Histomonads grew rapidly in Dwyer's medium, consisting of medium 199, chick embryo extract, serum, and rice powder, reaching a population size of about 5 × 10⁵ in 3–4 days, followed by a rapid decline. Substitution of other cell culture media (L-15, MEM, or RPMI) for M199 was also satisfactory, except for Waymouth's medium, which produced a lower and later peak of growth. Omission of serum or rice rendered media unsuitable for growth. Bacteriological culture media did not support growth of histomonads. Media that included glucose were unsuitable because the pH of the cultures dropped to about 4. The effect of glucose on pH was due to the action of bacteria. Oxygen inhibited growth of histomonads. There was no growth when culture tubes were not capped tightly, regardless of the medium used. Histomonads grew well with rice flour, cornstarch, oat flour, rye flour, and buckwheat flour. Barley and blue corn meal were less satisfactory. It appeared that the requirements for growth of the lumen phase Histomonas meleagridis included a suitable physiological saline, serum (of any source), and a starch source (grain flour). Anaerobic conditions and a pH near neutral were best. Histomonads separated into pure cultures by flow cytometry would not grow without the inclusion of an unspecified species of bacteria.

Leishmania chagasi, a causal agent of visceral leishmaniasis, requires passage through lab animals such as hamsters to maintain its virulence. Hamster infection is typically accomplished via cardiac puncture or intraperitoneal injection, procedures accompanied by risks of increased animal stress and death. The use of the hamster model also necessitates a regular supply of infected animals, because L. chagasi parasites newly isolated from an infected hamster can be grown in culture for only several weeks before loss of function/phenotype occurs. In an effort to decrease animal usage and animal stress, experiments were performed to assess a more gentle inoculation procedure (saphenous vein inoculation) and the use of cryopreserved parasite cells for research experiments. Of 81 hamsters inoculated by the saphenous vein, 80 became infected as determined ante mortem, by display of clinical symptoms of leishmaniasis (onset of symptoms at 105 ± 22 days post-inoculation), and postmortem by the presence of parasites within the spleen. Splenic parasite load calculated for a subset (n  =  34) of infected hamsters was 124 to 26,177 Leishmania donovani infection units. Cryopreserved, and never-stored, cells were equivalent in all properties evaluated, including developmental changes in morphology during culture, culture growth rates, parasite resistance to serum-mediated lysis, and expression of developmentally regulated surface proteins major surface protease and promastigote surface antigen.

During a parasite survey of galaxiid fishes (Galaxiidae) from Patagonian Andean lakes, a new species of nematode, Hedruris suttonae n. sp. was collected from the stomach of the native Galaxias maculatus (Jenyns) and G. platei (Steindachner). Specimens were examined by light and scanning electron microscopy, especially head morphology, female caudal prehensile structure, and distribution of spines. The new species is distinguished by body and tail size, morphology and size of spicules, the arrangement of caudal papillae in the male, the female caudal hook, and size of eggs. Hyalella patagonica (Ortmann), a Neotropical species of Amphipoda, is reported as its natural intermediate host. Data regarding prevalence and mean intensity in the intermediate and definitive hosts are included. The diet and habitat of the hosts, the percentage of gravid females, the high values of prevalence, and mean intensity in galaxiid fishes, as well as the wide distribution of H. suttonae, collectively indicate that, in these oligotrophic Andean lakes, G. maculatus and G. platei are true definitive hosts of this nematode.

A Myxobolus sp., morphologically resembling M. toyamai, M. longisporus, and M. koi, was isolated from the gills of a koi, Cyprinus carpio, that died in an ornamental pond. Large plasmodia were localized within lamellae, causing severe disruption of the normal branchial architecture, sufficient to compromise respiration. Although the case isolate shared several features with the aforementioned species, several key characteristics were most compatible with M. koi. In valvular view, spores were elongate and pyriform with a rounded posterior, 15.4 (14.5–16.5) µm long and 8.3 (7.1–9.0) µm wide. Polar capsules were pyriform and elongate, 10.1 (9.0–10.9) µm long and 3.1 (2.5–3.5) µm wide. Polar filaments were coiled perpendicular to the long axis of the spore making 10 turns (9–11). A BLAST search using a generated 18S SSU rDNA sequence resulted in no direct matches. Phylogenetic analysis of the sequence indicates a close relationship to M. longisporus and M. toyamai from C. carpio, within the clade of Myxobolus species infecting gills of cyprinid fishes (89% or greater bootstrap support by maximum parsimony and minimum evolution distance analysis). Although published descriptions are inconsistent, morphometric similarities to this isolate suggest a morphotype of M. koi. The description of M. koi is supplemented here with new data on pathogenicity and spore morphology and is characterized at the molecular level using 18S small subunit rDNA sequence data.

Syphacia (Syphacia) rifaii sp. n. (Nematoda: Oxyuridae) is described from endemic Bunomys chrysocomus and Bunomys prolatus (Rodentia: Muridae) on Sulawesi Island, Indonesia. The new species is closest morphologically to Syphacia (Syphacia) sulawesiensis, parasitic in Rattus xanthurus from Sulawesi Island, by having large vesicular lateral alae in males, but is readily distinguished by having a smaller body, a round cephalic plate in both sexes, the absence of lateral alae in females, a longer relative distance between excretory pore and vulva, and smaller eggs. Syphacia (S.) rifaii is surmised to be a specific parasite of Bunomys spp. and has evolved from a common ancestor with S. (S.) sulawesiensis on Sulawesi Island.

