Using transmission electron microscopy, spermiogenesis and the spermatozoon ultrastructural organization are described in Ligula intestinalis (Linnaeus, 1758) (Diphyllobothriidea), a parasite of the great crested grebe Podiceps cristatus (Linnaeus, 1758). Spermiogenesis starts with the differentiation zone of 2 striated rootlets, 2 centrioles giving rise to 2 flagella, and an intercentriolar body. The latter is composed of 5 electron-dense layers separating 4 electron-lucent layers. In the early stages of spermiogenesis, an electron-dense material is present in the apical region of the differentiation zone. Later, the flagella undergo a rotation and fuse with the cytoplasmic extension in a proximo-distal process. The spermatozoon contains 2 axonemes with a 9 “1” trepaxonematan pattern, the nucleus, the cortical microtubules, and an electron-dense zone. The spermatozoon anterior extremity in L. intestinalis is characterized by the absence of crested bodies and a ring of electron-dense cortical microtubules. Some characters of spermiogenesis and spermatozoon in L. intestinalis confirm the recent splitting of “Pseudophyllidea” into 2 new orders, i.e., Bothriocephalidea and Diphyllobothriidea. The process of spermiogenesis is similar in both orders for the “type I” of spermiogenesis and the presence of electron-dense material. However, the intercentriolar body is clearly more developed in the Diphyllobothriidea than in the Bothriocephalidea. Moreover, these 2 orders seem to differ in the presence or absence of a ring of electron-dense cortical microtubules in the anterior extremity of the spermatozoon.

Parasites are incredibly diverse. An important factor in the evolution of this diversity is the fact that many parasite species are restricted to 1, or just a few, host species. In addition, some parasites exhibit geographic specificity that is nested within their specificity to a particular species of host. The environmental factors that restrict parasites to particular regions within the host's range are poorly understood, and it is often difficult to know whether such patterns of geographic specificity are real, or merely artifacts of uneven host sampling. For over a decade, we sampled communities of ectoparasitic lice (Phthiraptera) from western scrub-jays (Aphelocoma californica) throughout their range in the United States, and found 3 common species of lice. Philopterus crassipes was found throughout the host range, whereas the other 2 species of lice had more restricted distributions. Brueelia deficiens was found only on the woodhouseii host subspecies group, and Myrsidea sp. was found largely on the californica host subspecies group. We suggest that differential tolerance to arid conditions and interspecific competition has led to the restricted geographic distributions of these 2 species of lice.

Given the importance of temperature for the shedding of trematode infective stages (cercariae) from gastropod first intermediate hosts, we investigated the response to temperature increases of trematodes of the intertidal snail Zeacumantus subcarinatus collected from different latitudes (differing in mean annual summer temperature) on the South Island of New Zealand. We investigated whether shedding of cercariae at elevated temperatures differed both between species (interspecific variation) and among populations of each trematode species (intraspecific variation). The shedding of one trematode species, Maritrema novaezealandensis (Microphallidae), appears to decrease with increased temperature and differed among locations. In contrast, the shedding of a second species, Acanthoparyphium sp. (Echinostomatidae), increased at warmer temperatures. In particular, Acanthoparyphium sp.–infected snails from 1 location showed the greatest increase in the shedding of cercariae in response to elevated temperature. Our results demonstrate that different trematode species and populations may be dissimilarly impacted by temperature changes resulting from global warming. In addition, both interspecific and intraspecific variation may result in different impacts of trematodes on ecosystems at different locales.

There are many impressive examples of host manipulation by parasites, but mechanisms underlying these ethological changes, as well as their physiological consequences, are not well characterized. Here, we analyzed part of the cerebral proteome of brine shrimp Artemia infected by manipulative cestodes, using for the first time the ProteinChip Surface-Enhanced Laser Desorption Ionization and Time of Fly Mass Spectrometry (SELDI-TOF MS) system, which has been proposed as an excellent way to analyze the host genome during the host–parasite interaction processes. We found 2 peptides downregulated in individuals infected by the dilepidid, Anomotaenia tringae (4.5 kDa), and by the 2 hymenolepidids, Flamingolepis liguloides and Confluaria podicipina (3.9 kDa), which are potential candidates for involvement with the manipulation process. The identification of 2 head peptides (4.1 and 4.2 kDa) overexpressed in all the categories in brine shrimp living at the surface (both infected individuals and uninfected controls) suggests its association with the different environmental conditions experienced at the water surface. In parallel, brine shrimp infected by C. podicipina showed significant values of triglycerides, potentially augmenting their profitability and attractiveness for the predaceous definitive host (grebes). We discuss our findings in relationship with current ideas on the complexity of parasitically modified organisms.

Philureter trigoniopsis is the only parasite found in the ureters and urinary bladder of Galaxias maculatus in Patagonian Andean lakes. The dynamics of this endoparasitic monogenean were studied in Lake Gutiérrez, a body of water with scarce shoreline vegetation, where the host has an annual cycle of migration to the deep pelagic zone of the lake. To compare variations of the infection related to differences between lakes, G. maculatus specimens were sampled monthly with baited traps from September 1998 to November 1999 in Lake Moreno, which is an oligotrophic body of water with emergent shoreline vegetation and where the fish do not migrate to the deep pelagic areas of the lake. In addition, data for summer infections of P. trigoniopsis from 10 Andean Patagonian lakes that differ in aquatic vegetation, depth, and area were compared. In Lake Moreno prevalence of P. trigoniopsis showed a seasonal pattern, with 1-yr-old fish exhibiting the highest values of prevalence and mean intensity. Negative correlations between water temperature and prevalence and between age of fish and abundance were found. Our results suggest that age of fish may be the main factor structuring the distribution of P. trigoniosis in populations of G. maculatus. At the regional level the relationship between the infection and the characteristics of the lakes was also observed, with prevalence and mean intensity of P. trigoniopsis in G. maculatus higher in large deep lakes without macrophytes.

The objective of this study was to show which species of flies are responsible for human myiasis in the Brazilian state of Goiás and to determine the frequency of cases. Patients at the Clinical Hospital of the Federal University of Goiás (UFG) were examined, and any fly larvae found in their wounds were collected for taxonomic identification. First instar larvae were observed using light microscopy; second and third instars were examined using stereoscopy. The following screwworm flies were observed, in decreasing order of prevalence: Cochliomyia hominivorax, Sarcodexia lambens, Dermatobia hominis, Chrysomya albiceps, Chrysomya megacephala, Lucilia cuprina, and Eristalis tenax. Myiasis was most frequent in the legs in adults, male patients, elderly people, and people of reproductive age. It was lowest in children, females, and patients with neurologic or psychiatric disorders. Frequency was high in patients living in low socioeconomic conditions with poor personal hygiene. Education and sanitation measures are needed to counteract this situation.

