STOCHASTICITY, SCALE, AND REFRACTION: DETERMINANTS OF POPULATION SYNCHRONY IN PARASITES OF CENTRARCHIDS IN J. STROM THURMOND LAKE, SOUTH CAROLINA.

Among-deme asynchrony has the potential to influence community richness and diversity by increasing the likelihood of regional persistence for a species. Parasites of Lepomis spp. collected from 4 localities at J. Strom Thurmond Lake, South Carolina over a 1-yr period were used to evaluate patterns of parasite population synchrony. Localities were separated by approximately 5 km to increase the likelihood that the parasites sampled represented different demes. Tylodelphys scheuringi and Crinicleidus longus, exhibited negative covariation between synchrony and among-locality distances. The degree of synchrony exhibited by Neoechinorhynchus cylindratis, Crepidostomum cornutum, and Clavunculus bifurcatus was associated with the degree of similarity in habitat structure between localities. Patterns of synchrony for Posthodiplostomum minimum and Spinitectus sp. were not associated with any of the habitat variables examined. The influence of habitat structure on parasite population synchrony, possibly through the refraction of large-scale environmental drivers, has the potential to produce asynchronous dynamics that are independent of the distance between demes, thereby promoting regional persistence by increasing the likelihood of rescue effects.

QUANTIFYING BILATERAL INFECTION PATTERNS IN THE TREMATODE ALLOGLOSSIDIUM RENALE.

Within-host distributions of parasites can have relevance to parasite competition, parasite mating, transmission, and host health. We examined the within-host distribution of the adult trematode Alloglossidium renale infecting the paired antennal glands of grass shrimp. There are 4 possible parasite distributions for infections of paired organs: random, uniform, biased aggregation to 1 particular organ (e.g., left vs. right), or inconsistently biased (aggregated, but does not favor 1 side). Previous work has shown that morphological asymmetries in hosts can lead to biased infections of paired organs. Apparent symmetry between the antennal glands of grass shrimp leads to the prediction that there would be no bias for 1 particular organ. However, an alternative prediction stems from the fact that A. renale is hermaphroditic: aggregation between glands would increase outcrossing opportunities and thus, avoid inbreeding via self-mating. Existing methods to test for an overall pattern did not apply to the A. renale system because of low-intensity infections as well as many 0 values for abundance per unit of the antennal gland. Hence, we used Monte Carlo simulations to determine if the observed overall patterns differed from those expected by randomly allocating parasites into groups of 2. We found that in 3 of 4 data sets, A. renale infections did not deviate from random distributions. The fourth data set had a more uniform pattern than expected by chance. As there was no aggregation between glands and the proportion of worms in single gland infections did not differ from that expected by chance alone, we found no evidence of inbreeding avoidance as might be manifested via a within-host distribution. Given the large proportion of worms in single infections, we predict as a major evolutionary outcome that populations of A. renale will be largely inbred.

SAMPLE COMPOSITION: PARASITE ECOLOGY'S DIRTY LITTLE SECRET.

In comparative studies, the advantage of increased sample sizes might be outweighed by detrimental effects on sample homogeneity and comparability when small numbers of hosts from a different demographic of the same species are included in samples. A mixed sample of sunfishes (Lepomis spp.) was subdivided in different ways and examined using cumulative performance curves to determine whether the exclusion of larger hosts from a single-species sample and/or the inclusion of hosts of the same size demographic from closely related host species would produce more homogeneous samples. The exclusion of larger hosts from the single-species samples tended to reduce the aggregation of the infrapopulation samples, and mixed-species samples of smaller fishes tended to have lower degrees of aggregation for a given sample size relative to the single-species sample. Cumulative performance curves for diversity and richness, in concert with nonmetric multidimensional scaling of the infracommunities, demonstrated sunfish size to be a more reliable determinant of infracommunity similarity than sunfish species in this particular sample. The results demonstrate that cumulative aggregation curves can be an effective tool for delineating homogeneous and comparable subsamples and that, under some circumstances, it is possible to offset the smaller sample sizes that result from the exclusion of older/larger hosts by the addition of congeneric or confamilial hosts within the same size/age classes as the stratified sample.