Africanastrongylus giganticus n. sp. is described based on large ostertagiine nematodes occurring in the abomasum of African buffalo, Syncerus caffer, from Uganda; this represents the second species recognized in the genus. Specimens of A. giganticus are characterized by large size (15–19 mm in total length), a strongly tapering synlophe in the cervical region, and a great number of ridges at all levels of the body (maximum 72 attained in the third quarter); numbers of ridges exceed that reported among any known genera and species of the Ostertagiinae. We refer A. giganticus to this genus based on a strongly tapering lateral synlophe, relatively large numbers of ridges at all levels of the body, miniscule cervical papillae, poorly demarcated divisions of the ovejector, absence of vulval cuticular inflations, and the presence of slightly protruding lips at the vulva. It is distinguished from its congener, Africanastrongylus buceros, in total length, maximum number of ridges (68–72 vs. 53, respectively), structure and disposition of the synlophe, presence of strongly spiraled ovarian tracks, and eggs that are distributed in 3 or more rows in the uterus. A superficial resemblance to Longistrongylus meyeri, the only other large ostertagiine in the African fauna, is evident; these species, however, are distinct based on the synlophe and other characters. Recognition of a second species of Africanastrongylus represented by nematodes of large size suggests that prior reports of L. meyeri in Syncerus caffer may be attributable to A. giganticus.

Haemoproteus (Parahaemoproteus) cyanomitrae n. sp. (Haemosporida: Haemoproteidae) was found in the olive sunbird, Cyanomitra olivacea (Nectariniidae), in rain forests of tropical Africa. It is described based on the morphology of its blood stages and a segment of the mitochondrial cytochrome b gene (GenBank accession FJ404696), which can be used for molecular identification and diagnosis of this species. Fully grown gametocytes of new parasite are similar to many other species of haemoproteids of passeriform birds, so are not easily distinguishable from many of them at this stage of their development. Haemoproteus cyanomitrae can be readily distinguished from the majority of described species of avian haemoproteids, primarily due to its growing dumbbell-shaped gametocytes and advanced halteridial gametocytes, both of which do not touch the envelope of infected erythrocytes along their entire margin. Pigment granules are most frequently clamped in 1, or several, groups in gametocytes, with larger group of pigment granules usually located close to one end of the gametocyte; asymmetric position of pigment granules in gametocytes a characteristic feature of this parasite. Illustrations of blood stages of the new species are given, and phylogenetic analysis identifies DNA lineages closely related to this parasite, which is widespread in African rain forests, but has been recorded only in the olive sunbird so far. It is probable that H. cyanomitrae is transmitted throughout the range of the olive sunbird in Africa.

Assessing parasite specificity to vector is crucial to understanding the emergence of vector-borne diseases and the evolution of parasite diversity. Avian malaria parasites have a cosmopolitan distribution and broad avian host range, which together predict they are vector generalists, but little is known about parasite–vector associations in the wild. We tested this prediction by asking if 5 different mosquito species, known to feed on birds and abundant in the northeastern United States, were naturally infected in the field with identical avian Plasmodium spp. lineages. Mosquitoes were not pooled but rather analyzed individually, and, possibly as a result, lineage diversity was higher than reported in previous avian malaria vector studies. Plasmodium spp. lineages were rare in Aedes canadensis and absent in Aedes aurifer and Culiseta melanura. We sequenced a standard Plasmodium cytochrome b marker from Culex pipiens pipiens, Culex restuans, and Ae. canadensis. Most Plasmodium clades were shared by Cx. pipiens and Cx. restuans. In addition, 4 individual lineages were shared by both mosquito species, including the most common lineage. One Plasmodium clade, however, was only found in Cx. restuans. We therefore found limited support for our prediction that avian Plasmodium spp. vector breadth accompanies host breadth. The association of both Culex species with most Plasmodium clades, and the presence of a single parasite lineage in 3 mosquito species representing 2 genera, suggests that avian Plasmodium species are not tightly coevolved with vector species.

A new species of nematode, Pseudodelphis chilensis n. sp., is described. This parasite was found in muscles of the pectoral fins of a blenniid fish, Scartichthys viridis, from central Chile. The new species differs from the other 2 described (P. oligocotti Adamson and Roth, 1990 and P. limnicola Brugni and Viozzi, 2006) for its longer glandular esophagus and a small caecum at the beginning of the glandular esophagus; both characteristics are in males and females. In addition, the male of P. chilensis n. sp. does not have an adanal caudal papillae, and it has longer spicules than the other 2 species. This is the first Pseudodelphis species described for Chile and the second for South America.

This is the first report of Enterocytozoon bieneusi in an equid species. Feces from 195 horses from 4 locations in Colombia were examined for E. bieneusi by polymerase chain reaction. Of these, 21 horses (10.8%) were found positive for E. bieneusi. The prevalence of E. bieneusi in horses <1 yr of age was significantly higher (23.7%) than in horses >1 yr of age (2.5%). No significant differences in prevalence were observed between male (13.7%) and female horses (9%). Sequencing of the internal transcribed spacer region of the SSUrRNA locus identified 3 genotypes. Two genotypes appear to be unique to horses and were named Horse 1 and Horse 2. A third genotype, identified as genotype D, was detected in 4 horses. This genotype, previously reported to infect humans, beaver, cattle, dogs, falcons, foxes, macaques, muskrats, pigs, and raccoons, is the most ubiquitous of the E. bieneusi zoonotic genotypes. Our findings indicate that E. bieneusi from horses can be a potential source of infection for humans.