During a parasite survey, we collected data on the presence and distribution of feather mites, intestinal parasites, and blood parasites of small ground finches (Geospiza fuliginosa) from 4 islands in the Galapagos. We recorded 4 species of feather mites, with the most common species, Trouessartia geospiza, present on the majority (77% [308/400]) of individuals. Birds with high loads of T. geospiza came from larger islands and had higher body masses. We identified 3 species of intestinal Isospora (Isospora fragmenta, Isospora temeraria, and Isospora exigua) in fecal samples that showed a diurnal pattern of oocyst release. Among samples collected in the afternoon, infection prevalence was 61% (11/18), while only 0.5% (1/192) contained oocysts in the morning. We screened 40 individuals from one island (Isabela) for blood parasites using molecular markers. Although no parasites of Haemoproteus, Leucocytozoon, or Plasmodium were detected, a high proportion of birds (80% [32/40]) had systemic Isospora spp. infections. A high infection prevalence (74% [20/27]), but low infection intensity, was confirmed using optical microscopy. This result could either be due to the detection of a previously unidentified systemic Isospora sp. parasite, or a result of the previously described Isospora spp. parasites causing systemic infections.

Diaphorocleidus orthodusus n. sp. and Diaphorocleidus kabatai (Molnar, Hanek and Fernando, 1974) Jogunoori, Kritsky, and Venkatanarasaiah, 2004 are detailed from Astyanax orthodus and Astyanax aeneus, respectively. Palombitrema heteroancistrium (Price and Bussing, 1968) is described from specimens collected from A. aeneus, and Urocleidoides strombicirrus (Price and Bussing, 1967) is reported for the first time from A. aeneus and Astyanax fasciatus in Panama. Characithecium costaricensis (Price and Bussing, 1967) n. comb. is detailed based on specimens from A. aeneus. Characithecium costaricensis is characterized by having overlapping gonads, a medioventral vaginal aperture, a copulatory complex consisting of single counterclockwise coil of the copulatory organ that is articulated to the accessory piece, and a haptor having 2 pairs of anchors, dorsal and ventral bars, and 14 hooks. Measurements of body size varied substantially among individual worms, both within and across different host species and locations. However, the morphological differences were insufficient to separate species of Characithecium. This result suggests limited parasite speciation across sympatric species of Astyanax in Mexico and Panama.

A variety of demographic, seasonal, and site-specific variables may influence parasitism, but the relative importance of these variables is generally unclear. We measured the relative ability of host characteristics, season, and site to explain louse (Trichodectes octomaculatus) and flea (Orchopeas howardi) infestation across 10 populations of raccoons (Procyon lotor). Lice are highly dependent on specific hosts and are predicted to display a relatively strong relationship with factors intrinsic to the host, when compared to fleas, which can infest multiple species and survive off-host for weeks without feeding. We developed a priori models that represented explicit hypotheses and contrasted their ability to predict infestation patterns. While the abundance of lice was seasonal, models that included solely host age and sex best predicted prevalence and abundance, in part because males were infested with 3 times the number of lice than were females. Conversely, flea prevalence and abundance, which peaks sharply in the spring, was best predicted by season; factors intrinsic to the host were relatively unimportant for predicting abundance. These, and other, recent findings emphasize the need to simultaneously assess the relative importance of multiple ecological variables between parasite species when attempting to describe general trends and constraints of host–parasite associations.

We quantified Neobenedenia melleni from the skin of Caribbean surgeonfishes (Acanthuridae) from June through October 2005 and 2007. Prevalence, or mean intensity of infection, or both, varied significantly among the 3 species, and among sites and between years for the most heavily infected species, blue tang (Acanthurus coeruleus). Among 6 sites sampled, no more than 12% of ocean surgeonfish (Acanthurus bahianus) were infected, compared with 10 to 100% of A. coeruleus. The prevalence of infection among doctorfish (Acanthurus chirurgus), collected at only 1 of the sites, was intermediate between the other 2 species (46%). Mean intensity (range) of infection for the few infected A. bahianus was 1 (1) to 3 (1–8), compared with 1.3 (1–2) to 14.3 (1–59) for A. coeruleus, and 2.5 (1–8) for A. chirurgus. Expected abundance of N. melleni on A. coeruleus from shallow bay sites was greater than for those from non-bay sites. Higher infections on A. coeruleus may be attributable to differences in habitat use, or susceptibility to infection, or both, compared to other species. Among site and between-year differences may be associated with differences in benthic habitat, or water conditions, or both. This system seems ideal for future comparative studies on the relationship between environmental variables and parasites on Caribbean coral reefs.

Haplobothrium globuliforme maintains its position in the proximal mid-gut epithelium of Amia calva with the aid of tentacles, i.e., proboscides, everted from scolices of a primary strobila and craspedote proglottids of a secondary strobila. Weakly developed scolices of the secondary strobila appear to have little holdfast action, but the distinctly craspedote proglottids of these individuals project into the intestinal mucosa, altering the configuration of gut epithelial cells and pushing the tapeworm deeper into mucosal crypts. The basement membrane underlying the epithelium appears to act as a barrier that prevents tapeworms from penetrating into the deeper tissue layers of the lamina propria, muscularis mucosa, or submucosa. Scolex tegument modification occurs at the point of contact with host basement membrane. A mild background infiltrate of lymphocytes and granulocytes was evident adjacent to the scolex and proglottid tegument. There was no evidence of blood vessel proliferation, edema, mast cell degranulation, eosinophilia, or subsequent collagen formation associated with tapeworm activity.

