Description, life cycle, and development of the myxozoan Myxobolus rasmusseni n. sp. in fathead minnows, Pimephales promelas: A possible emerging pathogen in southern Alberta, Canada.

Morphological, gene sequence, host tissue tropism, and life cycle characteristics were utilized to describe the myxozoan, Myxobolus rasmusseni n. sp. from fathead minnow, Pimephales promelas, collected from reservoirs in southern Alberta. Results from serial histological sections of whole heads showed that myxospores were contained within irregular-shaped and sized coelozoic capsules (=plasmodia). Clusters of membrane-bound, myxospore-filled plasmodia filled the head cavities of juvenile fathead minnows, leading to the development of large, white, disfiguring lesions in mid to late summer. Bilateral exopthalmia (pop-eye disease) was a common outcome of M. rasmusseni n. sp. development. BLASTn search of a 1974 bp sequence of the 18S rDNA gene isolated from myxospores indicated that M. rasmusseni n. sp. was distinct from other coelozoic and histozoic Myxobolus spp. cataloged in GenBank. 18S rDNA gene sequences from triactinomyxon spores released from the oligochaete Tubifex were 100% identical to sequences from myxospores collected from syntopic fathead minnows. Results from a longitudinal survey of the 2020 cohort of fathead minnows showed that young-of-the-year are exposed at 1-5 mo and that 60-90% of these had developed myxospore-filled lesions approximately one year later. Data regarding potential sources and timing of M. rasmusseni n. sp. emergence in fathead minnow populations are needed.

A congeneric and non-randomly associated pair of larval trematodes dominates the assemblage of co-infecting parasites in fathead minnows (Pimephales promelas).

Individual hosts are often co-infected with multiple parasite species. Evidence from theoretical and empirical studies supports the idea that co-occurring parasites can impact each other and their hosts via synergistic or antagonistic interactions. The fundamental aim of understanding the consequences of co-infection to hosts and parasites requires an understanding of patterns of species co-occurrence within samples of hosts. We censused parasite assemblages in 755 adult, male fathead minnows collected from 7 lakes/ponds in southern Alberta, Canada between 2018 and 2020. Fifteen species of endoparasites infected fathead minnows, 98% of which were co-infected with between 2 and 9 parasite species (mean species richness: 4.4 ± 1.4). Non-random pairwise associations were detected within the overall parasite community. There were particularly strong, positive associations in the occurrences and intensities of the 2 congeneric larval trematodes Ornithodiplostomum sp. and Ornithodiplostomum ptychocheilus that comprised >96% of the 100 000+ parasites counted in the total sample of minnows. Furthermore, the occurrence of Ornithodiplostomum sp. was a strong predictor of the occurrence of O. ptychocheilus, and vice versa. Positive covariation in the intensities of these 2 dominants likely arises from their shared use of physid snails as first intermediate hosts in these waterbodies. These 2 species represent a predictable and non-random component within the complex assemblage of parasites of fathead minnows in this region.

TREMATODE CLONE ABUNDANCE DISTRIBUTIONS: AN ECO-EVOLUTIONARY LINK BETWEEN PARASITE TRANSMISSION AND PARASITE MATING SYSTEMS.

Most trematodes and some cestodes have obligate life history features that include an asexual developmental stage that can produce genetically-identical individuals (clonemates) followed by an adult stage with sexual reproduction. These life history features can influence the evolutionary mechanism of inbreeding in parasites, especially among self-compatible hermaphroditic endoparasites whose mating opportunities are restricted to within hosts. As clonemate mating in hermaphroditic species produces a genetic inbreeding signature identical to that of self-mating, it is important to understand how clonemates are transmitted through their life stages. A handful of prior studies compared clone richness (number of clones) across life cycle stages to infer transmission processes and to characterize clone abundance distributions (CADs) among hosts. Here we illustrate the use of the proportion of clonemate dyads (PC) within hosts to describe the CADs. PC has several advantages as an ecological metric in that it is unbiased by sample size, takes into account relative parasite burdens, and has a direct transmission interpretation, i.e., the probability of cotransmitting clonemates. Moreover, PC is also an evolutionary metric as it can be used to estimate a potential clonemate mating rate. We demonstrate the use of PC in comparing CADs within and across 2 trematode developmental stages in the lancet fluke Dicrocoelium dendriticum. Also, we show how genetic estimates of apparent selfing (true selfing plus clonemate mating) at larval and juvenile stages can be compared to PC estimated at the adult stage to assess the contribution of clonemate mating to apparent selfing. The eco-evolutionary links presented are generalizable to assess sibling cotransmission as well. Thus, the framework presented herein will facilitate future field-based studies on the transmission and mating systems of parasitic flatworms.