Black rats (Rattus rattus) and their stomach nematodes (Mastophorus muris) were historically introduced to islets at Palmyra Atoll in the central Pacific Line Islands. To investigate patterns of parasitism, we trapped rats and quantified nematodes on 13 islets of various sizes and habitat types. Most rats were parasitized (59%) with an average of 12 worms per infected rat. Islet size did not greatly influence parasite population biology. Nematodes also did not appear to affect rat condition (weight to skull length). The only strong and consistent factor associated with the mean abundance of nematodes in rats was habitat (dominant cover and locally dominant plant species). Thus, nematodes were much more abundant in rats from sites dominated by coconut trees (Cocos nucifera). Coconut trees may also be an introduced species at Palmyra Atoll.

The Florida kingsnake, Lampropeltis getula floridana, is host to 3 species of Hepatozoon at the type locality, Miramar, Broward County, Florida, and 2, possibly all 3, species at Cedar Key, Levy County, approximately 480 km to the northwest. Hepatozoon eurytopis, n. sp. was present also in Diadophis punctatus and Elaphe guttata in Jupiter Farms, Palm Beach County, and in Thamnophis sirtalis at Gainesville, Alachua County. Specific identity in D. punctatus was determined from gamont morphology and from sporogonic stages and gamont morphology in the infections obtained from E. guttata and T. sirtalis. Infection with all 3 species by ingestion of infected mosquitoes was easily obtained in E. guttata, but only H. eurytopis produced experimental infection in T. sirtalis and Storeria occipitomaculata. Hepatozoon eurytopis differed from the other 2 species, H. karyolysi n. sp. and H. rexi n. sp., by its short and broad, recurved gamonts with average dimensions 11.2–13.1 × 4.5–5.4 µm, length × maximum width (LW) 56–64 µm², and L/W ratios 2.13–2.80 in all host species, large round oocysts 95–303 × 91–285 µm, L/W ratios 1.03–1.05 with spherical to ovoid sporocysts 22–50 × 21–44 µm, L/W ratios 1.00–1.82 that contain 33.5–44.4 (16–76) sporozoites. The nucleus in immature gamonts is highly irregular, often fragmented in separate masses of chromatin, and commonly appears to form binucleate gamonts before maturity. Hepatozoon karyolysi has elongate, non-recurved gamonts, 15–20 × 4–7 µm, LW 64–143 µm², L/W ratios 2.3–4.5 that usually cause lysis or irregular margins of the nucleus in infected erythrocytes of the type host. Oocysts are spherical, 127–243 × 122–233 µm, L/W ratio 1.0–1.1 that contain spherical to usually ovoid sporocysts 17–30 × 17–24 µm, and produce 12–20 sporozoites. Gamonts of Hepatozoon rexi are non-recurved, 14–18 × 4–6 µm, LW 67–102 µm², and L/W ratio 2.8–4.1, and are usually surrounded by a prominent cyst wall within the host erythrocyte. Oocysts are nearly spherical, 103–178 × 103–172 µm, form usually elongate sporocysts, 30–39 × 20–25 µm L/W ratio 1.33–1.94 that contain 16–36 sporozoites. An oocyst of H. eurytopis was present in the salivary gland of an infected Aedes aegypti, which is the second report of sporogony by a Hepatozoon species occurring in this site.

The current taxonomy of leucocytozoids (Haemosporida, Leucocytozoidae) is based on the morphology of blood stages of the parasites and on limited information about their specificity. Recent molecular studies have revealed a remarkable genetic diversity of leucocytozoids, indicating that their taxonomic diversity may be greater than in the current classifications. We addressed this issue using morphological data and phylogenetic analysis of the cytochrome b gene of 14 positively identified species of avian haemosporidians. Based on the current taxonomy, Leucocytozoon toddi is the sole species of leucocytozoids parasitizing falconiform birds. However, several distinct haplotypes have been revealed in falconiform birds, suggesting that L. toddi might be a group of cryptic species. We investigated the morphology of blood stages of leucocytozoids of 2 haplotypes (2 lineages from Accipiter spp. and 3 lineages from Buteo spp.) and concluded that these parasites can be readily distinguished due to length of the cytoplasmic processes of their host cells; therefore, they do represent distinct morphospecies. Morphology of the cytoplasmic processes of host cells warrants more attention in the taxonomy of Leucocytozoon species. Leucocytozoon toddi indeed is a species group that currently includes Leucocytozoon mathisi, Leucocytozoon buteonis, and L. toddi; additional morphospecies can be added to this group in the future. It is probable that some other morphospecies of Leucocytozoon also might be groups of closely related species.

A novel coccidian parasite from the kidney of big brown bats (Eptesicus fuscus) is described. This coccidian (Nephroisospora eptesici nov. gen., n. sp.) was associated with a generally mild, focal or multifocal, well-demarcated cortical renal lesion less than 1 mm in diameter. The lesion represented cystic, dilated tubules with hypertrophied tubular epithelial cells and was present in the kidneys of 29 of 590 bats. Numerous coccidian parasites in various stages of development were present within the tubular epithelial cells and within the cyst lumina. Oocysts were collected from cystic dilated tubules. Thin-walled, sporulated ellipsoidal oocysts measuring an average of 18.9 × 20.8 µm were present in kidney tissue. The oocysts contained 2 sporocysts with 4 sporozoites. A polar body and a prominent oocyst residuum were present in the oocysts, but no micropyle, sporocyst residuum, or Stieda bodies were detected. Analysis of the 18S rRNA gene sequence put the parasite in the Sarcocystidae. The parasite is closely related to Besnoitia, Hammondia, Neospora, and Toxoplasma. Ultrastructural features, such as the presence of an apical complex in merozoites, support the identification of a coccidian. A new genus and species, Nephroisospora eptesicii, is proposed for this unusual coccidian in which the entire cycle is completed in the kidney of a single host; it has a membrane-like oocyst wall, sporogony occurs in the host rather than in the abiotic environment, and the positioning of the parasite by nucleic acid sequence indicates it to be closely allied to Sarcocystis and Besnoitia.