Effect of Echinostoma caproni on Presumptive Lactic Acid Bacteria Abundance and Salmonella enterica Serovar Typhimurium Colonization in the Mouse Gut.

Co-infections of mammalian hosts with intestinal helminths and bacterial pathogens are common, especially in areas with inadequate sanitation. Interactions between co-infecting species and host microbiota can cause significant changes in host immunity, disease severity, and pathogen transmission, requiring unique treatment for each case. A greater understanding of the influences of parasite-bacteria co-infections will improve diagnosis and therapeutic approaches to control infectious diseases. To study the influence of the trematode parasite Echinostoma caproni on commensal and pathogenic bacteria in the mouse gut, we examined the abundance of intestinal lactic acid bacteria and Salmonella enterica serovar Typhimurium in control mice not exposed to E. caproni (P-) or S. Typhimurium (S-), E. caproni-infected (P+S-), S. Typhimurium-infected (P-S+), and E. caproni-S. Typhimurium co-infected (P+S+) mice, and determined bacterial burdens in the livers and spleens of the P-S+ and P+S+ mice. We also examined a subset of P+S- and P+S+ mice for survival and the relative location of E. caproni in the small intestine. The numbers of presumptive lactic acid bacteria were significantly higher in the P+S+ and P-S+ mice compared to the uninfected mice, and S. Typhimurium colonization in the liver and spleen was significantly reduced in the P+S+ mice compared to the P-S+ mice. Echinostoma caproni were located anteriorly in the intestine of P+S- mice, while in the P+S+ mice, the parasites were distributed more posteriorly. Survival of E. caproni was unaffected in either group. The results of our study suggest that E. caproni facilitates a higher abundance of presumptive lactic acid bacteria in the mouse intestine and reduces colonization of S. Typhimurium in the liver and spleen of the co-infected host.

Wrecking the Curve: Altered Functional Response of Tetragoneuria (Odonata: Corduliidae) Naiads Infected with Metacercariae of Haematoloechus floedae.

The ubiquity of host-parasite interactions and their potential for substantial representation, in terms of overall biomass, within ecosystems suggests that parasites have the capacity to influence energy flow within an ecosystem. Although the influence of certain parasites on prey behavior has been well documented, parasites could also exert an influence on ecosystem dynamics by influencing predator feeding behavior. The functional response of Tetragoneuria naiads was characterized by presenting naiads with varying abundances of Daphnia magna , after which a subset of the naiads were exposed to cercariae of Haematoloechus floedae, and the feeding trials repeated for both the control and exposed odonates. A type II functional response was chosen as an appropriate model for comparison. An indicator variable approach to nonlinear regression of the functional response data indicated that infected odonate naiads spent significantly more time foraging than they did before infection, whereas there was no significant change in the functional response of the control naiads. Infected odonates also had a slower rate of growth. These results imply a metabolic cost to infection of Tetragoneuria naiads by H. floedae that might be associated with the encapsulating response to the metacercariae that was observed in infected naiads.

Effect of Infection Duration on Habitat Selection and Morphology of Adult Echinostoma caproni (Digenea: Echinostomatidae) in ICR Mice.