Nonhost species reduce parasite infection in a focal host species within experimental fish communities.

The dilution effect describes the negative association between host biodiversity and the risk of infectious disease. Tests designed to understand the relative roles of host species richness, host species identity, and rates of exposure within experimental host communities would help resolve ongoing contention regarding the importance and generality of dilution effects. We exposed fathead minnows to infective larvae of the trematode, Ornithodiplostomum ptychocheilus in minnow-only containers and in mixed containers that held 1-3 other species of fish. Parasite infection was estimated as the number of encysted worms (i.e., brainworms) present in minnows following exposure. The results of exposure trials showed that nonminnow fish species were incompatible with O. ptychocheilus larvae. There was no reduction in mean brainworm counts in minnows in mixed containers with brook sticklebacks or longnose dace. In contrast, brainworm counts in minnows declined by 51% and 27% in mesocosms and aquaria, respectively, when they co-occurred with emerald shiners. Dilution within minnow + shiner containers may arise from shiner-induced alterations in minnow or parasite behaviors that reduced encounter rates between minnows and parasite larvae. Alternatively, shiners may act as parasite sinks for parasite larvae. These results highlight the role of host species identity in the dilution effect. Our results also emphasize the complex and idiosyncratic effects of host community composition on rates of parasite infection within contemporary host communities that contain combinations of introduced and native species.

Clonemate cotransmission supports a role for kin selection in a puppeteer parasite.

Host manipulation by parasites is a fascinating evolutionary outcome, but adaptive scenarios that often accompany even iconic examples in this popular field of study are speculative. Kin selection has been invoked as a means of explaining the evolution of an altruistic-based, host-manipulating behavior caused by larvae of the lancet fluke Dicrocoelium dendriticum in ants. Specifically, cotransmission of larval clonemates from a snail first host to an ant second host is presumed to lead to a puppeteer parasite in the ant's brain that has clonemates in the ant abdomen. Clonal relatedness between the actor (brain fluke) and recipients (abdomen flukes) enables kin selection of the parasite's host-manipulating trait, which facilitates transmission of the recipients to the final host. However, the hypothesis that asexual reproduction in the snail leads to a high abundance of clonemates in the same ant is untested. Clonal relationships between the manipulator in the brain and the nonmanipulators in the abdomen are also untested. We provide empirical data on the lancet fluke's clonal diversity within its ant host. In stark contrast to other trematodes, which do not exhibit the same host-manipulating behavioral trait, the lancet fluke has a high abundance of clonemates. Moreover, our data support existing theory that indicates that the altruistic behavior can evolve even in the presence of multiple clones within the same ant host. Importantly, our analyses conclusively show clonemate cotransmission into ants, and, as such, we find support for kin selection to drive the evolution and maintenance of this iconic host manipulation.

Spatiotemporal Patterns of Infection for Emerging Larval Liver Fluke ( Dicrocoelium dendriticum) in Three Species of Land Snail in Southern Alberta, Canada.

