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Ecology of the brain trematode Euhaplorchis californiensis and its host, the California Killifish (Fundulus parvipinnis)

We describe the distribution and abundance of the brain-encysting trematode Euhaplorchis californiensis and its second intermediate host, the California killifish ( Fundulus parvipinnis ), in 3 estuaries in southern California and Baja California. We quantified the density of fish and metacercariae at 13–14 sites per estuary and dissected 375 killifish. Density (numbers and biomass) was examined at 3 spatial scales, i.e., small replicate sites, habitats, and entire estuaries. At those same scales, factors that might influence metacercaria prevalence, abundance, and aggregation in host individuals and populations were also examined. Metacercaria prevalence was 94–100% among the estuaries. Most fish were infected with 100s to 1,000s of E. californiensis metacercariae, with mean abundance generally increasing with host size. Although body condition of fish did not vary among sites or estuaries, the abundance of metacercariae varied significantly among sites, habitats, estuaries, and substantially with host size and gender. Metacercariae were modestly aggregated in killifish ( k > 1), with aggregation decreasing in larger hosts. Across the 3 estuaries, the total populations of killifish ranged from 9,000–12,000 individuals/ha and from 7–43 kg/ha. The component populations of E. californiensis metacercariae ranged from 78–200 million individuals/ha and from 0.1–0.3 kg/ha. Biomass of E. californiensis metacercariae constituted 0.5–1.7% of the killifish biomass in the estuaries. Our findings, in conjunction with previously documented effects of E. californiensis , suggest a strong influence of this parasite on the size, distribution, biomass, and abundance of its killifish host.

Journal of Parasitology

Thermal constraints to the sporogonic development and altitudinal distribution of avian malaria Plasmodium relictum in Hawai'i

More than half of the Hawaiian honeycreepers (Drepanidinae) known from historical records are now extinct. Introduced mosquito-borne disease, in particular the avian malaria Plasmodium relictum , has been incriminated as a leading cause of extinction during the 20th century and a major limiting factor in the recovery of remaining species populations. Today, most native Hawaiian bird species reach their highest densities and diversity in high elevation (>1,800 m above sea level) forests. We determined the thermal requirements for sporogonic development of P. relictum in the natural vector, Culex quinquefasciatus , and assessed the current distribution of native bird species in light of this information. Sporogonic development was completed at constant laboratory and mean field temperatures between 30 and 17 C, but development, prevalence, and intensity decreased significantly below 21 C. Using a degree-day (DD) model, we estimated a minimum threshold temperature of 12.97 C and a thermal requirement of 86.2 DD as necessary to complete development. Predicted (adiabatic lapse-rate) and observed summer threshold isotherm (13 C) correspond to the elevation of high forest refuges on the islands of Maui and Hawai'i. Our data support the hypothesis that avian malaria currently restricts the altitudinal distribution of Hawaiian honeycreeper populations and provide an ecological explanation for the absence of disease at high elevation.

Hawaii

Release of infectious cells from epidermal ulcers in Ichthyophonus sp.–infected Pacific Herring (Clupea pallasii): Evidence for multiple mechanisms of transmission

A common clinical sign of ichthyophoniasis in herring and trout is “sandpaper” skin, a roughening of the epidermis characterized by the appearance of small papules, followed by ulceration and sloughing of the epithelium; early investigators hypothesized that these ulcers might be a means of transmitting the parasite, Ichthyophonus sp ., without the necessity of ingesting an infected host. We examined the cells associated with the epidermal lesions and confirmed that they were viable Ichthyophonus sp. cells that were readily released from the skin into the mucous layer and ultimately into the aquatic environment. The released cells were infectious when injected into the body cavity of specific-pathogen-free herring. Our hypothesis is that different mechanisms of transmission occur in carnivorous and planktivorous hosts: Planktonic feeders become infected by ingestion of ulcer-derived cells, while carnivores become infected by ingestion of whole infected fish.

