The present work deals with the ultrastructure of spermiogenesis and the spermatozoon of Microcotyle pancerii, a gill parasite of meagre Argyrosomus regius collected in Corsican fish farms. Spermiogenesis was rather similar to that observed in other polyopisthocotylean Monogenea. The intercentriolar body was different from that described in digeneans. The nuclear condensation occurred in 2 successive stages. First, during the nuclear migration in the median cytoplasmic process, the nucleus developed a honeycomb-like appearance. Then, after the flagellar fusion, a discontinuous twisting of the chromatin appeared along the nucleus, with this process ending in total nuclear condensation. The structure of the spermatozoon is characterized by 2 axonemes (9 “1” pattern), a single and continuous field of cortical microtubules, a mitochondrion, and a nucleus. Our findings were compared with various ultrastructural features in order to highlight variability within the group.

The epidemiology of Toxoplasma gondii infection in pregnant women in rural Mexico is largely unknown. The sero-epidemiology of T. gondii infection in 439 pregnant women from 9 communities in rural Durango State, Mexico was investigated. Using commercial enzyme-linked immunoassays, sera were tested for T. gondii IgG, IgM, and avidity antibodies. Prevalences of T. gondii IgG antibodies in the communities varied from 0% to 20%. Overall, 36 (8.2%) of the 439 women had IgG T. gondii antibodies. Ten (2.3%) women had also T. gondii IgM antibodies; IgG avidity was high in all IgM-positive women, suggesting chronic infection. None of the women, however, had delivered a known T. gondii-infected child. The seroprevalence was significantly higher (P < 0.05) in women from low socio-economic conditions (14%) than in those with higher socio-economic status (6.6%). Multivariate analysis showed that T. gondii infection was associated with soil floors at home (adjusted OR = 2.89; 95% CI: 1.12–7.49). This is the first epidemiological study of T. gondii infection in pregnant women in rural Mexico.

We examined a population of northern leopard frogs to determine whether sex biases in investment in immunity, previously reported for this host species under controlled exposures to lung nematodes, is predictive of patterns of parasitism in nature. We examined Rhabdias ranae and Haematoloechus spp. infections in 74 breeding adult, 28 non-breeding adult, and 53 juvenile frogs. Contrary to our predictions, R. ranae prevalence and mean abundance were higher in breeding female frogs (prevalence: 39.4%, abundance: 3.05 ± 0.85) than on breeding males (prevalence: 26.0%, abundance: 1.17 ± 0.52), although no sex bias was observed among non-breeding adults or juvenile frogs. Female frogs also carried larger R. ranae worms, on average, than did males (females: 6,407.38 μm ± 153.80; males: 5,198 μm ± 131.09), regardless of age or breeding condition. We observed no sex-linked patterns of parasitism by Haematoloechus spp. worms in either adult or juvenile frogs. Alternative hypotheses, such as differences among sexes in the selection of thermal clines for hibernation, may explain the observed female bias in parasitism by nematode lungworms in nature and, thus, need to be considered.

Host response to parasite exposure is an important determinant of the outcome of host–parasite interactions. Factors such as host body condition and age can strongly influence host response to parasites and infection. This study followed Echinostoma revolutum infection levels in larval green frogs (Rana clamitans) exposed at 2 different ages. Tadpoles at early developmental stages are more susceptible to the adverse effects of echinostomes. Green frog tadpoles approximately 2 wk apart in age and of the same developmental stage can exhibit dramatically different responses to echinostome infection, with the younger tadpoles having high rates of mortality and the older tadpoles experiencing no mortality. The goal of the present study was to begin to explore the mechanism underlying the striking age-dependent response of tadpoles to echinostomes. I conducted 2 controlled infection laboratory experiments in which tadpoles were preserved at 6 time points ranging from 4 hr to 1 wk post-infection (PI). Tadpoles infected at the younger age did not eliminate echinostome metacercariae. However, tadpoles that were 13 days older at the time of echinostome exposure steadily eliminated metacercariae during the hours and days PI. The absence of echinostome cyst elimination in the younger tadpoles likely contributes to their elevated, infection-induced mortality rates.