The control of emerging parasites requires a fundamental knowledge of where and when rates of transmission are high. Data on spatiotemporal patterns of infection are challenging to obtain, particularly for complex life cycle parasites that involve transmission into multiple obligate hosts. The lancet liver fluke, Dicrocoelium dendriticum, has a long history of colonization outside its native host and geographical range in continental Europe. Infection patterns involving adult and metacercarial stages have been characterized for this trematode in a region of emergence in western Canada within co-grazing herbivores and ants, but infection patterns in snail intermediate hosts in this region are unknown. We combined spatiotemporal prevalence surveys with sequence analyses of the cytochrome c oxidase subunit 1 ( COI) barcoding gene from samples of sporocyst tissue in infected snails to confirm that D. dendriticum utilizes 3 sympatric species of Oreohelid land snail ( Oreohelix subrudis, Oreohelix sp., and Oreohelix cooperi) as first intermediate host. Mean prevalence within a total sample of 900 adult snails collected over 1 field season from 6 sites was 9.9 ± 2.4%. For each species of snail, prevalence ranged between 5-30% within monthly samples, with peaks in mid-summer followed by declines in fall. Between-site variation in prevalence was low and non-significant, implying that rates of transmission of D. dendriticum miracidia from domestic stock and wildlife into snails are similar within localized sites, despite high variation in local habitat characteristics and in the structure of the definitive host community.

3D virtual histology at the host/parasite interface: visualisation of the master manipulator, Dicrocoelium dendriticum, in the brain of its ant host.

Some parasites are able to manipulate the behaviour of their hosts to their own advantage. One of the most well-established textbook examples of host manipulation is that of the trematode Dicrocoelium dendriticum on ants, its second intermediate host. Infected ants harbour encysted metacercariae in the gaster and a non-encysted metacercaria in the suboesophageal ganglion (SOG); however, the mechanisms that D. dendriticum uses to manipulate the ant behaviour remain unknown, partly because of a lack of a proper and direct visualisation of the physical interface between the parasite and the ant brain tissue. Here we provide new insights into the potential mechanisms that this iconic manipulator uses to alter its host's behaviour by characterising the interface between D. dendriticum and the ant tissues with the use of non-invasive micro-CT scanning. For the first time, we show that there is a physical contact between the parasite and the ant brain tissue at the anteriormost part of the SOG, including in a case of multiple brain infection where only the parasite lodged in the most anterior part of the SOG was in contact with the ant brain tissue. We demonstrate the potential of micro-CT to further understand other parasite/host systems in parasitological research.

Hyperparasitism of an Avian Ectoparasitic Hippoboscid Fly, Ornithomya anchineuria, by the Mite, Myialges Cf. Borealis, in Alberta, Canada.

Hippoboscid flies (Diptera: Hippoboscidae) include species that are ectoparasites of birds in the Northern Hemisphere, but little is known regarding their taxonomy, parasites, avian host associations, or geographical distribution in North America. In late August of 2013 and 2014, we collected hippoboscid flies from live birds trapped in mist nets as part of a banding study in Cypress Hills Interprovincial Park in southeastern Alberta, Canada. A total of 113 birds comprising 9 species was examined in 2013. Of these, 18 individuals were infested with 1-3 Ornithomya anchineuria Speiser (n = 22 flies; prevalence = 15.9%). Eight of these flies carried 1-8 adult female epidermoptid mites anchored to their ventral, posterior abdomens. Each female was associated with clusters of up to 30 stalked eggs. The first pair of tarsi on adult female mites was highly modified as anchors, indicating permanent attachment through the host cuticle. Morphological traits identified these mites as Myialges cf. borealis Mironov, Skirnisson, Thorarinsdottier and Nielsen. Cytochrome c oxidase subunit 1 ( COX1) gene sequences obtained for 2 mites were distinct from those previously reported for species of Myialges, being most similar to Myialges trinotoni Cooreman. The paucity of available gene sequences for Myialges and related genera of epidermoptid mites prevents any further conclusions regarding taxonomy. These findings extend previous reports of O. anchineuria from Pacific and Atlantic coasts of Canada inland to the central migratory flyway of the Northern Great Plains and expand the limited information available for Myialges spp.

User-friendly Taqman probe coupled-insulated isothermal PCR (iiPCR) for rapid detection of emerging Ambystoma tigrinum virus (ATV) in western tiger salamanders (Ambystoma mavortium) on a compact, portable instrument.