Cryptosporidium parvum, an apicomplexan parasite transmitted via animal fecal wastes, is the causative agent of cryptosporidiosis. Clones were selected from 2 synthetic naïve human single-chain variable fragment (scFv) phagemid libraries that bound to the recombinant P23 protein of C. parvum. Panning the Tomlinson I and J phagemid libraries resulted in 6 distinct clones. Two clones had full-length scFv sequences, while the remaining clones were either truncated or missing a section of the heavy chain. Despite these differences, all clones were able to detect both native C. parvum proteins and recombinant P23. None of the selected clones cross-reacted with Escherichia coli, Streptococcus pyogenes, Listeria monocytogenes, Bacillus cereus, Giardia lamblia (cysts or trophozoites), or with S16, another dominant surface antigen on C. parvum sporozoites. Clones expressed as the scFv-gIIIp fusion construct in soluble form detected C. parvum. Panning from naïve libraries is a useful method for isolation and identification of recombinant antibodies that have the potential for use in pathogen detection and immunotherapy.

Toxoplasma gondii infection in marine mammals is intriguing and indicative of contamination of the ocean environment and coastal waters with oocysts. Toxoplasma gondii infection was detected in captive marine mammals at a sea aquarium in Canada. Antibodies to T. gondii were found in all 7 bottlenose dolphins (Tursiops truncatus) tested. Two of these dolphins, as well as a walrus (Odobenus rosmarus) at the facility, died. Encephalitis and T. gondii tissue cysts were identified in histological sections of the brain of 1 dolphin (dolphin no. 1). Another dolphin (dolphin no. 2) had mild focal encephalitis without visible organisms, but viable T. gondii was isolated by bioassay in mice and cats from its brain and skeletal muscle; this strain was designated TgDoCA1. The PCR-RFLP typing using 11 markers (B1, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) identified a Type II strain. The DNA sequencing of B1 and SAG1 alleles amplified from TgDoCA1 and directly from the brains of dolphin no. 1 and the walrus showed archetypal alleles consistent with infection by a Type II strain. No unique polymorphisms were detected. This is apparently the first report of isolation of T. gondii from a marine mammal in Canada.

Neospora caninum is a well known protozoan parasite of domestic and wild animals. Neospora hughesi is a closely related protozoan with an unknown life cycle, host range, and infection prevalence. Many serologic surveys of N. caninum have been performed without consideration of potential cross-reactions with N. hughesi, which could confound results. The aim of this study was to investigate whether postexposure sera from animals experimentally infected with N. caninum exhibit significant reactivity differences when tested using N. caninum and N. hughesi Immunofluorescent Antibody Tests (IFAT). Pre- and postinfection serum samples from 10 dogs, 20 calves, and 17 cows were tested by dual IFATs. All pre-exposure samples for N. caninum tested seronegative for both organisms. All postexposure samples that were seropositive for N. caninum were also positive for N. hughesi, although N. hughesi antibody titers were usually 1 dilution lower (P < 0.02). Serologic surveys for N. caninum may be confounded by cross-reacting titers with N. hughesi, but true positive N. caninum antibody titers are greater than, or equal to, cross-reacting N. hughesi antibody titers.

Serum samples from 419 polar bears (Ursus maritimus) from Svalbard and the Barents Sea (collected 1990–2000) and 108 polar bears from East Greenland (collected 1999–2004) were assayed for antibodies against Toxoplasma gondii using the modified agglutination test. Antibody prevalences were 3.6% among cubs dependent on their mothers and 21.4% among subadults and adults. Among subadults and adults there was an interaction between population and sex, with similar prevalences among females (Svalbard = 19.5%, Greenland = 18.0%), but a high frequency among Svalbard males (28.7%) as compared to Greenland males (5.8%). The pattern was also significant after correcting for differences in age distribution. The sex-population interaction term is believed to be connected to area- and sex-specific feeding ecology. The prevalences of antibodies against T. gondii in Svalbard and Greenland were high compared to previously reported findings in polar bears from Russian and Alaskan areas.

Pigs are considered to be the most important meat source of Toxoplasma gondii for humans in the United States. In the present study, 168 T. gondii isolates (designated TgPgUs15-182) from various sources were genotyped using 10 polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). Genotyping data from an additional 14 isolates collected from T. gondii–infected pigs in Maryland were included for analysis. Nine genotypes (1–9) were recognized from the 182 T. gondii isolates. Most (56%, 102) isolates were clonal Type II (genotypes 1 and 2) and 27% (49) were clonal Type III (genotype 3) strains. Genotype 4 had Type II alleles, with the exception of Type I alleles at loci Apico and L358. Eight isolates (genotype 5) from Iowa had a combination of alleles I, II, and III at different loci. The remaining 6 isolates were divided into genotypes 6–9 and had a combination of different alleles. Eight of the 9 genotypes were previously reported in different animal species and geographic regions. In conclusion, along with the predominance of clonal Type II and III strains, a few diverse, previously unrecognized T. gondii lineages were found circulating in domestic pigs used for human consumption.

The evolution of parasitism is often accompanied by profound changes to the developmental program. However, relatively few studies have directly examined the developmental evolution of parasitic species from free-living ancestors. The lined sea anemone Edwardsiella lineata is a relatively recently evolved parasite for which closely related free-living outgroups are known, including the starlet sea anemone Nematostella vectensis. The larva of E. lineata parasitizes the ctenophore Mnemiopsis leidyi, and, once embedded in its host, the anemone assumes a novel vermiform body plan. That we might begin to understand how the developmental program of this species has been transformed during the evolution of parasitism, we characterized the gross anatomy, histology, and cnidom of the parasitic stage, post-parasitic larval stage, and adult stage of the E. lineata life cycle. The distinct parasitic stage of the life cycle differs from the post-parasitic larva with respect to overall shape, external ciliation, cnida frequency, and tissue architecture. The parasitic stage and planula both contain holotrichs, a type of cnida not previously reported in Edwardsiidae. The internal morphology of the post-parasitic planula is extremely similar to the adult morphology, with a complete set of mesenterial tissue and musculature despite this stage having little external differentiation. Finally, we observed 2 previously undocumented aspects of asexual reproduction in E. lineata: (1) the parasitic stage undergoes transverse fission via physal pinching, the first report of asexual reproduction in a pre-adult stage in the Edwardsiidae; and (2) the juvenile polyp undergoes transverse fission via polarity reversal, the first time this form of fission has been reported in E. lineata.