The course of infection of Echinostoma caproni was followed in female ICR mice, a permissive laboratory host, from infection to natural termination. Twenty-one mice were infected with 20 metacercariae via oral intubation and housed 3 per cage. Three mice from a randomly selected cage were necropsied at 1 mo intervals. A second group of 15 mice was infected approximately 1 yr later to replace mice negative at necropsy in the first group. Mice in the second group were examined weekly for the presence of eggs in the feces. Mice negative for eggs on consecutive days were killed and necropsied. The location of individual worms and worm clusters were located in 20 segments of the small intestine. Adult worms were killed and fixed in hot formalin, stained, and prepared as whole mounts. Standard measurements were taken using a compound microscope fitted with an ocular micrometer. The infection spontaneously resolved in 10 mice from 7 to 32 wk PI, indicating the host response is highly variable and extending the maximum recorded length of E. caproni infections in ICR mice to 31 wk. A moribund worm was found in the feces of an animal that continued to pass eggs for an additional 2 mo indicating individual variation in worm responses. Worms located preferentially in the ileum (segments 11-13) during the first 3 mo of the infection but shifted to the jejunum (segments 8-9) during weeks 4-6. Morphologically, worms of different ages clustered together in multivariate space, with substantial overlap between the 3- and 4-mo-old infrapopulations and between the 5- and 6-mo-old infrapopulations. Muscular structures increased in size throughout the experiment, while the gonads increased in size for the first 3 mo and then declined during the last 3 mo. The relationship between E. caproni and ICR mice is more nuanced than previously reported. The reduction in gonad size and the shift from the ileum to the jejunum in the last 3 mo likely are related. These changes might be attributable to a localized immune response by mice to E. caproni that results in the ileum becoming less hospitable and a resultant relocation of the worms to a less favorable location in the jejunum.

Size, time, and asynchrony matter: the species-area relationship for parasites of freshwater fishes.

The tendency to attribute species-area relationships to "island biogeography" effectively bypasses the examination of specific mechanisms that act to structure parasite communities. Positive covariation between fish size and infrapopulation richness should not be examined within the typical extinction-based paradigm, but rather should be addressed from the standpoint of differences in colonization potential among individual hosts. Although most mechanisms producing the aforementioned pattern constitute some variation of passive sampling, the deterministic aspects of the accumulation of parasite individuals by fish hosts makes untenable the suggestion that infracommunities of freshwater fishes are stochastic assemblages. At the component community level, application of extinction-dependent mechanisms might be appropriate, given sufficient time for colonization, but these structuring forces likely act indirectly through their effects on the host community to increase the probability of parasite persistence. At all levels, the passive sampling hypothesis is a relevant null model. The tendency for mechanisms that produce species-area relationships to produce nested subset patterns means that for most systems, the passive sampling hypothesis can be addressed through the application of appropriate null models of nested subset structure.

Circadian egg production by Echinostoma caproni (Digenea: Echinostomatidae) in ICR mice.

Circadian egg production by Echinostoma caproni was investigated in ICR mice. Four female mice were infected with 25 E. caproni metacercariae, maintained in individual cages on a 12:12 light:dark cycle, and provided food and water ad libitum. Twenty-eight, 51, and 58 days post-infection, mice were transferred to individual, wire-bottomed cages and feces were collected every 2 hr for 24 hr. The feces were weighed and processed immediately to estimate the number of eggs present. Fecal output and egg production were standardized to unit maxima for analysis. Standardized egg count and standardized fecal output followed distinctly circadian patterns and covaried. Egg production was highest from 2200 to 0200 hr and lowest from 1000 to 1800 hr. These correspond to the highest and lowest fecal production, and highest and lowest periods of host activity, respectively. Egg density (eggs/g of feces) covaried weakly with fecal output with an additional peak at 0800-1000 hr, suggesting E. caproni is responding to changes in host physiology in timing of the production and release of eggs into the intestine. The continuous production and release of eggs during the patent period, coupled with the circadian pattern of daily egg release by E. caproni , would result in the widest dispersal of eggs in the host environment and enhance transmission to the first intermediate host.

Estimating prevalence: a confidence game.