Terminal-spined Schistosoma sp. eggs were detected in several groups of baboons living in Kafue National Park in central Zambia. A total of 166 fecal samples was screened; egg prevalence overall ranged between 7% and 10%, while infection intensities were low. Formalin-fixed eggs had an average length of 144.5 µm and a breadth of 48.3 µm, but the schistosome species could not be unambiguously identified by size or morphology. We used molecular methods to definitively identify the parasite species. Parasite DNA was amplified from stools by polymerase chain reaction (PCR). Sequence analysis of fragments of the first internal transcribed spacer (ITS-1), mitochondrial 12S rDNA, NADH dehydrogenase subunit 6 (nad6), and cytochrome C oxidase subunit 1 (cox1) from 3 egg-positive samples revealed the presence of S. mattheei in these samples. This is the first molecular identification of S. mattheei from free-ranging baboons. Schistosoma mattheei is typically a parasite of bovids, but it can also infect humans. Schistosoma mattheei in baboons in Zambia may affect other wildlife species and humans that live in close proximity to baboons.

Two anglerfish species, Lophius piscatorius and Lophius budegassa, caught off the north and northwest coast of Spain in 2007, were examined for parasite infections. Four parasite taxa were chosen for study, i.e., Anisakis larvae Type I (sensu Berland, 1961), Hysterothylacium rigidum (Nematoda), Spraguea lophii (Microsporidia), and Chondracanthus lophii (Copepoda). Prevalence, intensity, and abundance were determined and related to host size and sex. The abundance of Anisakis larvae Type I significantly increased with size in both anglerfish species, and C. lophii was also more abundant in larger L. piscatorius specimens. The abundance of H. rigidum and the prevalence of S. lophii were unrelated to host size. The infestation with the parasites studied was not affected by host sex.

To evaluate the seasonal prevalence of human intestinal parasites in the western states of Colorado, Utah, New Mexico, and Montana, fecal samples were examined as part of routine diagnostic testing from patients experiencing gastrointestinal discomfort in August (summer) 2006, January (winter), and April (spring) 2007. Parasite identification in positive samples was confirmed using light microscopy after wet mount and trichrome staining techniques. Seventy-eight of the 1,083 patients surveyed (7.2%) in August tested positive for at least 1 species of intestinal parasite. Forty-eight of 726 (6.6%) patients and 51 of 795 (6.4%) patients tested positive for at least 1 species in January and April, respectively. Blastocystis sp. was the most prevalent, followed by Giardia lamblia. Approximately 25% of the parasite occurrences were multiple infections involving fecal-oral transmitted species. Co-infections with Entamoeba spp. and Blastocystis sp. were common, suggesting a possible fecal-oral transmission for the latter parasite. Entamoeba spp. were more likely to co-occur than independently. Other species detected included Endolimax nana, Diphyllobothrium latum, Hymenolepis nana, Dientamoeba fragilis, and Iodamoeba butschlii.

The aim of this study was to identify risk factors associated with Neospora caninum seropositivity in sheep from the State of Alagoas, in the northeast region of Brazil. Twenty-six herds were selected, and blood samples were collected from 343 animals > 6-mo-old. The indirect fluorescent antibody test (IFAT) was used for serological diagnosis of N. caninum infection. Epidemiological questionnaires were applied for each herd to identify the risk factors associated with infection. Thirty-three (9.6%) animals tested positive for anti–N. caninum IgG antibodies, with titers ranging from 1∶50 to 1∶1,600. Fourteen herds (53.8%) presented at least 1 seropositive animal. Risk factors identified were property size ≤ 30 ha (odds ratio  =  7.23; 95% CI  =  1.99–26.49) and source of water (wells municipal water courses) (odds ratio  =  4.76; 95% CI  =  1.23–18.47). This is the first ezootiological study of N. caninum infection in sheep to be carried out in the State of Alagoas. Preventive measures focusing primarily on water treatment should be taken to reduce the risk of infection by N. caninum.

The primary aim of the present study was to conduct a 1-yr parasitological survey of a Mytilus galloprovincialis mussel population from the Aveiro Estuary, Portugal. In addition, we attempted to assess the host response to parasites by examining host histopathology, including an evaluation of hemocyte infiltration and parasite encapsulation. The parasites present (prevalence) included the following: 1 protozoan, Nematopsis sp. (70%); 1 turbellarian, Urastoma cyprinae, (39%); 2 trematodes, Diphtherostomum sp. (58%) and Prosorhynchus crucibulum (0.3%); 3 crustaceans, Bathylaophonte azorica (0.3%); and 2 species of Mytilicola sp. (3.5%), M. intestinalis and M. orientalis. Highest prevalences for the most abundant species occurred in summer and autumn. Histological sections of the mantle of infected mussels revealed a marked reduction in the reproductive tissue, indicating that P. crucibulum invades the mussel and promotes a severe damage to its reproductive capacity.

Parasitic castration may result from manipulation of host energy allocation away from reproduction, which should result in castration of lightly infected hosts as well as heavily infected ones. Castration also may result from nutrient theft alone, which incidentally influences host energy allocation to reproduction and should cause reproduction to end in heavily infected hosts. Although the pseudophyllidean cestode Schistocephalus pungitii is a castrator of ninespine stickleback fish (Pungitius pungitius), the cause and significance of castration remain unknown. We used predictions about reproductive capacity in lightly and heavily infected hosts and host size (age) at which castration occurs to address these questions. In Airolo Lake, Alaska, we found that inhibition of reproduction in ninespine stickleback occurs in small fish before the size (age) of sexual maturity and is not instantaneous. Castration was observed in lightly and heavily infected adult-size hosts. Infected fish do not reach the same size as uninfected fish, possibly because of growth inhibition or selective predation. Castration in infected adult-size female ninespine stickleback appears to be the result of adaptive manipulation of host energy allocation by the parasite, but it could result from an energy budget readily influenced by parasitism.