A putative “entomoparasitic adult form” of Bursaphelenchus luxuriosae was found in the tracheal system and body cavity of its cerambycid vector beetle, Acalolepta luxuriosa. Morphologically, this form is intermediate between the usual mycophagous adult and the phoretic dauer juvenile, i.e., it shares the primary and secondary reproductive features with the mycophagous phase but shares a degenerate digestive tract with JIV dauer juveniles. In addition, the “parasitic form” has specialized characters relative to the mycophagous phase analogous to dicyclic Hexatylina, e.g., vacuole-like dots (assumed to be sensory organs) at the anterior end, a very long vulval flap, conical female tail, and elongated male spicules. The presence of insect-parasitic juveniles has been reported in several Bursaphelenchus species, but this is the first confirmed report of a putative “entomoparasitic adult form” in the genus. Thus, the “parasitic adult form” of B. luxuriosae is hypothesized to be an autapomorphic character of this species in the genus. The physiological impact of the parasitism on the host beetle is assumed to be weak because no clear symptoms were observed in the infested beetles. Also, no nematode eggs or propagative juveniles were observed in the beetle host, suggesting that more research is needed to confirm the nature of the association (parasitic vs. endophoretic).

One of the preferred freshwater crayfish species in astaciculture is the large parastacid Cherax quadricarinatus, a native to Australia. This crayfish yields good production in warm water, and, therefore, it has been translocated from Australia to other countries. One such country is Uruguay, where the initial attempt in astaciculture is being conducted. In a Uruguayan crayfish farm, a large number of Diceratocephala boschmai (Platyhelminthes, Temnocephalida) were found on the body surface and branchial chamber of C. quadricarinatus. This is the first report of an introduced temnocephalidan in the New World, and of D. boschmai in a locality outside Australia/New Guinea. The original description of D. boschmai was based on a few characters that separated this species from the other Temnocephalida; therefore, a detailed description of the Uruguayan population of D. boschmai is also provided. The conditions in which D. boschmai is found in Uruguay are suitable for its development and reproduction. This raises concern about the effect that its release into the environment might have.

Leeches exhibit a marked scope of diversity, including different kinds of symbiosis. The aim of the present study was to demonstrate through biochemical and histological analysis that a species of piscicolid leech, Myzobdella platensis, is a true parasite of blue crabs, feeding on their hemolymph and using them as a site for cocoon deposition. In a total of 48 blue crabs collected on October 2007 at 3 sites of the São Vicente Estuary, 12 specimens were infested with leeches. Callinectes bocourti (n = 7) was the most infested species with leeches and cocoons; it was chosen for biochemical and histological assays. The immunoblotting assays showed a positive reaction of the proteins in the intestinal samples of leeches collected from crabs using antihemocyanin polyclonal antibody of Ampullaria canaliculata. In addition, leech intestinal samples were recognized by antihemolymph polyclonal antibody of nonparasitized blue crabs. Histological sections of leech gut showed hemocytes and a granular matrix similar to those found in crab blood vessels. Collectively, this evidence strongly suggests a parasitic interaction between the leech M. platensis and the blue crab C. bocourti, in which the former utilizes the latter as a site for cocoon deposition and possibly for dispersal similar to that proposed for Myzobdella lugubris in Callinectes sapidus in North America.

The bufflehead duck (Bucephala albeola) (Mergini) ranges from Alaska and Canada to the United States and Mexico. Buffleheads ingest invertebrates as a major component of their food throughout life. Puddle ducks (Anatinae) change mainly to vegetable foods at an early age. Loss of helminth parasite species and abundance in the fall migration has been reported for puddle ducks. This change in food has been hypothesized to cause some of the parasite loss. Nothing was known about helminth community dynamics in wintering buffleheads, including possible loss of helminths evident in puddle ducks. Helminth communities of the buffleheads were compared between wintering localities in the Chihuahua Desert of Texas and Mexico and to results from a nesting locality in Manitoba, Canada. There were no significant differences for average helminth species richness, prevalence, or abundance between the nesting and wintering grounds. Replenishment of helminths occurred in wintering buffleheads, and abundance of some helminth species increased significantly over time. The majority of helminth life cycles were indirect. These results were mostly related to the bufflehead's continued ingestion of invertebrate foods. Seven of the 41 helminth species present in buffleheads were in common to the 3 localities. Helminth community similarities between Manitoba and Texas and Manitoba and Mexico were much lower than similarity between Texas and Mexico. The dissimilarities were mostly attributable to differences in habitats but may also indicate loss of helminth species with subsequent infections with other species. The component community of the bufflehead from the Chihuahua Desert in Texas (n = 41) consisted of 23 species of helminths (10 species of cestodes, 7 trematodes, 4 nematodes, and 2 acanthocephalans) for a total of 4,008 individual specimens. Cestodes accounted for 69% of the total abundance. The component community of B. albeola from the Chihuahua Desert in Mexico (n = 26) consisted of 18 species of helminths (7 species of cestodes, 5 trematodes, 4 nematodes, and 2 acanthocephalans) for a total of 4,345 individual specimens. Cestodes accounted for 83% of the total abundance. The bufflehead shared 10 helminth species with other species of waterfowl utilizing habitats in the Chihuahua Desert. A checklist of all helminth parasites reported for buffleheads includes 27 identified to genera, 40 to species, and 12 species from buffleheads from the Chihuahua Desert in common with buffleheads from other localities.

We have previously shown that parasite eggs have been identified in the coprolites of Korean mummies. These eggs have shed light on parasitic infection patterns in Korean populations living several hundred years ago. We conducted a scanning electron microscopy (SEM) study on ancient Trichuris trichiura, Ascaris lumbricoides, Metagonimus yokogawai, Paragonimus westermani, and Gymnophalloides seoi eggs recovered from Korean mummies of the Joseon Dynasty. We anticipated that the taphonomic conditions of mummification would alter the eggs of certain species but not of others. Our SEM data show that each species of ancient egg exhibited different degrees of preservation. Thus, some of them, for example, M. yokogawai, exhibited a better preservation status than others, suggesting that they should be the first candidates considered when choosing subjects for future paleoparasitological studies.

Falcaustra batrachiensis n. sp. (Ascaridida, Kathlaniidae) from the large intestine of Nyctimystes cheesmani (Anura, Hylidae) is described and illustrated. Falcaustra batrachiensis represents the fifth Australo-Papuan species assigned to the genus and is distinguished from other Australo-Papuan species by the distribution pattern of the caudal papillae (8 precloacal, 2 adcloacal, 6 postcloacal, and 1 median), length of spicules (1.4–1.6 mm) and presence of pseudosuckers (5 or 6). Nyctimystes cheesmani was found to harbor 7 additional species of Nematoda, adults of Cosmocerca novaeguineae, Maxvachonia ewersi, Meteterakis crombiei, Physalopteroides milnensis, Pseudorictularia dipsarilis, Seuratascaris numidica, larvae in cysts of Abbreviata sp., and 1 species of Acanthocephala, Acanthocephalus bufonis. Nyctimystes cheesmani represents a new host record for each of these helminth species.