Portable user-friendly diagnostic tests can benefit detection and surveillance of wildlife diseases. Here, the performance of a compact POCKIT™ Nucleic Acid Analyzer for detection of Ambystoma tigrinum virus (ATV), an emerging Iridovirus that is associated with high host mortality in the western tiger salamander (Ambystoma mavortium) in North America was assessed. Tissue samples from 188 larval tiger salamanders collected from sites in Alberta, Canada were tested by both iiPCR and by conventional PCR. Results of the two assays showed 96.3% agreement. All 176 samples that tested positive by conventional PCR were also positive by iiPCR, while 12 of the samples that were negative by conventional PCR were positive by iiPCR. Comparison of the limits of detection of the two assays shows that the iiPCR assay was more sensitive than conventional PCR and had a LOD95 of 20 copies per reaction. The instrument automatically analyzes and displays results within 40min following nucleic acid extraction. The novel technology could enhance detection of, and response to, wildlife pathogens, particularly those that occur sporadically, cause rapid outbreaks, and/or occur within isolated geographical regions.

Population genetic analysis informs the invasion history of the emerging trematode Dicrocoelium dendriticum into Canada.

Parasite distributions are constantly changing due to climate change, local and global movement of animals and humans, as well as land use and habitat change. The trematode Dicrocoelium dendriticum is a relatively recent invader of Canada, being first reported in eastern Canada in the 1930s and western Canada in the 1970s. However, historical records are scarce and its emergence is poorly understood. The establishment of this parasite in Canada provides an interesting opportunity to explore the use of population genetic approaches to help elucidate the invasion history of a relatively recently established helminth parasite. In this study, we compare the genetic diversity and population structure of a number of D. dendriticum populations from western and eastern Canada, and compare these with much longer established European populations. Two independent genetic marker systems were used; a microsatellite marker panel and a cytochrome c oxidase 1 (cox1) mitochondrial (mt)DNA sequence marker. We found distinct differences in both genetic diversity and population structure of the different Canadian populations that provide insights into their invasion histories compared with the European populations. Two populations from British Columbia, Canada - Salt Spring and Vancouver Islands - are of low diversity, show evidence of a population bottleneck and are closely related to each other, suggesting a shared recent history of establishment. These west coast populations are otherwise most closely related to those from eastern Canada and western Europe, and in contrast are genetically divergent from those in Cypress Hills, Alberta, Canada. Although the Alberta parasite population is the most recently reported in Canada, being first identified there in the early 1990s, it was the most genetically diverse of those examined and showed a strong pattern of admixture of genotypes present in western and eastern Europe. Overall, our results are consistent with a model in which western Europe is likely the source of flukes on the east coast of Canada, which were then subsequently translocated to the west coast of Canada. The most recently reported D. dendriticum population in Canada appears to have a different history and likely has multiple origins.

Life Cycle, Host Utilization, and Ecological Fitting for Invasive Lancet Liver Fluke, Dicrocoelium dendriticum, Emerging in Southern Alberta, Canada.

The expansion of parasite distributions outside of their native host and geographical ranges has occurred repeatedly over evolutionary time. Contemporary examples include emerging infectious diseases (EIDs), many of which pose threats to human, domestic animal, and wildlife populations. Theory predicts that parasites with complex life cycles will be rare as EIDs due to constraints imposed by host specialization at each life-cycle stage. In contrast to predictions of this theory, we report 2 new intermediate hosts in the 3-host life cycle of the liver fluke Dicrocoelium dendriticum in Cypress Hills Provincial Park, Alberta, Canada. Results of sequence analysis of the cytochrome oxidase 1 (cox1) mitochondrial gene identified the terrestrial snail Oreohelix subrudis and the ant Formica aserva as first and second intermediate hosts, respectively, in the region. Neither of these intermediate hosts, nor their suite of domestic and wild mammalian grazers used in the life cycle, occurs within the native range of D. dendriticum in Europe. Our results from host surveys show that the prevalence of D. dendriticum in samples of O. subrudis varied between 4% and 10%, whereas mean metacercariae intensity in F. aserva varied between 33 and 41 (n = 163, mean ± SD = 38 ± 35). These results are the first to describe the complete life cycle of emerging lancet fluke in western North America. The process of multi-level ecological fitting, in which the lancet fluke possesses pre-existing traits to utilize host resources, rather than host species, at each life-cycle stage provides a mechanism for the establishment of this complex life cycle in a novel habitat and in novel hosts.

Parasites and a host's sense of smell: reduced chemosensory performance of fathead minnows (Pimephales promelas) infected with a monogenean parasite.