We examined stool samples for trophozoites of the entodiniomorphid ciliate Troglodytella abrassarti Brumpt and Joyeux, 1912, from a habituated group of chimpanzees (Pan troglodytes schweinfurthii) at Mahale Mountains National Park in western Tanzania. In our study, fresh fecal samples from identified individuals were collected immediately after defecation and fixed in 10% formalin. In total, 52 samples from 38 chimpanzees (61% of 62 chimpanzees in the group) were examined using a direct smear method. A stool sample from an individual collection date from an individual chimpanzee was examined up to 3 separate times before it was called negative. Forty-eight (92%) of the 52 samples were positive, and stools from 37 (97%) of the 38 chimpanzees were positive for trophozoites of T. abrassarti. The high prevalence of T. abrassarti in these chimpanzees is consistent with previous reports of this organism in chimpanzees.

Spatial heterogeneity influences the distribution, prevalence, and diversity of haemosporidian parasites. Previous studies have found complex patterns of prevalence with respect to habitat characteristics and parasite genotype, and their interactions, but there is little information regarding how parasitemia intensity and the prevalence of co-infections may vary in space. Here, using both molecular methods and microscopy, we report an analysis of the variation of parasitemia intensity and co-infections of avian haemosporidian parasites (Plasmodium and Haemoproteus species) in 2 common African birds species, the yellow-whiskered greenbul (Andropadus latirostris) and the olive sunbird (Cyanomitra olivacea), at 3 sites with distinct habitat characteristics in Ghana. First, we found an interaction between the site and host species for the prevalence of Plasmodium spp. and Haemoproteus spp. For the olive sunbird, the prevalence of Plasmodium spp., as well as the number of individuals with co-infections, varied significantly among the sites, but these measures remained constant for the yellow-whiskered greenbul. In addition, yellow-whiskered greenbuls infected with Haemoproteus spp. were found only at 1 site. Furthermore, for both bird species, the parasitemia intensity of Plasmodium spp. varied significantly among the 3 sites, but with opposing trends. These results suggest that spatial heterogeneity differently affects haemosporidian infection parameters in these vertebrate-hosts. Environmental conditions here can either favor or reduce parasite infection. We discuss the implications of these discrepancies for conservation and ecological studies of infectious diseases in natural populations.

Cryptosporidium spp. infection in captive exotic mammals was investigated using staining and molecular biological methods. A total of 323 fecal samples from 100 mammalian species (62 Artiodactyla, 33 Rodentia, 3 Perissodactyla, and 2 Paenungultata) in 4 zoological gardens in the Czech Republic was examined. Only in a reticulated giraffe (Giraffa camelopardalis reticulata) sample was Cryptosporidium sp. infection detected. The partial small subunit rRNA sequence obtained from the isolate was identical to sequences of Cryptosporidium muris in rock hyrax (Procavia capensis) and Bactrian camel (Camelus bactrianus). Neonatal BALB/c mice inoculated with 1 × 10³ fresh oocysts of the C. muris giraffe isolate did not produce a detectable infection.

For the first time, a palaeoparasitological study was performed on 12 mummies from a Christian cemetery excavated in El-Deir, Oasis of Kharga, Egypt. The analysis revealed the presence of a tapeworm, probably Taenia sp., in a single individual. The presence of just the presumed taeniid egg is surprising and raises the question of the relationship between residents of Egyptian oases and those residing in the Nile Valley. The result suggests information on the health status of the ancient oasis population and re-enforces a hypothesis regarding possible social stratification of the inhabitants. The work must be continued if we are to acquire additional knowledge dealing with life in ancient Egyptian oases.

Plasmodium spp. development within mosquito vectors includes invasive ookinete and sporozoites stages, which display gliding movements during invasion of the midgut and salivary glands, respectively. Sporozoite gliding has been well documented, and several proteins released during sporozoite's locomotion have been described. However, proteins on the ookinete gliding trails are poorly described. In the present study, we documented that 2 proteins, Pbs25 and circumsporozoite thrombospondin-related protein (CTRP), are released during Plasmodium berghei ookinete gliding on a solid surface substrate.

Immunocytochemical examination of onchocercal nodule tissues containing adult Onchocerca volvulus using immuno-markers for blood and lymphatic vessels (vWF, D2-40, podoplanin, Prox-1, and Lyve1) shows a distinct pattern of distribution of these vessels within nodules. Blood vessels were commonly seen associated with organized lymphoid cellular aggregates in the both the outer and inner areas of the nodules. In contrast, the majority of the lymphatic vessel positivity was seen in the central zone in close apposition to the adult parasites, and the remainder usually associated with microfilariae in the outer areas of the nodule. These findings suggest an intimate relationship between adult O. volvulus and lymphatic vessels, including the likely proliferation of lymphatic endothelial cells (lymphangectasia) akin to that seen with other filariae. These findings indicate that adult O. volvulus may migrate via the lymphatic system, and that clinical manifestations of this disease that involve tissue edema may be the result of the location of these worms in the lymphatic system.