During a parasitological survey of Galaxias maculatus (Jenyns) from Patagonian Andean Lakes, specimens of a new species of nematode were collected from the stomach of fish and studied by light and scanning electron microscopy. This species is described as the only member of a newly proposed genus of Cystidicolidae by having oral opening dorsoventrally elongated, 2 lateral pseudolabia, 4 cephalic papillae, stoma (vestibule) elongated, esophagus divided into anterior muscular and posterior glandular sections, and caudal alae in males. Placonema n. gen. (Habronematoidea, Cystidicolidae) is characterized by the combination of the following features: oral opening dorsoventrally elongated demarcated by 4 sclerotized plates and 2 well-developed pseudolabia projected to the buccal cavity, each pseudolabium with conspicuous, conical, anterior protuberances. Four cephalic papillae and deirids simple. Male with caudal alae, area rugosa absent, 4 pairs of preanal papillae, unpaired papilla present on anterior cloacal lip, and 6 pairs of postanal papillae. Larvigerous eggs without filaments. Placonema pataguense n. gen. n. sp. infects the stomach of G. maculatus from Lake Patagua (Patagonia, Argentina) and is the first species of Cystidicolidae described from G. maculatus.

Blocks of frozen lungs of 2 Australian fur seals (Arctocephalus pusillus doriferus Wood Jones, 1925), 2 New Zealand fur seals (A. forsteri [Lesson, 1828]), and 1 sub-Antarctic fur seal (A. tropicalis [Gray, 1872]) from 3 different locations (Australia, New Zealand, and South Africa, respectively) were examined and found to contain lung parasites. This represents the first thorough description and identification of a new species, Parafilaroides normani, from an eared seal (Otariidae) in the Southern Hemisphere. The new species is described, illustrated, and differentiated from the 6 recognized species in the genus by body size, spicule shape and size, vulva to anus length, and vaginal sphincter musculature.

A new proteocephalidean cestode is described from 2 catfishes, Clarias gariepinus (type host) and C. cf. anguillaris (Siluriformes: Clariidae), from Ethiopia (type locality), Sudan, Tanzania, and Zimbabwe, and a new genus, Barsonella, is proposed to accommodate it. The genus belongs to the Proteocephalinae because its genital organs (testes, ovary, vitellarium, and uterus) are situated in the medulla. Barsonella lafoni, the type and only species of the new genus, is characterized mainly by the possession of an additional opening of each sucker; circular musculature on the anterior margin of suckers, serving as a sphincter; a small thin-walled glandular apical organ; absence of well-developed osmoregulatory canals in mature, pregravid, and gravid proglottids; and a large strobila, up to 173 mm long and 3.2 mm wide. Species of Marsypocephalus Wedl, 1861 (Marsypocephalinae), other large-sized proteocephalidean tapeworms occurring sympatrically in African catfishes (Clarias and Heterobranchus) and also possessing a sphincter-like, circular musculature on the anterior part of suckers, differ from B. lafoni in the absence of an additional sucker opening and glandular apical organ, the cortical position of the testes, well-developed osmoregulatory canals throughout the strobila, and a large cirrus sac. Proteocephalus glanduligerus (Janicki, 1928), another cestode parasitic in Clarias spp. in Africa, is much smaller than B. lafoni (maximum length 15 mm), has suckers without additional opening and circular musculature on the suckers, a large-sized glandular organ, much larger than suckers, and well-developed osmoregulatory canals. Comparison of partial sequences of the 28S rRNA gene for 7 samples of B. lafoni from 2 different hosts and 4 localities in Ethiopia, Sudan, and Tanzania has shown a very low genetic variability. In a limited phylogenetic analysis, B. lafoni formed a clade with Corallobothrium solidum Fritsch, 1886 (Proteocephalidae: Corallobothriinae), an African electric catfish parasite. This clade was the sister group of almost all Neotropical taxa from pimelodid and other catfishes.

Numerous microscopic studies of coccidian oocysts from avian feces have become the basis for species identification. In contrast, molecular studies of wild birds' Coccidia are still in their infancy and are mostly based on DNA extracted from the blood stages of these parasites. Linking microscopic and molecular data requires a method that reliably extracts DNA from single oocysts with parallel detailed morphological examination of the same cell. We offer a thorough manual of isolating, photographing, and trapping single oocysts from avian feces, followed by extraction of parasite DNA and amplification of mitochondrial DNA from the same cells. In 39 single oocysts from 6 wild blackcaps, we combined microscopic studies of individual cells with studies on their mitochondrial haplotype. In 72% of the single oocysts sampled, we detected unambiguous sequences. From feces and blood of investigated birds, we obtained 6 different haplotypes of Isospora sp. (iSAT1-iSAT 6), finding both the same haplotype in different host individuals and various haplotypes in the same host individual. Our described methodology enables linking the huge amount of morphological data with innovative gene analysis. This method expands the scope of genetic studies conducted on Isospora species, including routine molecular analysis of single oocysts isolated from fecal samples.

Six specimens representing a new genus of cyclocoelid, Psophiatrema greineri n. gen., n. sp. (Digenea: Cyclocoelidae: Cyclocoelinae), were collected from the air sacs of a common or grey-winged trumpeter, Psophia crepitans (Gruiformes: Psophiidae), from Guyana on 19 January 2001. The bird was being maintained in an aviary at Emerald Forest Bird Gardens, Fallbrook, California. The new genus has an intertesticular ovary forming a triangle with the testes, placing it in Cyclocoelinae. There are currently 2 genera recognized in Cyclocoelinae: Cyclocoelum and Selfcoelum. The new genus is most similar to Selfcoelum by having a postpharyngeal genital pore, but differs from both Cyclocoelum and Selfcoelum by having the vitelline fields confluent posteriorly. A revised key to the Cyclocoelinae is provided to include the new genus, and the subfamily diagnosis is emended to reflect that the vitelline fields can be either confluent or not posteriorly.