Parasites residing within the central nervous system of their hosts have the potential to reduce various components of host performance, but such effects are rarely evaluated. We assessed the olfactory acuity of fathead minnows (Pimephales promelas) infected experimentally with the monogenean Dactylogyrus olfactorius, the adults of which live within the host's olfactory chambers. Olfactory acuity was compared between infected and uninfected hosts by assessing electro-olfactography (EOG) neural responses to chemical stimuli that indicate the presence of food (L-alanine) or the presence of conspecifics (taurocholic acid). We also compared differences in gross morphology of the olfactory epithelium in infected and uninfected minnows. Differences in EOG responses between infected and uninfected minnows to both cue types were non-significant at 30 days post-exposure. By days 60 and 90, coincident with a two times increase in parasite intensity in the olfactory chambers, the EOG responses of infected minnows were 70-90% lower than controls. When infected fish were treated with a parasiticide (Prazipro), olfactory acuity returned to control levels by day 7 post-treatment. The observed reduction in olfactory acuity is best explained by the reduced density of cilia covering the olfactory chambers of infected fish, or by the concomitant increase in the density of mucous cells that cover the olfactory chambers. These morphological changes are likely due to the direct effects of attachment and feeding by individual worms or by indirect effects associated with host responses. Our results show that infection of a commonly occurring monogenean in fathead minnows reduces olfactory acuity. Parasite-induced interference with olfactory performance may reduce a fish's ability to detect, or respond to, chemical cues originating from food, predators, competitors or mates.

Dactylogyrus olfactorius n. sp. (Monogenea, Dactylogyridae) from the olfactory chamber of the fathead minnow, Pimephales promelas Rafinesque (Cyprinidae).

Dactylogyrus olfactorius n. sp. (Monogenea) is described from the olfactory chamber of the fathead minnow Pimephales promelas Rafinesque in Alberta, Canada. The new species resembles Dactylogyrus bychowskyi Mizelle, 1937, D. bifurcatus Mizelle, 1937 and D. simplexus Mizelle, 1937, all parasites of Pimephales spp. in North America, in overall size and shape of the anchors and hooks, and in having a male copulatory complex with a tapered tubular penis and bifurcate accessory piece. Diagnostically, D. olfactorius n. sp. has relatively small anchors, hooks of anchor length, and a thin, long dorsal bar and no apparent ventral bar nor 4A hooks. Scanning electron microscopy revealed the body tegument of D. olfactorius n. sp. to be microvillous and in shallow annular folds, while that of D. bifurcatus, occurring on host gills of the same fish, was avillous and in gill-like folds dorsally and ventrally. Partial 28S rDNA sequences revealed significant differences between the two species, supporting establishment of D. olfactorius n. sp. and dispelling the possibility of ecophenotypic effects of site of attachment on morphology.

Characterization of nine microsatellite loci for Dicrocoelium dendriticum, an emerging liver fluke of ungulates in North America, and their use to detect clonemates and random mating.

This study characterizes polymorphic microsatellite loci from adults of the liver fluke Dicrocoelium dendriticum sampled from a population of sympatric beef cattle and wapiti in a region of emergence in southern Alberta, Canada. We also scrutinized the markers to validate their use in studying the population genetics of this complex life cycle parasite. Among the nine loci described, four deviated significantly from Hardy Weinberg Equilibrium (HWE) due to technical artefacts. The remaining five loci were in HWE. These five provided sufficient resolution to identify clonemates produced from the obligate asexual reproduction phase of the life cycle in snails and to assess the impact of non-random transmission of clonemates on measures of FIS, FST and genotypic disequilibrium. Excluding clonemates, we show that the sub-population of worms was in HWE, that average FIS within hosts was 0.003 (p=0.4922) and that there was no population genetic structure among hosts FST=0.001 (p=0.3243). These markers will be useful for studies of Dicrocoelium dendriticum ecology, transmission, and evolution.

Spatial Distribution of Gyrodactylus salmonis (Monogenea) on the Body of Captive Fingerling Oncorhynchus mykiss, Including Attachment Within the Olfactory Chamber.