Prevalence is one of the few estimates that rarely are reported with an appropriate measure of error in the parasitological literature. A minimum sample size recommendation of 15 samples, based on the relationship between sample size and standard error, likely has led to a false degree of confidence because of the nonlinear relationship between standard error and "true" 95% confidence intervals (as determined by Monte Carlo simulation or integration of the Bayesian posterior). Given that 95% confidence intervals for proportions are influenced by both sample size and the actual estimate of the proportion, there is no "gold standard" sample size beyond which estimates of binomial proportions can be considered "reliable." This necessitates the reporting of confidence interval estimates that have been shown to be conservative, such as the Clopper-Pearson estimate, or robust, such as the Wilson score approximation, or the computationally intensive integration of the Bayesian posterior.

Migratory response of Echinostoma caproni (Digenea: Echinostomatidae) to feeding by ICR mice.

The migratory response of Echinostoma caproni to host feeding was examined in female ICR mice. Thirty-six mice were each infected with 20 metacercariae of E. caproni . Twenty-eight days post-infection, food, but not water, was withheld for 24 hr. Mice were haphazardly divided into 4 groups of 9, and each group received one of the following treatments: (1) 0.25 g glucose, (2) access to standard lab chow, (3) 0.5 ml saline, and (4) continued fasting. Three mice from each treatment group were killed 1, 2, and 4 hr post-treatment. The intestine of each mouse was removed, flash-frozen, and stored in a conventional freezer for later examination. Intestines were partially thawed, measured, and opened longitudinally, and the position of each worm, or worm cluster was measured. The intestine was divided into equal 5% segments based on the initial measurement and locations of worms, and worm clusters were recorded from the appropriate section of the intestine for analysis. There was no significant effect of treatment in the position of worms at 1 hr. There was a posterior shift in worm position in all treatment groups at 2 hr, except in the saline-treated mice; however, only worms in the glucose-fed mice were significantly posterior to the unfed controls. From 2 to 4 hr, there was a significant anterior movement of worms in both the glucose and chow-fed mice. The data strongly suggest that E. caproni responds to the initiation of gastric activity of the host by migrating anteriorly in the ileum. The specific stimulus for this migration is unknown.

Parasites of flier, Centrarchus macropterus, from prairie and channel habitats in the Okefenokee Swamp, Georgia.

Flier (Centrarchus macropterus: Centrarchidae) were collected from a channel habitat, a prairie habitat, and a boundary between the 2 habitat types in March 2009 and examined for parasites. Flier from the prairie site had a significantly lower abundance of Pterocleidus acer and a significantly higher abundance of Hysterothylaceum juveniles. Patterns of infracommunity similarity showed a distinct break between the 2 habitat types, with fish collected from the boundary site clustering with either channel or prairie communities, suggesting that the small home range of flier restricts the exposure of individuals to parasites.

Examining the area effect for parasite communities of bluegill x green sunfish hybrids in five constructed ponds in Kansas.

The parasite communities of bluegill x green sunfish hybrids were examined from 5 constructed ponds in Kansas in an attempt to evaluate the separate effects of habitat area and habitat heterogeneity on parasite community structure. Characterization of fish community structure and collection of hybrid fishes was conducted using an electrofishing boat. Benthic invertebrates were sampled, and substrate types examined at 30 evenly spaced points in each pond. Nonmetric multidimensional scaling of the parasite infracommunities, in concert with an analysis of similarities, indicated significant clustering of infracommunities by locality. The number and diversity of habitat types, and the richness and diversity of both fishes and benthic invertebrates, were positively correlated with the first axis of the infracommunity ordination. Pond surface area, parasite richness, and stocking pressure were negatively correlated with the first axis of the infracommunity ordination, suggesting that pond area, stocking pressure, or both was a stronger determinant of parasite community structure in these systems than habitat and host heterogeneity.

Diurnal migration of Echinostoma caproni (Digenea: Echinostomatidae) in ICR mice.