Species of Pseudocorynosoma are North and South American acanthocephalans that use waterfowl as definitive hosts and amphipods as intermediate hosts, whereas species of Andracantha occur in fish-eating birds with a worldwide distribution. Pseudocorynosoma and Andracantha were originally described as Corynosoma (now restricted to endoparasites of marine mammals). Morphologically, Andracantha is distinct from other genera of Polymorphidae in possessing 2 fields of spines on the trunk, whereas Corynosoma and Pseudocorynosoma have a single field. A recent phylogenetic hypothesis based on morphological characters suggested that Andracantha is closely related to Corynosoma, whereas Pseudocorynosoma was of uncertain phylogenetic position within the Polymorphidae. To test the systematic affinities of these 3 genera, we sequenced 2 nuclear genes (SSU and LSU ribosomal DNA) and 1 mitochondrial gene (cytochrome c oxidase subunit 1; cox 1) of species representing Corynosoma, Andracantha, and Pseudocorynosoma and analyzed the data, including available sequences of other polymorphids. Maximum parsimony (MP), maximum likelihood (ML), and Bayesian analyses of the combined (SSU LSU) sequences and the concatenated data of 3 genes (SSU LSU cox 1) placed Andracantha as the sister taxon to Corynosoma with robust support values. All analyses also showed that Pseudocorynosoma is an independent lineage that does not share a common ancestry with Andracantha and Corynosoma. These phylogenetic hypotheses suggest that birds were the ancestral hosts of polymorphids and that the association of Corynosoma with marine mammals represents a subsequent episode of colonization.

Falcaustra odaiensis n. sp. (Nematoda: Cosmocercoidea: Kathlaniidae) is described from the Japanese clawed salamander, Onychodactylus japonicus (Houttuyn, 1782), and Ishizuchi salamander, Hynobius hirosei Lantz, 1931 (Caudata: Hynobiidae), in Japan. This nematode is characterized by having multiple pseudosuckers in the male. It resembles Falcaustra araxiana, Falcaustra duyagi, Falcaustra lambdiensis, Falcaustra pillaii, Falcaustra siamensis, and Falcaustra washingtonensis, but is readily distinguished by the spicule length and/or egg size. This is the first Falcaustra species described from Japanese salamanders.

During the winter of 2004, 48 fecal samples were collected from live-trapped fox squirrels (Sciurus niger) from central Wyoming (Natrona County) and examined for species of Eimeria. Two species, Eimeria lancasterensis (prevalence, 65%) and Eimeria ontarioensis (prevalence, 27%), were identified. Genomic DNA sequences ITS1 and ITS2 were amplified, cloned, and sequenced. Additional sequences from E. lancasterensis isolated from a Delmarva fox squirrel (Sciurus niger cinereus) collected on Chincoteague Island, Virginia, were also identified. Comparison of pairwise distances suggests that E. lancasterensis from Wyoming and Virginia are conspecific. Maximum Parsimony tree construction identified 2 lineages, one E. ontarioensis and one E. lancasterensis; and both lineages had a strong bootstrap support (100%). The Maximum Parsimony analysis was unable to resolve the Wyoming and Virginia strains.

Some microsporidian parasites belonging to the genus Nosema infect bees. Previous phylogenies of these parasites have produced alternative, conflicting relationships. We analyzed separately, and in combination, large and small subunit ribosomal DNA sequences of Nosema species infecting bees under neighbor-joining, maximum parsimony, maximum likelihood, and Bayesian frameworks. We observed a sister relationship between Nosema ceranae and Nosema bombi, with Nosema apis as a basal member to this group. When compared to their respective hosts (Apis cerana, Bombus spp., and A. mellifera), 2 plausible evolutionary scenarios emerged. The first hypothesis involves a common ancestor of N. bombi host-switching from a historical Bombus lineage to A. cerana. The second suggests an ancestral N. ceranae host-switching to a species of Bombus. The reported events offer insight into the evolutionary history of these organisms and may explain host specificity and virulence of Nosema in these economically important insects.

Haplorchis popelkae n. sp. is described from the intestine of the short-necked turtles Elseya dentata and Emydura victoriae in Northern Territory, Australia. This slipper-shaped heterophyid digenean differs from the 9 previously known species of Haplorchis in its larger overall size, a forebody that is wider than the hindbody, comparatively short ceca, and a ventral sucker that is less heavily spined than in many other species in the genus. This is the first report of Haplorchis in turtles.

Haemogregarina macrochelysi n. sp. (Apicomplexa: Haemogregarinidae) of the alligator snapping turtle, Macrochelys temminckii, is characterized by slender, recurved gamonts 29–35 × 3–4.5 μm, in which the anterior limb comprises 48–54% of the total length. The gamont nucleus, 5–7.5 × 2–5 μm, is situated at approximately midbody of the gamont. Meronts typical of Haemogregarina occupying erythrocytes have 3-8 small, compact nuclei and are 13–17 × 4.5–9 μm. Erythrocytic meronts that contain larger, nearly square or rectangular nuclei become rounded, and then undergo 7 or more nuclear divisions, which produce very large, usually ovoid to rounded meronts that may contain up to 150 nuclei or more within the thinly stretched host erythrocyte membrane. In tissues of the Placobdella spp. leech vectors, merogony occurs directly from sporozoites, forming merozoites that presumably are infective for the turtle host.

We searched the National Cancer Institute (NCI) compound library for structures related to the antitumor quinoline NSC3852 (5-nitroso-8-quinolinol) and used a computer algorithm to predict the antiprotozoan activity for each of 13 structures. Half of these compounds inhibited Toxoplasma gondii tachyzoite propagation in human fibroblasts at ≤1 μM. The active compounds comprise a series of low-molecular-weight quinolines bearing nitrogen substituents in the ring-5 position. NSC3852 (EC₅₀ 80 nM) and NSC74949 (EC₅₀ 646 nM) were the most potent. NSC3852 also inhibited Plasmodium falciparum growth in human red blood cells (EC₅₀ 1.3 μM). To investigate the mechanism for NSC3852's anti–T. gondii activity, we used chemiluminescence assays to detect reactive oxygen species (ROS) formation in freshly isolated tachyzoites and in infected host cells; the absence of ROS generation by NSC3852 in these assays indicated NSC3852 does not redox cycle in T. gondii. Inhibitors of enzyme sources of free radicals such as superoxide anion, nitric oxide (NO), and their reaction product peroxynitrite did not interfere with the anti–T. gondii activity of NSC3852. However, inhibition of T. gondii tachyzoite propagation by NSC3852 involved redox reactions because tachyzoites were protected from NSC3852 by inclusion of the cell permeant superoxide dismutase mimetic, MnTMPyP, or N-acetylcysteine in the culture medium. We conclude that the Prediction of Activity Spectra for Substances (PASS) computer program is useful in finding new compounds that inhibit T. gondii tachyzoites in vitro and that NSC3852 is a potent T. gondii inhibitor that acts by indirect generation of oxidative stress in T. gondii.