Thirty-four amphibians (6 families, 12 species) and 28 reptiles (5 families, 16 species) from Paraguay were examined for helminths. Fifteen (44%) amphibians and 6 (21%) reptiles were found to harbor at least 1 species of helminth; 4 (12%) amphibians and 2 (7%) reptiles harbored multiple infections. Three species of Cestoidea and 17 species of Nematoda were found in the herptiles surveyed. Fourteen new host and 12 new locality records are documented, including the first report of the filaroid nematode, Macdonaldius grassi (Caballero, 1954) Chabaud and Frank, 1961, from South America.

Plasmodium vivax is the second leading cause of malaria worldwide. Invasion of human erythrocytes by P. vivax merozoites is dependent upon the interaction between the parasite Duffy binding protein (PvDBP) and the erythrocyte Duffy antigen receptor. Therefore, disruption of this vital interaction is an attractive target for therapeutic intervention. Although Aotus nancymaae is a commonly used primate model for human P. vivax infections, it has not been confirmed that the interaction between Ao. nancymaae erythrocytes and P. vivax is Duffy antigen dependent. Our results indicate that normal Ao. nancymaae erythrocytes readily bind to PvDBPII and that this binding is completely abolished with chymotrypsin treatment of the erythrocytes. Furthermore, the results of our inhibition assays show a dose-dependent decrease in binding with increasing amounts of anti-PvDBPII polyclonal rabbit sera or anti-Fy6 monoclonal antibody. These data indicate that the interaction between Ao. nancymaae erythrocytes and P. vivax DBPII is Duffy antigen dependent, validating this model system for in vivo studies of anti-PvDBP inhibition.

Canine leishmaniasis and American trypanosomiasis (AT) are caused by related hemoflagellated parasites, Leishmania spp. and Trypanosoma cruzi, which share several common host species. Dogs are reservoirs for human infections by both pathogens. We determined the prevalence of antibodies to Leishmania spp. and T. cruzi in dogs from Grenada, West Indies. We examined 70 dog sera using the qualitative immunochromatographic dipstick tests (ICTs) based on recombinant antigens specific for visceral leishmaniasis and AT. Antibodies to visceral Leishmania were not detected in Grenadian dogs by ICT. Using the canine dipsticks for AT, antibodies to T. cruzi were determined in 3 (4.3%) of the 70 dogs. Results from this study indicate that dogs in Grenada are exposed in low levels to T. cruzi, but not to visceral Leishmania spp. at all.

The global distribution of leishmaniasis is rapidly expanding into new geographic regions. Dogs are the primary reservoir hosts for human visceral leishmaniasis caused by infection with Leishmania infantum. Natural infections with other Leishmania spp. can occur in dogs, but their role as reservoir hosts for other species of Leishmania is uncertain. Leishmania donovani is traditionally considered a visceralizing anthroponotic species; however, cutaneous leishmaniasis caused by L. donovani has been reported in Sri Lanka. In the present study, sera from 114 dogs in Sri Lanka were examined for antibodies to visceralizing Leishmania spp. Sera were tested by the canine immunochromatographic strip assays based on recombinant K39 antigen. Anti-Leishmania spp. antibodies were detectable in 1 of 114 (0.9%) dogs from Sri Lanka. Nonetheless, serological evidence suggests that leishmaniasis may be an emerging zoonosis in Sri Lanka.

We exposed 2 groups of young-of-the-year Arctic charr (Salvelinus alpinus) singly to 54 ± 2 (mean ± SE) Diplostomum spp. cercariae that had emerged from 4 randomly sampled snail hosts (Lymnaea stagnalis). The rearing tanks of the fish received Diplostomum spp. cercariae via the incoming water; therefore, 18 of the 36 fish had parasites in their eyes before the experimental exposure. Of the Diplostomum spp. cercariae presented to the fish, 19% penetrated and 46% of those that had penetrated the fish migrated successfully to the lens of the fish eye. The migration success of Diplostomum spp. from the site of penetration to the fish eye lens was lower in the previously parasitized (37.0 ± 8.7% [mean ± SE] adjusted with the number of penetrated cercariae) than in the unparasitized fish (55.3 ± 8.8%) and differed between the individual snail host from which the cercariae had emerged. In addition, the migration success of Diplostomum spp. decreased with an increase in the number of the cercariae that penetrated the fish. At the individual snail host level, there seemed to be a trade-off between penetration and migration ability of the cercariae. The results indicate that success of Diplostomum spp. in penetration and especially in migration to the fish eye is affected by both the molluscan first intermediate host and the piscine second intermediate host.

Freshwater fish helminth parasites are undoubtedly the most well-known group among vertebrate parasites in Mexico; these parasites include 37 adult and 43 larval (metacercariae) species of trematodes, 62 monogeneans, 15 adult and 18 larval (metacestode) cestodes, 6 adult and 4 larval (cystacanth) acanthocephalans, and 54 adult and 15 larval nematodes (L3). Here, we evaluate the extent of the freshwater fish helminth inventory of Mexico using species accumulation curves and discuss the implications of DNA-based taxonomic methods in our understanding of the helminth biodiversity in freshwater fishes. Future directions for the study of the helminth parasites of freshwater fishes are proposed, particularly the role of DNA-based species delimitation criteria in recognizing parasite species, but also in discovering cryptic species. Species accumulation curves indicate that even though the asymptote has not been reached, a tendency toward stabilization is observed in all taxonomic groups except monogeneans. We suggest, therefore, that the inventory, as conventionally understood, is nearing completion for most groups. We suggest that future survey work aimed at enhancing the biodiversity inventory should be strategic, i.e., it should combine the need to target missing components of the host spectrum with the choice of appropriate drainages based on biogeographic, faunistic, and hydrologic data. We also posit that the future belongs to DNA-based taxonomic approaches that aim to uncover previously unrecognized biodiversity.

