Living on the edge: comparative phylogeography and phylogenetics of Oreohelix land snails at their range edge in Western Canada.

BACKGROUND: The biodiversity and distributions of terrestrial snails at local and regional scales are influenced by their low vagility and microhabitat specificity. The accessibility of large-bodied species and their characteristically high levels of genetic polymorphism make them excellent ecological and evolutionary models for studies on the phylogeography, phylogenetics, and conservation of organisms in fragmented populations. This study aims to elucidate the biodiversity, systematics, and distributions of genetic lineages within the genus Oreohelix at the northern and western periphery of their range. RESULTS: We found four mitochondrial clades, three of which are putative subspecies of Oreohelix subrudis. One clade was geographically widespread, occurring within numerous sites in Cypress Hills and in the Rocky Mountains, a second was geographically restricted to the Rocky Mountains in Alberta, and a third was restricted to the Cypress Hills region. A fourth clade was the small-bodied species, O. cooperi. ITS2 sequence and screening data revealed three genetic clusters, of which one was O. cooperi. Cluster 1 contained most individuals in COI clade X and some from clade B and cluster 2 was predominantly made up of individuals from COI clades B and B' and a few from clade X. ITS2 alleles were shared in a narrow contact zone between two COI clades, suggestive of hybridization between the two. CONCLUSIONS: A sky island known as Cypress Hills, in southeastern Alberta, Canada, is a biodiversity hotspot for terrestrial land snails in the genus Oreohelix. The observed phylogeographic patterns likely reflect reproductive isolation during the Last Glacial Maximum, followed by secondary contact due to passive, long-range dispersal resulting from low vagility, local adaptation, and complex glacial history.

Parasite-induced oxidative stress in liver tissue of fathead minnows exposed to trematode cercariae.

Although results from field surveys have linked parasites to oxidative stress in their fish hosts, direct evidence involving experimentally infected hosts is lacking. We evaluated the effects of experimental infections with larval trematodes on induction of oxidative stress in fathead minnows, Pimephales promelas. Juvenile fish were exposed in the laboratory to the larvae (cercariae) of 2 species of trematode: Ornithodiplostomum sp. that develops in the liver, and O. ptychocheilus that develops in the brain. For Ornithodiplostomum sp., lipid peroxidation concentration in liver tissue increased 5 days after exposure and remained higher than controls until the end of the experiment at 28 days. For O. ptychocheilus, liver lipid peroxidation concentration was higher than controls at 5 days, but not thereafter. Sustained elevation in lipid peroxidation concentration for the liver trematode may be explained by direct tissue damage caused by developing larvae in the liver, or by an immune response. These experimental results support those from field studies, indicating that the lipid peroxidation assay may be an effective biomonitor for parasite-induced oxidative stress in fish, and that the nature of the oxidative stress response is species and/or tissue specific.

Evaluation of diagnostic techniques for Anoplocephala perfoliata in horses from Alberta, Canada.

Accurate diagnosis of parasites within individual hosts remains a difficult task. Incorrect and/or inaccurate diagnosis restricts the potential for targeted treatment of individuals and limits our understanding of key epidemiological characteristics of potential pathogens of domestic stock. In this study, we compared the specificity and sensitivity of four diagnostic methods for determination of the presence and intensity of the cestode Anoplocephala perfoliata in horses. Over 400 horses from an abattoir in south-central Alberta were evaluated for the presence and number of cestodes. Thirty one horses were infected with 1 to >1000 worms. Diagnosis based upon faecal egg counts of horses with known numbers of worms was least accurate in detecting worm presence. Detection of circulating antibodies to the cestode was most sensitive using Western blot analysis (100%), but had lower specificity (87%). A serum-based ELISA had a lower sensitivity (70%) for detection of antibodies. A coproantigen ELISA had 74% sensitivity and 92% specificity, and there was a positive correlation between antigen concentration and tapeworm intensity. The coproantigen ELISA may have the potential as a diagnostic tool for determining the presence and intensity of this potentially pathogenic cestode.