Journal of Parasitology

Infection by Haemoproteus parasites in four species of frigatebirds and the description of a new species of Haemoproteus (Haemosporida: Haemoproteidae)

Among seabirds, the fregatids stand out with a high prevalence of blood parasites. Four of 5 species in this family have been found to be infected with Haemoproteus ; however, complete species descriptions with molecular phylogeny are lacking. Seventy-five samples from 4 species of frigatebirds, i.e., Fregata andrewsi , Fregata minor , Fregata magnificens , and Fregata aquila , were screened for infections caused by species of Haemoproteus . Four different parasite haplotypes were found infecting frigatebirds based on the sequencing of a fragment of the cytochrome b gene. Two haplotypes belong to the subgenus Parahaemoproteus , and the other 2 correspond to haplotypes within the subgenus Haemoproteus . The more prevalent and cosmopolitan Parahaemoproteus haplotype (FregPHae1) was phylogenetically grouped with other Haemoproteus parasites infecting non-passerine birds, but it could not be detected from the single sample from F. aquila . The other Parahaemoproteus haplotype (FregPHae2) was not phylogenetically clustered with parasites infecting non-passerine birds, and it was sequenced from a single (1 each) F. andrewsi and F. minor . Blood smears from F. andrewsi infected only by FregPHae1 haplotype showed sufficient gametocytes to allow description of a new species, Haemoproteus valkiūnasi sp. nov. In contrast to Haemoproteus iwa , the only previously known blood parasite infecting frigatebirds and described from F. minor from Galapagos Islands, parasites from F. andrewsi (1) are shorter with no contact of gametocyte with host cell membrane, (2) have fewer pigment granules, and (3) have wider microgametocytes, with a smaller host nuclear displacement. In contrast, patent single infections corresponding to the cosmopolitan haplotype of the subgenus Haemoproteus (FregHae1) were also found in samples from 1 F. andrewsi , 1 F. minor , and 1 F. aquila . In all these cases, the number of microgametocytes was very low, resembling H. iwa , which lacks microgametocytes in the original description. Macrogametocytes of haplotype FregHae1 in F. andrewsi differ significantly from all the characteristics measured from H. valkiūnasi . In addition, it also differs from all characteristics of H. iwa despite being genetically identical in the analyzed fragment.

Journal of Parasitology

Parasites of fishes in the Colorado River and selected tributaries in Grand Canyon, Arizona.

As part of the endangered humpback chub (HBC; Gila cypha) Adaptive Management Program, a parasite survey was conducted from 28 June to 17 July 2006 in 8 tributaries and 7 adjacent sections of the main stem of the Colorado River, U.S.A. In total, 717 fish were caught, including 24 HBC. Field necropsies yielded 19 parasite species, 5 of which (Achtheres sp., Kathlaniidae gen. sp., Caryophyllaidae gen. sp., Myxidium sp., and Octomacrum sp.) are new records for Grand Canyon, Arizona, U.S.A. Spearman's correlation coefficient analyses showed no correlations between parasite burden and fork length for various combinations of fish and parasite species. Regression analyses suggest that no parasite species had a strong effect on fish length. The most diverse parasite community (n=14) was at river kilometer (Rkm) 230, near the confluence of Kanab Creek. The most diverse parasite infracommunity (n=12) was found in the non-native channel catfish (CCF; Ictaluris punctatus). Overall parasite prevalence was highest in CCF (85%) followed by that in HBC (58%). The parasite fauna of humpback chub was mainly composed of Bothriocephalus acheilognathi and Ornithodiplostomum sp. metacercariae.

Arizona

Diagnostic methodology is critical for accurately determining the prevalence of ichthyophonus infections in wild fish populations

Several different techniques have been employed to detect and identify Ichthyophonus spp. in infected fish hosts; these include macroscopic observation, microscopic examination of tissue squashes, histological evaluation, in vitro culture, and molecular techniques. Examination of the peer-reviewed literature revealed that when more than 1 diagnostic method is used, they often result in significantly different results; for example, when in vitro culture was used to identify infected trout in an experimentally exposed population, 98.7% of infected trout were detected, but when standard histology was used to confirm known infected tissues from wild salmon, it detected ~50% of low-intensity infections and ~85% of high-intensity infections. Other studies on different species reported similar differences. When we examined a possible mechanism to explain the disparity between different diagnostic techniques, we observed non-random distribution of the parasite in 3-dimensionally visualized tissue sections from infected hosts, thus providing a possible explanation for the different sensitivities of commonly used diagnostic techniques. Based on experimental evidence and a review of the peer-reviewed literature, we have concluded that in vitro culture is currently the most accurate diagnostic technique for determining infection prevalence of Ichthyophonus, particularly when the exposure history of the population is not known.

Journal of Parasitology

The role of spatial and temporal heterogeneity and competition in structuring trematode communities in the great pond snail, Lymnaea stagnalis (L.)