Three Haliotrema spp. are reported from the Vietnamese grouper. Morphological and morphometric characters show minor deviations from original descriptions of H. cromileptis Young, 1968 and H. epinepheli Young, 1968. The third encountered species (Haliotrema sp.) appears to be new to science. Genetically, H. cromileptis clusters with Bravohollisia, Pseudohaliotrema, and Haliotrema. The group is well supported by partial large subunit rDNA (LSU), complete small subunit rDNA (SSU), and partial SSU ITS1 rDNA analyses. Ingroup phylogenetic relationships are not well resolved. Haliotrema cromileptis, H. fleti Young, 1968, and Pseudohaliotrema sphincteroporus Yamaguti, 1953 are closely related to a monophyletic group of 5 Haliotrema spp. characterized by bell- or horn-shaped bases of the male copulatory organ (MCO), which contains an accessory piece. Based on SSU rDNA, H. cromileptis is a sister species to P. sphincteroporus and, together, they form a clade to 3 other Haliotrema spp. characterized by a bell-shape based MCO with an accessory piece. Data analysis conducted on partial SSU ITS1 rDNA confirms the close phylogenetic relationship of H. cromileptis, H. fleti, H. chenhsintaoi Zhang, 2001 (possessing a horn-shaped base of the MCO), and Bravohollisia rosetta Lin, 1995. However, because major differences in diagnostic characters exist, this genetic relationship needs further elucidation.

A detailed redescription of Gyropus parvus (Insecta: Phthiraptera: Amblycera: Gyropidae) is given based on specimens collected from the type host, Ctenomys colburni Allen 1903, and the type locality, Estancia Huanuluán, Provincia de Rio Negro, Argentina. We expand and provide new chaetotaxy. New scanning electron microscopy images showing microstructural details of adults and eggs of G. parvus obtained from topotype specimens are included. Sexual dimorphism was mainly shown by differences in body size and abdominal chaetotaxy, with females being 17.5% larger than males and with more setae in each cluster. Significant differences between males and females were also observed in sternal plate measurements. Features described here show homogeneity within type host population. This information contributes to our knowledge of intra- and inter-specific variability for parasite populations. Our investigation constitutes the first collection of G. parvus from the type host and locality since it was described.

With the Eimeria spp. populations that infect chickens used as a model for coccidian biology, we aimed to survey the transcriptome of Eimeria maxima and contrast it to the 2 other Eimeria spp. for which transcriptome data are available, i.e., Eimeria tenella and Eimeria acervulina. The asexual intracellular development stage, the merozoite, was specifically examined, and we used expressed sequence tag (EST) analysis to provide experimental evidence of transcription and a framework for understanding the merozoite stage of E. maxima. Of 2,680 individual ESTs obtained, 48.2% shared most significant (E < 10⁻⁵) homology to sequences from other apicomplexan species, primarily other Eimeria spp. and Toxoplasma gondii, and 47.5% were unique. Annotation of these ESTs enabled categorization to putative biological function and revealed an emphasis on translation, cytoskeleton, metabolism, signaling, transport, and protein folding, as well as the apicomplexan specific surface antigens and micronemes. Comparative analysis of abundantly expressed transcripts from merozoites of the 3 Eimeria spp. revealed a novel transcript common to all 3. Sharing no significant homology to any other sequence in public databases, this transcript was predicted to encode an Eimeria-specific protein (ESP) with 166–178 amino acids and 58.9–65.1% interspecific identity. A predicted signal peptide was identified, consistent with the assumption that ESP is a secreted protein. These annotated ESTs from E. maxima merozoites provide a resource for intra- and interspecific comparative analyses that will be useful in distinguishing the unique biology of coccidian parasites in relation to the diverse phylum of Apicomplexa.

Goblet cell hyperplasia and mucin hypersecretion are important for the expulsion of the intestinal trematode, Gymnophalloides seoi, from mice. However, regulatory mechanisms underlying these processes remain elusive. To better understand the effects of G. seoi antigen on the host's intestinal epithelial cells, we determined whether G. seoi induces expression of Toll-like receptors (TLRs) and mucin-related (MUC) genes on a human intestinal epithelial cell line (HT29 cells). We treated HT29 cells with G. seoi or other adult helminth antigens and measured mRNAs of TLRs and MUCs. We also performed reverse transcriptase-polymerase chain reaction (RT-PCR) and flow cytometry to determine whether TLR and MUC expression is regulated by interferon (IFN)-γ, interleukin-4, or monoclonal antibodies (mAbs) against G. seoi 46 kDa antigen. Gymnophalloides seoi antigen significantly induced expression of TLR2 and MUC2 in HT29 cells, and IFN-γ was found to upregulate TLR2 expression on the surface of the cells. The expression of MUC2 was increased by IFN-γ, but was decreased significantly via the combination of mAbs-to-human TLRs and G. seoi antigen. These results demonstrated that G. seoi antigen upregulates TLR2 and MUC2 expression on human intestinal epithelial cells. These effects reflect a helminth-induced, IFN-γ–dependent, and innate mucosal immune mechanism in this human intestinal cell line.

Although human infections caused by Trichinella sp. have not been reported in Finland for several decades and Trichinella sp. infection in pork has become virtually extinct in the last decade, sylvatic Trichinella spp. infection is still highly prevalent in Finland. Muscle digestion of 2,483 carnivorous wild animals from 9 host species during 1999–2005 showed 617 positive animals (24.8%). Molecular identification from 328 larval isolates revealed 4 different endemic Trichinella species, i.e., T. nativa, T. spiralis, T. britovi, and T. pseudospiralis. Seven percent of the infected animals carried mixed infections. Trichinella nativa was the most common species (74%), but T. spiralis was identified in 12%, T. britovi in 6%, and T. pseudospiralis in 1% of the animals. Host species showed different sample prevalence and Trichinella species distribution. Geographical distribution also varied, with the southern part of the country having significantly higher percentages than the northern part. Infection density was dependent on both the infecting Trichinella species and the host species. Trichinella spiralis was discovered in areas with no known domestic infection cases, indicating that it can also occur in the sylvatic cycle. Raccoon dogs and red foxes are the most important reservoir animals for T. spiralis, as well as for the sylvatic Trichinella species in Finland.

