbicide atrazine) stressors.

In the context of increasing freshwater pollution, the impact on life-traits (survival, growth and fecundity) and locomotion of Potamopyrgus antipodarum of a 5-week field-concentration exposure to the cyanobacterial toxin microcystin-LR and the triazine herbicide, atrazine was studied. Whatever the age of exposed snails (juveniles, subadults, adults), microcystin-LR induced a decrease in survival, growth and fecundity but had no effect on locomotion. Atrazine induced a decrease in locomotory activity but had no significant effect on the life-traits. These results are discussed in terms of consequences to field populations.

Animals↗

Polychlorinated biphenyls in serum and time to pregnancy.

Polychlorinated biphenyls (PCBs) may affect the female reproductive system in both animals and humans. In Sweden, a main exposure source to PCBs is consumption of fatty fish from the Baltic Sea. From 165 female consumers of such fish (fishermen's sisters), information on time to pregnancy (TTP) and miscarriages was collected, blood was drawn, and serum was analyzed for 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153), a biomarker for total PCB exposure. TTP and miscarriage data, as well as plasma CB-153 concentrations, for 121 fishermen's wives were available from previous studies. Thus, information on 286 women was available for investigation of whether a high exposure to PCB affects fertility negatively by increasing the TTP. The concentrations of CB-153 at the time of conception were estimated and trichotomized into low, medium, and high exposure groups. When we analyzed the joint data set of fishermen's wives and fishermen's sisters, a decrease rather than an increase in TTP was indicated for women in the medium (fecundability ratio 1.27 [95% confidence interval 0.89-1.82]) and high (fecundability ratio 1.42 [0.99-2.03]) exposure groups compared to those in the low exposure group. Similar results were found when we stratified on the original data set (fishermen's wives and fishermen's sisters) or childhood exposure. Women with miscarriages had lower estimated past CB-153 concentrations than women with live births. Our data provide no evidence of a hazardous effect associated with CB-153 concentration in the exposure range assessed. The exposure levels found in the fishermen's families, although high compared to that in the general Swedish population, may not be high enough to negatively affect fertility.

Abortion, Spontaneous↗

Effect of the exposure to Fasciola hepatica (Trematoda: Digenea) on life history traits of Lymnaea cousini and Lymnaea columella (Gastropoda: Lymnaeidae).

The snails Lymnaea columella and Lymnaea cousini have both been reported as intermediate hosts of Fasciola hepatica in Colombia. The effect of the exposure to the parasite on survival, fecundity and size of these snails was evaluated by means of experimental infections and the life history traits of control and exposed groups were compared. Infection rates were 82.2 and 34% for L. columella and L. cousini, respectively. A reduction in fitness was observed in both species when exposed to the parasite: fecundity alone was reduced in L. columella whereas in L. cousini there was also a decline in survival rate. Unlike other studies, increased size was not observed in either species. On the contrary, a reduction in growth rate was observed in L. columella.

Animals↗

A two-generation reproduction study to assess the effects of cows' milk on reproductive development in male and female rats.

OBJECTIVE: To examine whether the considerable quantities of increased female sex hormone levels found in modern milk as a result of modern dairy farming practices are safe for human consumption. DESIGN: Males and females of the P generation were maintained on a diet containing milk for 10 weeks before mating. Exposure to milk was continued up to the end of weaning of the F2b offspring. SETTING: Two-generation reproduction study. ANIMAL(S): Male and female Wistar Galas rats. INTERVENTION(S): P- and F1-generation rats were mated. MAIN OUTCOME MEASURE(S): Fertility, fecundity, and morphology and function of reproductive organs. RESULT(S): Although milk had growth-promoting effects in both parents and offspring, it caused no impairments in fertility, fecundity, or reproductive organ development in either generation. However, a whole litter from a dam of the P generation was born dead, three litters in total had a pup with skeletal abnormalities, and the AGD of F2a female pups was reduced. These events occurred only in the milk-treated rats. It is unknown whether these issues had any relevance to milk or only happened by chance. CONCLUSION(S): Further study is required to determine whether milk from pregnant cows is completely free from adverse effects on reproductive health.

Anal Canal↗

Larval helminths in intermediate hosts: does competition early in life determine the fitness of adult parasites?

Density-dependent effects on parasite fitness have been documented from adult helminths in their definitive hosts. There have, however, been no studies on the cost of sharing an intermediate host with other parasites in terms of reduced adult parasite fecundity. Even if larval parasites suffer a reduction in size, caused by crowding, virtually nothing is known about longer-lasting effects after transmission to the definitive host. This study is the first to use in vitro cultivation with feeding of adult trematodes to investigate how numbers of parasites in the intermediate host affect the size and fecundity of adult parasites. For this purpose, we examined two different infracommunities of parasites in crustacean hosts. Firstly, we used experimental infections of Maritrema novaezealandensis in the amphipod, Paracalliope novizealandiae, to investigate potential density-dependent effects in single-species infections. Secondly, we used the crab, Macrophthalmus hirtipes (Ocypodidae), naturally infected by the trematodes, M. novaezealandensis and Levinseniella sp., the acanthocephalan, Profilicollis spp., and an acuariid nematode. These four helminths all develop and grow in their crustacean host before transmission to their bird definitive host by predation. In experimental infections, we found an intensity-dependent establishment success, with a decrease in the success rate of cercariae developing into infective metacercariae with an increasing dose of cercariae applied to each amphipod. In natural infections, we found that M. novaezealandensis-metacercariae achieved a smaller volume, on average, when infrapopulations of this parasite were large. Small metacercariae produced small in vitro-adult worms, which in turn produced fewer eggs. Crowding effects in the intermediate host thus were expressed at the adult stage in spite of the worms being cultured in a nutrient-rich medium. Furthermore, excystment success and egg-production in M. novaezealandensis in naturally infected crabs were influenced by the number of co-occurring Profilicollis cystacanths, indicating interspecific interactions between the two species. Our results thus indicate that the infracommunity of larval helminths in their intermediate host is interactive and that any density-dependent effect in the intermediate host may have lasting effects on individual parasite fitness.

Acanthocephala↗