Outside its native range, the invasive New Zealand mud snail (NZMS), Potamopyrgus antipodarum, is rarely reported to harbor parasites. To test this observation, 7 sites along the Snake River and Polecat Creek in the Grand Teton National Park/John D Rockefeller Memorial Parkway area (Wyoming) were surveyed for native aquatic snails, NZMS, and associated digenean trematodes, in July 2005. At 6 sites, native snails harbored patent digenean infections; within 2 hr, ≤10% of lymnaeid snails shed furcocercariae or xiphidiocercariae, and ≤42% of physid snails released furcocercariae or echinostome cercariae. Partial 18S rDNA sequences were recovered from several furcocercariae. Potamopyrgus antipodarum was present at, and collected from, 5 sites. Polymerase chain reaction assays targeting digenean rDNA sequences in DNA extracted from pools of 150 NZMS snails did not detect parasites. The examination of 960 NZMS by overnight shedding yielded 1 occurrence of (surface-encysted) metacercariae of an unclassified notocotylid (based on 18S and 28S rDNA sequences). The dissection of 150 ethanol-fixed NZMS (30/site) revealed 2 types of digenean metacercariae encysted in tissues of 5 snails from Polecat Creek. Thus, invasive NZMS may serve as first and second intermediate host for digenean parasites.

The spleen is an important organ of vertebrates. Splenic mass can change in response to a variety of factors. We tested whether splenic mass of masked shrews, Sorex cinereus, was related to sex, age, time of the year, or intensity of bladder nematode (Liniscus [=Capillaria] maseri) infection, after controlling for host body mass. For females, body mass was a strong predictor of splenic mass. For males, splenic masses were greater later in the year and in more heavily infected males. The latter appeared to represent a threshold response wherein only the most heavily infected individuals had enlarged spleens.

Eimeria species are frequently found in water buffalo (Bubalus bubalis) in Brazil. Here, we report those Eimeria spp. that infect buffalos during their first year of life. Fresh fecal samples were examined from 2 groups (1 group/yr for 2 yr, 2000–2002), each with 18 water buffalo calves (both sexes), from birth through 12 mo of age, in Selvíria, MS, Brazil. Five oocyst morphotypes were observed, i.e., Eimeria ellipsoidalis and Eimeria zuernii, both previously described from water buffalo, and 3 other morphotypes consistent with descriptions of known Eimeria spp. from Artiodactyla hosts, but originally described from other genera than those in which we found them (referred to here as Eimeria species 1–3). Our results showed that buffalo calves started shedding oocysts in their feces between 6–29 days of age, with the highest concentration ranging from 188–292 oocysts/g of feces. The 3 unnamed oocyst morphotypes in the calf feces resembled E. auburnensis (Eimeria sp. 3), E. cylindrica (Eimeria sp. 1), and E. subspherica (Eimeria sp. 2). The most prevalent species were Eimeria sp. 1 and E. ellipsoidalis, which dominated in the youngest animals (6 to 133 days old). Eimeria zuernii oocysts, in contrast, were found only in low numbers in the feces of older calves (208 to 283 days old). Calves were infected more frequently during the rainy season (September to January) in both years, but cows were negative for Eimeria spp., whenever feces were collected (spring, winter, autumn, or summer seasons).

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 152 free-range chickens (Gallus domesticus) from 22 municipalities in 7 northeastern states (Pernambuco, Rio Grande do Norte, Maranhão, Bahia, Ceará, Sergipe, and Alagoas) of Brazil was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT); 81 (53.3 %) chickens had titers of 1:5 in 26, 1:10 in 9, 1:20 in 4, 1:40 in 1, 1:80 in 6, 1:160 in 6, 1:320 in 13, 1:640 in 6, 1:1,280 in 3, 1:2,560 in 6, and 1:5,120 or higher in 1. Hearts and brains of 81 seropositive chickens were bioassayed individually in mice. Toxoplasma gondii was isolated from 23 chickens with MAT titers of 1:5 or higher; the isolates were designated TgCKBr165-187. Five isolates killed all infected mice. Results indicate widespread contamination of rural environment in Brazil with T. gondii oocysts.

Two freshly-dead female Florida panther (FP) neonates, Puma concolor couguar (=Puma concolor coryi), an 11-day-old and a 17-day-old, were collected in the Florida Panther National Wildlife Refuge (26°14′N, 81°36′W), Collier County, Florida. The 2 neonates were siblings and had presumably fed only on milk from the dam since birth. A 12-day-old female FP neonate was collected in the Big Cypress National Preserve (26°05′N, 81°15′W), Collier County, Florida and had also fed only on milk from the dam since birth. Milk was the only food item found in the gastrointestinal tract of these neonates. Mesocercariae and diplostomula of Alaria marcianae were collected from the lungs of the 3 neonates, indicating a transmammary route of infection. No mesocercariae, diplostomula, or mature A. marcianae were seen in the stomach or small intestine. The probable paratenic host for the A. marcianae infection in the adult Florida panther is the raccoon (Procyon lotor).

In previous studies, we have reported that the larval products (crude extract [CE] and excretory-secretory [ES]) of Anisakis simplex showed a dose-dependent inhibition of the lysis mediated by classical (CP) and alternative pathways (AP) of the human complement system, with the major inhibition on the CP rather than on AP. This inhibition of hemolysis is due to the consumption of complement factors because the assays performed shortening the preincubation period result in a significant decrease of the inhibitory effect on the lysis of the larval products compared with the standard time. Likewise, we found that the larval products reduce the inhibitory percentages in the CP using C3-deficient sera, but not in the AP, which could indicate that other complement components are implicated in the inhibitory effect in the CP. Hence, we have studied the activity of the larval products of A. simplex on individual components in the CP, using different complement-deficient sera. The investigated complement molecules were C1q, C2, C4, C5, C6, C7, C8, and C9. The larval products showed activity at the C2 level but failed to have a significant effect on the other components. Therefore, CE and ES products from A. simplex interact with C3 and C2 complement proteins, which are early components of the complement system, but not with the late complement components.