Twenty-four female ICR mice, 12 acclimated to a 12 ∶ 12 light-dark cycle and 12 to a 12 ∶ 12 dark-light cycle for 7 days, were each infected with 10 metacercariae of Echinostoma caproni. Infected mice were maintained on their respective lighting regimes for 28 days. Six mice (3 from each group) were necropsied at 4-hr intervals beginning at 0700 hr. The small intestine was removed, opened, and the position of individual worms and worm clusters was measured to the nearest 0.1 cm. Each intestine was subsequently divided into 20 equal segments and individual worms and worm clusters were assigned to the appropriate segment based on the original measurements. All worms were found in the posterior 55% of the intestine (ileum). All posterior segments (10-20), with the exception of segment 18, harbored at least 1 worm at some time. A Monte Carlo simulation of worm abundance in segments 10-17 over all time periods indicated a random distribution, while the same analysis of segments 10-20 indicated a non-random distribution due to large numbers of worms in segment 20 and to the absence of worms in segment 18. To analyze temporal changes in worm distribution, mice were grouped by time of necropsy as follows: night (1900 and 2300 hr), morning (0300 and 0700 hr), and day (1100 and 1500 hr). During the night and morning, E. caproni was heavily concentrated in segments 10-17 and, during the day, worms were located more posteriorly, with a heavy concentration in the last segment (20).

Prerequisites for parasitism in rhabditid nematodes.

To evaluate their potential for survival in a vertebrate host, dauer larvae from 7 species of rhabditid nematodes were subjected to in vitro conditions designed to emulate those of a vertebrate digestive tract. Dauer larvae from 3 of the 7 species, selected for their ability to survive elevated temperatures and low pH, and representing differing types of phoretic associations with invertebrate hosts, were fed to frogs to examine their ability to survive passage through a vertebrate digestive system. The degree of invasiveness of the phoretic association that dauer larvae had with their invertebrate hosts did not correspond to patterns of in vitro survivorship for any of the experimental conditions. When consumed with a prey item, dauer larvae from all 3 species were recovered from frogs 72 hr postexposure, and no differences for in vivo survivorship were observed among the 3 species. The contention that invasiveness or facultative parasitism within an invertebrate host is a beneficial or necessary step toward vertebrate parasitism by rhabditid nematodes was not supported by the survivorship data.

The interaction of light and gravity on the transmission of Echinostoma caproni (Digenea: Echinostomatidae) cercariae to the second intermediate host, Biomphalaria glabrata (Gastropoda: Pulmonata).

The current experiments were designed to assess the interaction of light and gravity on the transmission of Echinostoma caproni cercariae to the second intermediate host, Biomphalaria glabrata. Transmission chambers were constructed of clear polyvinyl chloride pipe covered with a black sleeve to exclude light. Snails were constrained within the chamber to prevent movement, while permitting the cercariae to swim freely. A trial consisted of 2 infected B. glabrata shedding E. caproni cercariae placed at the center of the chamber with 5 uninfected B. glabrata placed 10 cm above and below the shedding snails as sentinels. Three experiments, consisting of 12 trials each, were conducted under the following lighting conditions, i.e., above and below the transmission chamber, and in complete darkness. In all 3 experiments, the proportion of metacercariae was significantly higher in snails at the top of the chamber. The results suggest that a negative geotaxis is the primary factor in the initial dispersal of E. caproni cercariae. Coupling negative geotaxis and positive phototaxis (light from above) resulted in a significantly higher proportion of metacercariae in sentinel snails at the top of the transmission chamber when corrected for cercarial density. There was no significant difference in the proportion of metacercariae in snails at the top or bottom of the transmission chamber with light at the bottom of the chamber or in complete darkness. Cercariae of E. caproni only respond to light in context, i.e., from above, and ignore the light stimulus when it comes from an unexpected location (bottom of the water column). Significantly greater numbers of cercariae were released from shedding snails when light was present, suggesting that emergence of cercariae from B. glabrata is dependent on light regardless of the position of the light source.

Helminth infracommunities of the common snapping turtle (Chelydra serpentina serpentina) from Westhampton Lake, Virginia.