The molecular identification and histopathology are described for the parasitic larvae of a nematode species present in the abdominal cavity of Atlantic salmon (Salmo salar) grilse caught in fish traps on their natal river in the west of Ireland and post-smolts collected during experimental trawls on the continental shelf edge of the northeast Atlantic Ocean. Larvae in the adult and juvenile salmon were identified as Anisakis simplex sensu stricto by PCR amplification and RFLP and sequencing of the ITS gene and PCR amplification and sequencing of the cox2 gene. Parasitic nematode larvae in the grilse were either encapsulated in the abdominal mesentery associated with the pyloric ceca or on the serosal surface of the liver and in the vent region. In some fish, larvae were found in the parenchyma of the liver and muscularis circularis of the intestine. In general, the larvae induced a limited cellular response apart from the occurrence of focal melanin macrophage aggregates and individual eosinophilic granular cells in the connective tissue capsule. Melanin macrophage aggregates were also present among the hepatocytes adjacent to encapsulated larvae in the liver. The reaction to the parasites was more severe in the wall of the intestine. Encapsulated nematode larvae caused displacement, vacuolation, and necrosis of the circular muscle fibers. The stratum compactum was also disrupted with focal areas of degeneration. Overall, the intestinal wall had a hypercellular appearance with extensive cellular infiltration comprising eosinophilic granular cells, macrophages, lymphocytes, and fibrocytes. The post-smolts were caught in May during the early oceanic phase of their life cycle. In these fish, A. simplex sensu stricto larvae were found lying free on the serosal surface of the intestine and liver without any apparent histologic changes. This is the earliest in the marine migration of Atlantic salmon that A. simplex sensu stricto infection has been recorded.

Since the 1970s, no information on simian malaria has been documented in Taiwan, an area that is free from human malaria. To update the prevalence of simian malaria, a molecular-based survey was performed. Blood samples from 286 Formosan macaques (Macaca cyclopis) were tested for Plasmodium species by microscopy and nested polymerase chain reaction. Furthermore, the field isolates were characterized by sequencing the 42-kDa fragment of the merozoite surface protein 1 (MSP-1₄₂). Of the 286 blood samples analyzed, 7 (2.4%) were positive by microscopy and nested PCR. All malaria-infected Formosan macaques were those collected from southern Taiwan, whereas no evidence of malarial parasites was observed among monkeys from eastern and northern Taiwan. Molecular and phylogenetic analyses based on the asexual stage small subunit ribosomal RNA (SSU rRNA) gene clearly identified these samples as a single infection with Plasmodium inui. Furthermore, phylogenetic analysis of the MSP-1₄₂ gene showed that the 7 field isolates were closely related to P. inui strains Taiwan I and II, which were obtained from Formosan macaques in 1963. These findings indicate that P. inui is the only cause of simian malaria in Taiwan, has been circulating in Formosan macaques at least for 46 yr, and has a geographic preference for southern Taiwan.

The prevalence and parasitemia of Hepatozoon caimani in the natural population of the caiman, Caiman yacare, from the Pantanal area, state of Mato Grosso do Sul, central Brazil, were evaluated according to gender and month of capture from July 2006 to February 2008. Blood samples were obtained bimonthly from a total of 229 caimans, and 76% were positive. Prevalence varied significantly according to sampling month and animal size. Almost all adults (100%) and young-adults (97%) were positive, while 63% of juvenile caimans were positive and all of the youngest individuals were negative. These results indicate that caimans are infected for the first time as juveniles. The mean parasitemia in blood was 13.5 ± 13.0 (n  =  174; 1–96 parasites) and did not significantly vary with respect to gender, month of sampling, size, or weight of the caiman. The frequency distribution of parasites in the caiman population was aggregated. Differences in feeding habits and exposure to vectors between the youngest caimans and juveniles are hypothesized as the main risk factors for caimans to acquire H. caimani in central Brazil.

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

Avian coccidiosis, a major parasitic disease of poultry, is caused by Eimeria spp. infection. It inflicts severe economic losses on the poultry industry worldwide. To further understand the molecular basis of sporulation and invasion of Eimeria spp., suppression subtractive hybridization and microarray approaches were combined to identify novel and important genes involved in the development and invasion of the early stages of Eimeria tenella. Three subtractive cDNA libraries were constructed for 3 stages of E. tenella including unsporulated oocysts, sporulated oocysts, and sporozoites. A subset of clones was selected from the 3 subtractive libraries to construct cDNA microarrays. Microarray analysis was used to assay expression changes of these clones. A total of 657 valid expressed sequence tags (ESTs) was obtained, including 119 unique sequences, 31 from sporulated oocysts and 88 from sporozoites. Homology searches of the public sequence databases showed that, among the 119 ESTs, 32 genes encoded proteins homologous with previously reported proteins including microneme proteins and surface antigen proteins of E. tenella, small heat shock proteins, rhoptry proteins of Toxoplasma gondii, and calcium-dependent protein kinase of Plasmodium spp. Thus, the remaining 87 ESTs have not previously been reported. Further characterization of these differentially expressed genes will be useful in understanding those responsible for sporulation, invasion, growth, and development of E. tenella.