Gyrodactylus salmonis is a common ectoparasite on the fins and body of North American salmonids in fresh water. In this study, the spatial distribution of G. salmonis on 60 captive hatchery-reared rainbow trout, Oncorhynchus mykiss , is reported. The highest parasite densities occurred on 5 × 5-mm(2) sections of the dorsal fin followed by the trunk, other fins, and the olfactory chamber, with the lowest densities on the head. The finding of infections within the olfactory chamber of 93% of the fish was unexpected. One possibility is that such infections represented spillover from high-density infrapopulations that occur on the skin and fins. However, this possibility is unlikely, because worm densities at various sites along the body surface of infected fish did not correlate with densities within the olfactory chamber. The parasite conceivably enters the chamber either via water incurrent or by crawling in from the head and subsequently remaining at this site to feed and reproduce. Results from scanning electron microscopy are consistent with physical modification to the olfactory epithelium associated with the attachment/reattachment of the opisthaptor and epithelial grazing.

Where's the risk? Landscape epidemiology of gastrointestinal parasitism in Alberta beef cattle.

BACKGROUND: Gastrointenstinal nematodes (GIN) present a serious challenge to the health and productivity of grazing stock around the globe. However, the epidemiology of GIN transmission remains poorly understood in northern climates. Combining use of serological diagnostics, GIS mapping technology, and geospatial statistics, we evaluated ecological covariates of spatial and temporal variability in GIN transmission among bovine calves pastured in Alberta, Canada. METHODS: Sera were collected from 1000 beef calves across Alberta, Canada over three consecutive years (2008-2010) and analyzed for presence of anti-GIN antibodies using the SVANOVIR Ostertagia osteragi-Ab ELISA kit. Using a GIS and Bayesian multivariate spatial statistics, we evaluated the degree to which variation in specific environmental covariates (e.g. moisture, humidity, temperature) was associated with variation in spatial and temporal heterogeneity in exposure to GIN (Nematodirus and other trichostrongyles, primarily Ostertagia and Cooperia). RESULTS: Variation in growing degree days above a base temperature of 5 °C, humidity, air temperature, and accumulated precipitation were found to be significant predictors of broad-scale spatial and temporal variation in serum antibody concentrations. Risk model projections identified that while transmission in cattle from southeastern and northwestern Alberta was relatively low in all years, rate of GIN transmission was generally higher in the central region of Alberta. CONCLUSIONS: The spatial variability in risk is attributed to higher average humidity, precipitation and moderate temperatures in the central region of Alberta in comparison with the hot, dry southeastern corner of the province and the cool, dry northwestern corner. Although more targeted sampling is needed to improve model accuracy, our projections represent an important step towards tying treatment recommendations to actual risk of infection.

Comparative recruitment, morphology and reproduction of a generalist trematode, Dicrocoelium dendriticum, in three species of host.

Epidemiological rate parameters of host generalist parasites are difficult to estimate, especially in cases where variation in parasite performance can be attributed to host species. Such cases are likely common for generalist parasites of sympatric grazing mammals. In this study, we combined data from experimental exposures in cattle and sheep and natural infections in elk to compare the recruitment, morphology and reproduction of adult Dicrocoelium dendriticum, a generalist trematode that has emerged in sympatric grazing hosts in Cypress Hills Provincial Park, Alberta. Overall, there were no significant differences in the recruitment of metacercariae and in the pre-patency period of adults in experimentally exposed cattle and sheep. All flukes reached reproductive maturity and the degree of reproductive inequality between individual flukes within each infrapopulation was moderate and approximately equal among the three host species. Neither fluke size nor per capita fecundity was constrained by density dependence. Thus, fitness parameters associated with growth and reproduction were approximately equivalent among at least three species of definitive host, two of which are sympatric on pastures in this Park. The generalist life-history strategy of this trematode, which is known to extend to other stages of its life cycle, has likely contributed to its invasion history outside its native range in Europe.

Fluke abundance versus host age for an invasive trematode (Dicrocoelium dendriticum) of sympatric elk and beef cattle in southeastern Alberta, Canada.