Patterns of infracommunity similarity were examined for 27 male and 6 female common snapping turtles, Chelydra serpentina serpentina, collected from Westhampton Lake on the campus of the University of Richmond in Richmond, Virginia, during the summer months of 1979 and 1980. Patterns of infracommunity similarity based on parasite abundance emphasized differences between years and between host sexes. Patterns of similarity based on parasite presence or absence emphasized differences among the months sampled. This suggests that there were consistent seasonal changes across both years in terms of which parasites were present, but that there were differences between years in terms of the abundances of those parasites.

"Active" passive sampling in two species of Lepomis from Par Pond, South Carolina, United States: a case study of infracommunity nestedness.

The influence of ontological diet shifts on infracommunity nestedness was examined by comparing the infracommunity structure of Lepomis gulosus and Lepomis macrochirus at 2 localities in Par Pond, South Carolina, United States. Fill-constrained, occurrence-constrained, and abundance-constrained null models were used to evaluate the degree of nestedness. The presence-absence matrices from all 4 component communities had significantly fewer discrepancies than those produced by the fill-constrained model, and none had significantly fewer discrepancies than those produced by the occurrence-constrained model. Only the presence-absence matrix for the infracommunities of L. gulosus from the Cold Dam locality had significantly fewer discrepancies than those produced by the abundance-constrained null model. The nestedness of the 4 samples could not be distinguished from that expected under a hypothesis of passive sampling. A positive correlation between host size and total parasite abundance indicates the passive mechanism has a deterministic basis. Thus, even in the absence of habitat or diet shifts, nestedness can arise in infracommunities of freshwater fishes when older, larger fish have sampled more individuals from an uneven distribution of infective stages.

Structure and similarity of helminth communities of six species of Australian turtles.

Patterns of infracommunity structure and infra- and component community similarity were examined for helminths of 6 species of turtles, each collected from a single locality in Australia in 1993 and 1994. Elseya latisternum (N = 11) and Emydura kreffti (N = 16) were collected from northern Queensland, Emydura macquarii macquarii (N = 11) from southern Queensland, Emydura macquarii dhara (N = 11) and Chelodina longicollis (N = 11) from northern New South Wales, and Chelodina oblonga (N = 5) from Western Australia. Local parasite species richness was not correlated with host geographical range. Differences in parasite diversity among host species were related primarily to differences in evenness, a pattern attributed to local habitat characteristics, rather than species-specific differences in colonization potential. Ordination and analysis of similarity demonstrated the patterns of infracommunity structure of Chelodina spp. to be distinct from those of the other host species sampled, which showed considerable overlap among patterns of infracommunity structure. Despite overlap with the component communities of Em. kreffti and El. latisternum, the component communities of Em. m. dhara and Em. m. macquarii were more distinct from one another than either was to the component communities of Em. kreffti or El. latisternum.

Nestedness in colonization-dominated systems: helminth infracommunities of Rana vaillanti Brocchi (Anura: Ranidae) in Los Tuxtlas, Veracruz, Mexico.

Colonization probabilities of parasite species often are determined by the habitat preference and vagility of host individuals. Although extinction-based interpretations have been investigated for nested subset patterns of parasite infracommunities, the low relative frequency of nestedness in colonization-dominated systems makes the determination and interpretation of nested infracommunities of broad ecological importance. In these systems, ontogenetic shifts in habitat preference or diet of the host have the potential to produce nested subset patterns of parasite infracommunities. Helminth infracommunity structure was investigated for 76 Rana vaillanti individuals collected from Laguna Escondida, Los Tuxtlas, Veracruz, Mexico, in 1998. Pooled helminth infracommunities were significantly nested, as were penetrating and ingested helminth infracommunities when considered separately. Richness, diversity, and evenness of the helminth infracommunities were not correlated with host size, and did not differ between host sexes, suggesting that the structure of infracommunities simply is a product of the interaction between host individuals and their landscape mediated by individual differences in vagility. It is hypothesized that individual differences in recruitment can produce nested subset infracommunity patterns when the habitats or habitat preferences of hosts are themselves nested.