We assessed how spatial and temporal heterogeneity and competition structure larval trematode communities in the pulmonate snail Lymnaea stagnalis . To postulate a dominance hierarchy, mark-release-recapture was used to monitor replacements of trematode species within snails over time. In addition, we sampled the trematode community in snails in different ponds in 3 consecutive years. A total of 7,623 snails (10,382 capture events) was sampled in 7 fishponds in the Jindřichův Hradec and Třeboň areas in South Bohemia (Czech Republic) from August 2006 to October 2008. Overall, 39% of snails were infected by a community of 14 trematode species; 7% of snails were infected with more than 1 trematode species (constituting 16 double- and 4 triple-species combinations). Results of the null-model analyses suggested that spatial heterogeneity in recruitment among ponds isolated trematode species from each other, whereas seasonal pulses in recruitment increased species interactions in some ponds. Competitive exclusion among trematodes led to a rarity of multiple infections compared to null-model expectations. Competitive relationships among trematode species were hypothesized as a dominance hierarchy based on direct evidence of replacement and invasion and on indirect evidence. Seven top dominant species with putatively similar competitive abilities (6 rediae and 1 sporocyst species) reduced the prevalence of the other trematode species developing in sporocysts only.

Journal of Parasitology

New species of Parapharyngodon (Nematoda: Pharyngodonidae) in Rhinella marina (Anura: Bufonidae) from Grenada, West Indies

Parapharyngodon grenadaensis n. sp. (Nematoda: Pharyngodonidae) from the large intestine of the cane toad, Rhinella marina, is described and illustrated. Parapharyngodon grenadaensis n. sp. is the 48th species assigned to the genus and the 16th species from the Neotropical region. It differs from other species in the genus by possessing 4 pairs of caudal papillae, an echinate anterior cloacal lip, and a blunt spicule of 67–104 μm. This is only the second report of R. marina harboring a species of Parapharyngodon.

Journal of Parasitology

Evidence for an amoeba-like infectious stage of ichthyophonus sp. and description of a circulating blood stage: a probable mechanism for dispersal within the fish host

Small amoeboid cells, believed to be the infectious stage of Ichthyophonus sp., were observed in the bolus (stomach contents) and tunica propria (stomach wall) of Pacific staghorn sculpins and rainbow trout shortly after they ingested Ichthyophonus sp.–infected tissues. By 24–48 hr post-exposure (PE) the parasite morphed from the classically reported multinucleate thick walled schizonts to 2 distinct cell types, i.e., a larger multinucleate amoeboid cell surrounded by a narrow translucent zone and a smaller spherical cell surrounded by a “halo” and resembling a small schizont. Both cell types also appeared in the tunica propria, indicating that they had recently penetrated the columnar epithelium of the stomach. No Ichthyophonus sp. pseudo-hyphae (“germination tubes”) were observed in the bolus or penetrating the stomach wall. Simultaneously, Ichthyophonus sp. was isolated in vitro from aortic blood, which was consistently positive from 6 to 144 hr PE, then only intermittently for the next 4 wk. Small PAS-positive cells observed in blood cultures grew into colonies consisting of non-septate tubules (pseudo-hyphae) terminating in multinucleated knob-like apices similar to those seen in organ explant cultures. Organ explants were culture positive every day; however, typical Ichthyophonus sp. schizonts were not observed histologically until 20–25 days PE. From 20 to 60 days PE, schizont diameter increased from ≤25 μm to ≥82 μm. Based on the data presented herein, we are confident that we have resolved the life cycle of Ichthyophonus sp. within the piscivorous host.

Journal of Parasitology

Occurrence of Ergasilus megaceros Wilson, 1916, in the sea lamprey and other fishes from North America

Ergasilus megaceros (Copepoda: Ergasilidae) was recovered from the nasal fossae (lamellae) of the olfactory sac in 1 (1.8%) of 56 sea lampreys, Petromyzon marinus Linne, 1758, collected in May 2002 from the Cheboygan River, Michigan. Although the sea lamprey is a new host record for E. megaceros , this fish species may not be a preferred host because of its low prevalence. Ergasilus megaceros is the second ergasilid species reported from the sea lamprey in North America. This is the third report of an ergasilid species infecting the nasal fossae of fishes in North America, with E. rhinos being the only other species reported from this site.