Eight new species of lice in the genus Columbicola Ewing are described: C. harbisoni (type host: Phaps histrionica (Gould)), C. koopae (type host: Geophaps scripta (Temminck)), C. eowilsoni (type host: Geophaps smithii (Jardine and Selby), C. wombeyi (type host: Geophaps plumifera Gould), C. masoni (type host: Petrophassa rufipennis Collett), C. waiteae (type host: Columba leucomela Temminck), C. rodmani (type host: Geopelia humeralis (Temminck)), and C. smithae (type host: Turtur brehmeri (Hartlaub)). Also, we provide a comprehensive checklist for the 88 known species of Columbicola (Phthiraptera: Ischnocera) and their pigeon and dove hosts (Aves: Columbiformes).

The louse genera Brueelia (Ischnocera) and Myrsidea (Amblycera) are broadly codistributed on songbirds (Passeriformes), but differ in a variety of life history characteristics. We used mitochondrial and nuclear DNA sequences to assess levels of genetic divergence and reconstruct phylogenies of these 2 genera, focusing especially on Catharus thrushes in North America. We then qualitatively compared the phylogenies and levels of divergence within these 2 genera of codistributed parasites. Neither Brueelia nor Myrsidea appears to cospeciate with Catharus thrushes or passerine birds in general. The Myrsidea phylogeny exhibits significant levels of biogeographic structure, whereas the Brueelia phylogeny does not. Myrsidea and Brueelia also differ in their levels of intra-generic genetic divergence, with Myrsidea showing higher levels of genetic divergence and host specificity than Brueelia. Our genetic data support traditional morphology-based taxonomy in several instances in which the same species of Brueelia has been reported on multiple host taxa, e.g., all migrant Catharus spp. carry B. antiqua, with little haplotype divergence. Myrsidea found on each Catharus sp. are in general genetically distinct, except for M. incerta, which parasitizes both Catharus ustulatus and Catharus minimus. The strong biogeographic signal in the Myrsidea phylogeny and higher relative levels of host specificity of Myrsidea spp. suggest that infrequent host-switching, followed by speciation, is shaping the evolutionary history of this group. In contrast, the relatively lower host specificity of Brueelia spp. suggests that host-switching, combined with more frequent ongoing dispersal, has been more important in the evolutionary history of Brueelia.

We studied the deposition of pupae of the winged bat fly Trichobius sp. (caecus group; Diptera), an ectoparasite of Natalus stramineus (Chiroptera, Natalidae), in a natural cave in Tamaulipas, Mexico. For the first time, we show a strong spatial segregation of populations of a streblid bat fly at different stages of development. Using molecular techniques we were able to match developmental stages to adults. Only 5 pupae were present in the main bat roosts. The overwhelming majority occurred exclusively in the bat flyway passages at a considerable distance from roosting bats. Pupal density corresponded positively with the average flight height of bats in the cave passage. Taken together, observations suggest that these ectoparasites must actively seek out their hosts by moving onto passing or roosting bats. The scarceness of pupae in the main roost may be dictated by environmental constraints for their development. The estimated population of viable pupae far exceeds the population of imagoes on the bats, and predation on adults by spiders is common.

Gyrodactylus mexicanus n. sp. and Gyrodactylus lamothei n. sp. are described from the fins and skin of Girardinichthys multiradiatus, an endemic freshwater fish from central Mexico. Gyrodactylus mexicanus is compared to other Gyrodactylus species that parasitize Fundulus spp., the phylogenetically closest group to the Goodeidae from North America. Gyrodactylus mexicanus is distinguished by having large anchors with well-developed superficial roots, enlarged hooks with a proximally disrupted shank (ligament), and a ventral bar with 2 poorly developed anterolateral projections and a small medial process. Gyrodactylus lamothei is distinguished from G. mexicanus and from other species of Gyrodactylus on the North American continent by having anchors with a sclerite on the superficial root and robust hooks with a straight shaft and a recurved point.