Epidemiological parameters such as transmission rate, rate of parasite-induced host mortality, and rate of development of host defenses can be assessed indirectly by characterizing the manner in which parasite burdens change with host age. For parasites that are host generalists, estimates of these important parameters may be host-species dependent. In a cross-sectional study, we determined age-abundance profiles of infection in samples of sympatric free-ranging elk and domestic cattle infected with the lancet liver fluke, Dicrocoelium dendriticum. This parasite was introduced into Cypress Hills Provincial Park in southeastern Alberta, Canada in the mid 1990s, and now occurs in 60-90% of co-grazing elk and beef cattle examined at necropsy. The livers of 173 elk were made available by hunters during the 1997-2011 hunting seasons and livers from 35 cattle were purchased from ranchers. In elk, median worm abundance peaked in 6-24 month-olds (median = 72, range = 0-1006) then significantly declined to <10 worms/host in 10-16 year olds. The decline in fluke burden with age is not consistent with an age-related decline in exposure to metacercariae in intermediate hosts and high rates of fluke-induced host mortality are unlikely. Rather, the pattern of peak fluke burdens in elk calves and juveniles, followed by a decline in older animals is consistent with the development of a protective immune response in older hosts. There was no pattern of worm accumulation or decline in sympatric cattle, although statistical power to detect a significant effect was low. These results highlight the complexity and context-dependent nature of epidemiological processes in multi-host systems.

Annual variation in serum antibody concentrations against gastrointestinal nematodes in beef calves from semi-arid rangelands of western Canada.

Sera from calves originating on a 17,000 ha ranch in southeastern Alberta, were collected over three consecutive years (2008-2010) at weaning and analysed for the presence of anti-Ostertagia antibodies using the SVANOVIR(®)Ostertagia ostertagi-Ab ELISA kit. Faecal samples taken at the same time were analysed for presence of nematode eggs using the modified Wisconsin faecal flotation technique. Mean optical density ratio (ODR) values for all calves were similar in 2008 and 2010 while the 2009 values were significantly higher. Similar cumulative frequency distribution profiles were found for 2008 and 2010 while the 2009 profile had an increased number of values in the higher bin categories. This pattern indicates higher nematode transmission on pasture in 2009. There was no significant difference in ODR values between egg positive and egg-negative calves and no significant correlation was detected between individual ODR values and faecal egg output. Significantly higher ODR values in 2009 appear contradictory to general assumptions about the ecological requirements of larval development, as 2009 was a relatively dry year. However, examination of weather data identified 5-6 rainfall events that may have resulted in episodic enhanced survival of nematode larvae.

Shoaling as an antiparasite defence in minnows (Pimephales promelas) exposed to trematode cercariae.

1. Individuals that live in groups benefit from increased foraging success and decreased predation. Protection from some types of parasites may provide an additional benefit of group-living. For fish, the extent to which shoaling can reduce an individual's risk of exposure to the infective stages of parasites is unknown. 2. We tested for antiparasite benefits of shoaling in fathead minnows exposed to larvae (cercariae) of two of their most common species of trematode, Ornithodiplostomum ptychocheilus and Posthodiplostomum minimum. As developing stages (metacercariae) of these trematodes cause reductions in minnow activity, growth and survival, natural selection should favour the evolution of cercariae-avoidance behaviours. 3. We evaluated shoal dimensions in groups of minnows exposed to O. ptychocheilus and to other chemical/physical stimuli within aquaria. To compare risk of exposure in shoaling vs. non-shoaling fish, we confined groups of minnows into mesh cages in outdoor mesocosms, exposed them to cercariae, then compared mean worm numbers in grouped vs. solitary fish. Lastly, we tested whether fish located within the centre of an artificial shoal reduced their risk of cercariae exposure compared with those along peripheral edges. 4. Minnows distinguished infective cercariae from other potential aquatic threats and responded with activity that reduced the 2-dimensional area of their shoals 15-fold compared with water-only controls. Fish confined within artificial shoals had 3-fold fewer worms than single fish and minnows located within the centre of artificial shoals had significantly fewer worms than those without peripheral minnows. 5. These results show that shoaling reduces a minnows' risk of exposure to cercariae, either directly via detection of cercariae in the water column followed by behavioural avoidance or indirectly via behaviour-mediated differences in exposure between shoaling vs. non-shoaling fish.