Journal of Parasitology

The severity of Whirling disease among wild trout corresponds to the differences in genetic composition of tubifex tubifex populations in Central Colorado

We analyzed the geographic distribution of Tubifex tubifex from various river drainages in central Colorado by genetic screening with specific mitochondrial 16S ribosomal DNA (mt 16S rDNA) markers. Four distinct mt 16S rDNA lineages are evident. The sites varied with respect to land- and water-use practices. All sites represented habitats presumed capable of supporting oligochaetes. At the locations where whirling disease has had the greatest impact on resident rainbow trout, T. tubifex, representing lineages I and III (genotypes known to be susceptible to Mxyobolus cerebralis), were most commonly found. In contrast, at sites less affected by whirling disease, T. tubifex of lineages V and VI that are more resistant to M. cerebralis infections were more abundant. The predominance of resistant T. tubifex worms (lineages V and VI) at low-impact sites supports the conclusion that when these genotypes are in greater abundance, the potential for more severe effects of whirling disease on wild rainbow trout populations may be diminished. ?? American Society of Parasitologists 2005.

Journal of Parasitology

Differential propagation of the metazoan parasite Myxobolus cerebralis by Limnodrilus hoffmeisteri, Ilyodrilus templetoni, and genetically distinct strains of Tubifex tubifex

Whirling disease, caused by the parasite Myxobolus cerebralis, has infected rainbow trout (Oncorhynchus mykiss) and other salmonid fish in the western United States, often with devastating results to native populations but without a discernible spatial pattern. The parasite develops in a complex 2-host system in which the aquatic oligochaete Tubifex tubifex is an obligate host. Because substantial differences in whirling disease severity in different areas of North America did not seem explainable by environmental factors or features of the parasite or its fish host, we sought to determine whether ecological or genetic variation within oligochaete host populations may be responsible. We found large differences in compatibility between the parasite and various laboratory strains of T. tubifex that were established from geographic regions with different whirling disease histories. Moreover, 2 closely related species of tubificids, Limnodrilus hqffmeisteri and Ilyodrilus templetoni, which occur naturally in mixed species assemblages with T. tubifex, were incompatible with M. cerebralis. Virulence of the parasite was directly correlated with the numbers of triactinomyxon spores that developed within each strain of T. tubifex. Thus, the level of virulence was directly related to the compatibility between the host strain and the parasite. Genetic analyses revealed relationships that were in agreement with the level of parasite production. Differences in compatibilities between oligochaetes and M. cerebralis may contribute to the spatial variance in the severity of the disease among salmonid populations. ?? American Society of Parasitologists 2004.

Journal of Parasitology

Comparative susceptibility of introduced forest-dwelling mosquitoes in Hawai'i to avian malaria, Plasmodium relictum

To identify potential vectors of avian malaria in Hawaiian native forests, the innate susceptibility of Aedes albopictus, Wyeomyia mitchellii, and Culex quinquefasciatus from 3 geographical sites along an altitudinal gradient was evaluated using local isolates of Plasmodium relictum. Mosquitoes were dissected 5-8 and 9-13 days postinfective blood meal and microscopically examined for oocysts and salivary-gland sporozoites. Sporogony was completed in all 3 species, but prevalence between species varied significantly. Oocysts were detected in 1-2% and sporozoites in 1-7% of Aedes albopictus that fed on infected ducklings. Wyeomyia mitchellii was slightly more susceptible, with 7-19% and 7% infected with oocysts and sporozoites, respectively. In both species, the median oocyst number was 5 or below. This is only the second Wyeomyia species reported to support development of a malarial parasite. Conversely, Culex quinquefasciatus from all 3 sites proved very susceptible. Prevalence of oocysts and sporozoites consistently exceeded 70%, regardless of gametocytemia or origin of the P. relictum isolate. In trials for which a maximum 200 oocysts were recorded, the median number of oocysts ranged from 144 to 200. It was concluded that Culex quinquefasciatus is the primary vector of avian malaria in Hawai'i. ?? American Society of Parasitologists 2005.

Journal of Parasitology

An efficient strategy to estimate intensity and prevalence: Sampling metacercariae in fishes

Accurate estimates of population-level parameters of parasites, such as prevalence and mean intensity, require large sample sizes. The processing of such samples becomes an overwhelming task when parasites are abundant, as with trematode metacercariae in fishes. In the present study, a subsampling method reduced processing time while maintaining an accurate estimation of metacercariae prevalence and intensity across 3 trematode species and 2 fish species. By double sampling, we generated regression models to predict total intensity from a combination of subsamples. The key to this approach lies in choosing the best strategy from a large number of potential subsampling routines. We selected the most efficient routine by weighing the costs and benefits of each. This approach, however, could not provide an estimate of parasite abundance when no parasites occurred in the initial subsample. To estimate prevalence accurately, our subsampling algorithm prescribed an additional sampling sequence using a new, optimal regression model. In addition, we optimized the technique to measure three parasite species infecting a single host simultaneously. This efficient subsampling procedure decreased the overall processing time per host by up to 91% while obtaining accurate ( R 2 > 0.8) estimates for both prevalence and intensity.