Physico-chemical determinants of helminth component community structure in whitefish (Coregonus clupeaformes) from adjacent lakes in Northern Alberta, Canada.

Populations of hosts vary extensively in the types and numbers of parasites that the average individual contains. Understanding the factors that lead to this variation is an important goal for parasite ecologists. We characterized patterns of helminth component community structure in whitefish collected from a cluster of 7 lakes located on an isolated plateau in northern Alberta, Canada. Component communities were species rich (5-6 species per lake), high in mean helminth intensity (approximately 80-500 individuals/host), and high in between-lake similarity (50-100%), a pattern consistent with results from studies on whitefish sampled from other localities in Northern Canada and Europe. Multivariate analyses indicated that the structure of the component communities was associated with 2 opposing environmental gradients. One was defined primarily by water colour, the second by phosphorous concentration. Thus, 4 lakes were characterized by a combination of high colour, low productivity, low parasite intensities, and the absence of larval acanthocephalans. Habitat/species associations were less clear as intensities increased, but the 3 remaining lakes tended to have the opposite characteristics. These results provide evidence that variation in helminth component community structure in fish is associated with variation in physicochemical characteristics that are linked to aquatic productivity.

Chronology of parasite-induced alteration of fish behaviour: effects of parasite maturation and host experience.

We monitored temporal changes in the magnitude of altered host behaviour in minnows (Pimephales promelas) experimentally infected with metacercariae of a brain-encysting trematode (Ornithodiplostomum ptychocheilus). This parasite develops and then encysts in a region of the brain that mediates the optomotor response (OMR), an innate behaviour that links visual stimuli with motor performance. The OMR of infected and uninfected minnows was evaluated between 0 and 10 weeks post-infection (p.i.), an interval spanning the development period of metacercariae to infectivity in birds. Trials involved monitoring the time an individual minnow spent following a spinning drum that had been painted with alternating black and white stripes. At 2 and 4 weeks p.i., infected minnows followed the drum 40% less often than controls. Differences between controls and infected fish declined thereafter, and were undetectable by 10 weeks p.i. Both control and infected fish habituated equally rapidly to the spinning drum. However, the difference in performance between controls and infected fish was 29% for experienced fish and 48% for fish that had never experienced the drum. Because maximum parasite-induced reduction in OMR coincided with the period of maximum parasite development, the behavioural effects are most likely due to unavoidable pathology in the brain associated with developing larvae.

Population dynamics of Ornithodiplostomum ptychocheilus metacercariae in fathead minnows (Pimephales promelas) from four northern-Alberta lakes.

Annual, seasonal, and interlake variation in prevalence and intensity of Ornithodiplostomum ptychocheilus (Faust) metacercariae was assessed in populations of fathead minnows (Pimephales promelas) collected from 4 lakes in north-central Alberta. Mean metacercariae intensity in young-of-the-year minnows varied extensively (5-123 metacercariae/host) among year, month, and lakes. In 2 of the lakes, prevalence always reached 100%, and mean intensity always peaked in September or October. The high spatial and annual variation in metacercarial recruitment was partly attributable to variation in host size. but variation in water depth, temperature, snail densities, and bird visitation likely also played a role. A laboratory experiment demonstrated that host and metacercariae survival was intensity-independent during a period of simulated winter. Thus, metacercariae recruited in the fall survive until the following spring.

Parasite-induced variation in host morphology: brain-encysting trematodes in fathead minnows.

Metacercariae of the trematode Ornithodiplostomum ptychocheilus cause a conspicuous enlargement of the cranium of juvenile fathead minnows (Pimephales promelas). Minnows sampled from 2 naturally infected ponds in northern Alberta, Canada, had 12% higher and 7% wider craniums compared to fish from an adjacent, uninfected pond. We tested the prediction that cranial distortion was caused by encystment of metacercariae on the brains of slow-growing minnows in a factorial experiment. Juvenile fish were either exposed once to 120 cercariae or 3 times to 40 cercariae; they were then fed either a low- or high-quantity diet for 8 wk. Results showed that after controlling for host size, cranial heights were affected by infection regime and host diet but not by the infection x diet interaction. Cranial distortion was most prominent in minnows exposed once to cercariae, showing that the rapid, simultaneous growth of metacercariae interfered with the normal development of the cranium. Thus, the expression of the parasite-induced phenotype was context dependent, the result of factors associated with the dynamics of cercariae transmission and host growth rate.