Gordiid larvae enter and encyst within a large variety of aquatic vertebrates and invertebrates. Cysts of a single species of gordiid have been found in such diverse hosts as insect larvae, snails, crustaceans, fish, and many others. One particularly surprising host report was made nearly 100 yr ago, i.e., cysts within adult trematodes. The origin of these cysts remained unclear, but it was suggested that the gordiid larvae gained entry into the adult flukes via their definitive, vertebrate host. In the present study, the finding is reported of gordiid cysts within monostome redia within a Physa gyrina snail. This is the first time gordiid cysts have been reported within redia, and this provides an alternative explanation for the presence of cysts within adult trematodes.

We compared parasite load (prevalence and mean intensity) of Eutrombicula alfreddugesi larvae on the lizard Liolaemus tenuis sampled during January 2006 and 2007 from the interior and edges of large forest tracts in the coastal Maulino Forest (35°59′S, 72°41′W) and from nearby forest fragments (1.5–20 ha). All lizards were parasitized by chiggers regardless of location (prevalence, 100%); however, mean intensity of infestation was significantly lower at forest fragment edges compared with either large forest interiors or forest edges. We attribute differences in mean intensity to differences in microclimate among localities; maximum air temperature was significantly higher and relative humidity significantly lower in fragment edges compared with either large forest tract interior or edges.

Human infections with Gnathostoma spinigerum frequently occur in southern Vietnam. Previous investigations have implicated infected swamp eels (Monopterus albus) as an important source of infection to humans. Because aquaculture of M. alba is an important farming activity in Vietnam, a 2-yr study was carried out to assess the relative importance of farmed and wild eels as potential sources of gnathostome infections in humans. Eels sold for public consumption in markets in southern provinces were examined between November 2005 and August 2007. Although no infections were detected in 1,020 cultured eels and 1,021 wild-caught eels collected from November 2005 to September 2006, larval G. spinigerum (AL3) infections were first detected in September 2006 in 28 of 230 wild-caught eels (12.2%) obtained from markets in Long An province and the Hoc Mon district of Ho Chi Minh City. Subsequently, monthly surveillance of wild-caught eels from these markets was carried out through August 2007. Prevalence of AL3 varied monthly, ranging from 0.8 to 19.6%. Both prevalence and infection intensity were higher during the latter part of the rainy season (August– October). These results demonstrate that potentially zoonotic G. spinigerum larvae are common in wild eels in southern Vietnam and present a risk to consumers of raw fish dishes, especially during the annual rainy season. This information could help target public health education efforts in the region. The basis for the seasonal variation on eel infections is not known, but may be related to climate effects (flooding, higher temperatures) on intermediate host species ecology.

The helminth fauna of pelagic-stage loggerhead sea turtles, Caretta caretta, is still poorly known. Here, we describe the helminth-component community of healthy, free-ranging juvenile loggerhead sea turtles captured in the waters around Madeira Island, Portugal. Fifty-seven were used in this study. The esophagus, stomach, intestine, liver, gallbladder, spleen, kidneys, trachea, bronchi, urinary bladder, heart, left and right aortas, and coelomic cavity were macroscopically inspected; organs and tissues were removed and washed through a sieve. A search for parasites was made using a stereoscopic microscope; recovered parasites were fixed and stored in 70% alcohol until staining and identification. Prevalence, mean intensity, and mean abundance values were recorded. In total, 156 parasite specimens belonging to 9 species were found: nematodes included Anisakis simplex s.l. (larvae) and an unidentified species; digenetic trematodes present were Enodiotrema megachondrus, Rhytidodes gelatinosus, Pyelosomum renicapite, and Calycodes anthos; acanthocephalans included Bolbosoma vasculosum and Rhadinorhynchus pristis; a single cestode, Nybelinia sp., was present. Parasite infections were found to have both low prevalences and intensities. Possible reasons for this include the oligotrophic conditions of the pelagic habitat around Madeira; a ‘dilution effect’ because of the vastness of the area; and the small size, and thus ingestion rate, of the turtles. Results are discussed in terms of the various turtle populations that may use the waters surrounding Madeira. This work provides valuable information on the parasite fauna of a poorly known stage in the life of loggerhead sea turtles, thereby filling a fundamental gap with regard to features of the parasite fauna in this species.

Multiple displacement amplification (MDA) using Phi29 has proved to be an efficient, high-fidelity method for whole genome amplification in many organisms. This project was designed to evaluate this approach for use with the malaria parasite Plasmodium falciparum. In particular, we were concerned that the AT richness and presence of contaminating human DNA could limit efficiency of MDA in this system. We amplified 60 DNA samples using phi29 and scored 14 microsatellites, 9 single-nucleotide polymorphisms (SNPs), and gene copy number at GTP-cyclohydrolase I both before and after MDA. We observed 100% concordance in 829 microsatellite genotypes and in 499 SNP genotypes. Furthermore, copy number estimates for the GTP-cyclohydrolase I gene were correlated (r² = 0.67) in pre- and postamplification samples. These data confirm that MDA permits scoring of a range of different types of polymorphisms in P. falciparum malaria and can be used to extend the life of valuable DNA stocks.

During 2001 to 2004, 4 species of hawks (Buteo and Accipiter spp.) from Georgia were surveyed for Sarcocystis spp. infections by examining intestinal sections. In total, 159 of 238 (66.8%) hawks examined were infected with Sarcocystis spp. Samples from 10 birds were characterized by sequence analysis of a portion of the 18S rRNA gene (783 base pairs). Only 3 of the 10 sequences from the hawks were identical; the remainder differed by at least 1 nucleotide. Phylogenetic analysis failed to resolve the position of the hawk Sarcocystis species, but they were closely related several Sarcocystis species from raptors, rodents, and Sarcocystis neurona. The high genetic diversity of Sarcocystis suggests that more than 1 species infects these 4 hawk species; however, additional molecular or experimental work will be required to determine the speciation and diversity of parasites infecting these avian hosts. In addition to assisting with determining species richness of Sarcocystis in raptors, molecular analysis should be useful in the identification of potential intermediate hosts.