Journal of Parasitology

Molecular phylogeny of Babesia poelea from brown boobies ( Sula leucogaster ) from Johnston Atoll, Central Pacific

The phylogenetic relationship of avian Babesia with other piroplasms remains unclear, mainly because of a lack of objective criteria such as molecular phylogenetics. In this study, our objective was to sequence the entire 18S, ITS-1, 5.8S, and ITS-2 regions of the rRNA gene and partial ß-tubulin gene of B. poelea , first described from brown boobies ( Sula leucogaster ) from the central Pacific, and compare them to those of other piroplasms. Phylogenetic analyses of the entire 18S rRNA gene sequence revealed that B. poelea belonged to the clade of piroplasms previously detected in humans, domestic dogs, and wild ungulates in the western United States. The entire ITS-1, 5.8S, ITS-2, and partial ß-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the ß-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm.

Johnston Atoll National Wildlife Refuge

Evidence of experimental postcyclic transmission of Bothriocephalus acheilognathi in bonytail chub (Gila elegans)

We examined the role that predation of infected conspecific fish and postcyclic transmission might play in the life cycle of the Asian fish tapeworm, Bothriocephalus acheilognathi (Cestoda: Pseudophyllidea) Yamaguti, 1934. Young-of-the-year (YOY) bonytail chub ( Gila elegans ) were exposed to copepods infected with B. acheilognathi and subsequently fed to subadult bonytail chub. Within 1 wk after consumption of the YOY chub, subadults were necropsied and found infected with gravid and nongravid tapeworms. This study provides evidence that postcyclic transfer of B. acheilognathi can occur. Postcyclic transmission may be an important life history trait of B. acheilognathi that merits consideration when studying the impact and distribution of this invasive and potentially pathogenic tapeworm.

New Mexico

Ectoparasites in an urban population of big brown bats (Eptesicus fuscus) in Colorado

Ectoparasites of an urban population of big brown bats (Eptesicus fuscus) in Fort Collins, Colorado, were investigated during summers 2002, 2003, and 2004. Eleven species of ectoparasites were found (the macronyssid mite Steatonyssus occidentalis, the wing mite Spinturnix bakeri, the myobiid mites Acanthophthirius caudata and Pteracarus aculeus, the chirodiscid mite Alabidocarpus eptesicus, the demodicid mite Demodex sp., the chigger Leptotrombidium myotis, the soft tick Carios kelleyi, the batfly Basilia forcipata, the batbug Cimex pilosellus, and the flea Myodopsylla borealis). Five species were analyzed by prevalence and intensity (C. pilosellus, M. borealis, L. myotis, S. bakeri, and S. occidentalis) based on 2,161 counts of 1,702 marked individual bats over the 3 summer study periods. We investigated 4 factors potentially influencing prevalence and intensity: age class of the host, reproductive status of adult female hosts, roosts in which the hosts were found, and abiotic conditions during the year sampled. The macronyssid mite, S. occidentalis, was the most prevalent and abundant ectoparasite. Adult big brown bats had more ectoparasites than volant juveniles for most of the species analyzed. In a sample of known age bats at 1 large colony, bats of 4 yr of age or greater had higher ectoparasite loads of S. occidentalis and S. bakeri when compared with younger bats. Lactating female bats had the highest prevalence and intensities of most ectoparasites. Annual differences in ectoparasite prevalence and intensity were related to temperature and humidity, which can affect the nidicolous species of ectoparasites. Residents of 2 buildings sprayed insecticides in response to Cimex sp., and this appeared to reduce ectoparasitism of S. occidentalis and C. pilosellus present at these buildings. Intensity of S. occidentalis had no influence on annual survival of big brown bats. ?? American Society of Parasitologists 2007.

Journal of Parasitology

Blood parasites of birds of the District of Columbia and Patuxent Research Refuge vicinity

Recently a number of surveys of blood parasites of birds have been made in different parts of this country by different workers, the general purpose of which was to further the knowledge of the distribution and pathogenicity of these protozoa. This paper treats the distribution of parasites found in sampling miscellaneous birds of the District of Columbia and vicinity and contains a preliminary report on the more important phase of the problem of pathogenicity.

Maryland