Development and intensity dependence of Ornithodiplostomum ptychocheilus metacercariae in fathead minnows (Pimephales promelas).

Intensity-dependent development of Ornithodiplostomum ptychocheilus metacercariae was studied in fathead minnows (Pimephales promelas) exposed to 0, 20, or 120 cercariae. Subsamples of hosts were necropsied at 2-wk intervals to monitor parasite recruitment, growth, and time to encystment. The complex development of metacercariae within the cranium of minnows involved growth, encystment, and consolidation phases, each of which were affected by intensity. At the end of the growth phase, metacercariae from low-dose fish were 20% longer than those from high-dose fish and the latter took 2-4 wk longer to encyst. At the end of the postencystment consolidation phase (6-8 wk postinfection), the size of metacercariae decreased by approximately 50%. The rate of consolidation was slower in high-dose fish. Our results show that time, intensity, and temperature affect development of O. ptychocheilus. Because metacercariae development and differentiation are linked to infectivity, events occurring in intermediate hosts can potentially impact the structure and size of trematode suprapopulations.

Experimental population dynamics of Rhabdias bufonis (Nematoda) in toads (Bufo bufo): density-dependence in the primary infection.

Density-dependence in worm establishment, numbers, biomass and larval production were examined in primary infections of 0, 10, 40, 80 and 160 larvae of the lung nematode, Rhabdias bufonis in the common toad, Bufo bufo. The infection procedure established 4 non-overlapping levels of infection which persisted until 6 weeks post-infection (p.i.), after which there was an overall decline up to 12 weeks p.i. Worm numbers had no direct effect on adult worm survival but temporal changes in worm weight were density-dependent. Adult worm establishment in the lungs declined significantly as the numbers of worms in the lungs increased. At the lowest exposure dose, 86% of the larvae administered reached maturity in the lungs while at the highest, only 37% did so. Also, the numbers of immature larvae outside the lungs increased as adult worm numbers increased. Both features provide evidence for a threshold limit to the numbers of worms maturing in the lungs. Worm numbers also affected larval output per host and per capita fecundity. A significant positive relationship between per capita fecundity and per capita worm weight suggested that density-dependence acted primarily to constrain the growth of individual worms. Finally, the constraints imposed on worm growth and fecundity were apparently relaxed when worm density decreased, providing evidence for density-dependent flexibility in per capita fecundity. Density-dependence in worm establishment and per capita fecundity are mechanisms which may potentially regulate this host-parasite interaction in the field. Both mechanisms may be functionally related to physical space limitations in the lungs, within which worms must compete for finite nutrients.

Negative effects of Rhabdias bufonis (Nematoda) on the growth and survival of toads (Bufo bufo).

The growth and survival of juvenile toads, Bufo bufo, infected with a common lung nematode, Rhabdias bufonis, were studied. Toads were raised from tadpoles in the laboratory and infected 2 months after metamorphosis. Individual toads were exposed to doses of 10, 40, 80 or 160 larvae, which enabled examination of the hypothesis that parasite-induced mortality is affected by worm numbers. Growth of infected toads began to diverge from that of uninfected controls at 6 weeks post infection (WPI) and by 12 WPI the most heavily infected toads were approximately half the mass of controls. No controls died throughout the experiment; however, mortality of infected toads was strongly affected by parasite density. A mechanism for mortality is suggested by the significant negative relationship between parasite density and dietary intake. This parasite-induced anorexia was detected at 3 WPI and persisted up to 9 WPI. Patterns of reduced host growth, survival and dietary intake provide experimental evidence of the negative consequences of parasitic infection in a natural parasite-host system which may also be present under natural conditions.