Little is known concerning the epidemiology of Toxoplasma gondii infection in people and animals in rural Mexico. Serum samples and tissues from 150 dogs (Canis familaris), 150 cats (Felis catus), 65 opossums (Didelphis virginianus), 249 rats (Rattus spp.), 127 mice (Mus musculus), and 69 squirrels (Spermophilus variegatus) from the Durango area were evaluated for T. gondii infection. Using a modified agglutination test and a serum dilution of 1:25, antibodies to this parasite were found in 68 (45.3%) of 150 dogs, 14 (9.3%) of 150 cats, 11 (16.6%) of 66 opossums, 2 (0.8%) of 249 rats, 4 (3.1%) of 127 mice, and 0 of 69 squirrels. Tissues (brain and heart) of dogs, cats, opossums, rats, mice, and squirrels were bioassayed in mice for the presence of T. gondii. Viable T. gondii was isolated in tissues from 3 of 28 seropositive dogs and 5 of 8 seropositive cats, but not from the other animals. The DNA obtained from the 3 T. gondii isolates from dogs, 6 isolates from 5 cats, and 4 isolates from free-range chickens from Mexico, previously isolated, were genotyped. The PCR-RFLP typing, which used 11 markers (B1, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico), identified 5 genotypes. One genotype (the 4 chicken isolates) belongs to the clonal Type III lineage, three genotypes were reported in previous reports, and 1 genotype is unique.

Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian states, using modified agglutination test (MAT titer ≥1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100% of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans.

A survey was performed between June and September 2006 along the Portuguese coast to assess the prevalence of digenean parasitism in the netted whelk Nassarius reticulatus, a bioindicator of tributyltin (TBT) pollution. It was also intended to evaluate the effect of parasites on the reproduction of this gastropod species and their interference on the development of imposex (the superimposition of male characters onto prosobranch females) and male penis, based on field data. Five digenean species (Lepocreadium album, Gynaecotyla longiintestinata, Himasthla quissetensis, Diphtherostomum brusinae, and Cardiocephalus longicollis), plus 1 unidentified species, were found to infect N. reticulatus. Parasitism was spread along the Portuguese coast, but the higher values of prevalence were found in sheltered inshore areas where up to 67.4% of the animals were affected per sampling station. Parasitism has a castrating impact on the whelks and a reducing effect on male penis size, which causes serious disorders in the reproduction of N. reticulatus and may have an important impact in its population dynamics. However, no relationship between imposex severity and parasite infestation was found.

The protozoan parasite Trypanosoma cruzi is the causative agent of Chagas disease. The chronic stage of infection is characterized by a production of neutralizing antibodies in the vertebrate host. A polyclonal antibody, anti-egressin, has been found to inhibit egress of parasites from the host cell late in the intracellular cycle, after the parasites have transformed from the replicative amastigote into the trypomastigote. It has also been found that BALB/c mouse fibroblasts in the late stages of parasite infection become permeable to molecules as large as antibodies, leading to the possibility that anti-egressin affects the intracellular parasites. This project addresses the fate of the intracellular trypomastigotes that have been inhibited from egressing the host cell. Extended cultures of infected fibroblasts treated with chronic mouse serum reduced parasite egress at all time points measured. Parasites released from infected fibroblasts treated with chronic serum had a reduced ability to infect fibroblasts in culture, yet did not lose infectivity entirely. Absorption of chronic serum with living trypomastigotes removed the anti-egressin effect. The possibility that the target of anti-egressin is a parasite surface component is further indicated by the agglutination of extracellular trypomastigotes by chronic serum. The possibility that cross-linking by antibody occurs intracellularly, thus inhibiting egress, was reinforced by cleaving purified IgG into Fab fragments, which did not inhibit egress when added to infected cultures. From this work, it is proposed that the current, best explanation of the mechanism of egress inhibition by anti-egressin is intracellular agglutination, preventing normal parasite-driven egress.

Temnocephala talicei was described from Uruguayan material found mainly on Aegla prado. Its original description was based principally on its distinctive penial stylet, and, therefore, many aspects of the species anatomy that are now important in the taxonomy of the genus were not contemplated; type specimens were never designated; and the material used to describe the species has been lost. The present work provides a redescription of the species based upon material collected from the type host and in the type locality. Given the possibility of confusion in its identification, and in order to preserve its taxonomic stability, a neotype of T. talicei is designated. The closest species to T. talicei is Temnocephala mertoni, from which it can be separated by the presence of conspicuous intestinal septa, a markedly asymmetrical sphincter in the vagina, markedly lobed testes, a straight penial stylet with a non-sinuous distal portion of the shaft, a characteristic large, ‘mouthpiece’-shaped introvert without discrete thickenings, and approximately 10 crowns of well-developed spines of decreasing length placed throughout the whole extension of the introvert. A comparison of material from Uruguay, Argentina, and Paraguay shows that there is little variation in the morphometry of the species. Finally, a comparison of the post-tentacular syncytia of T. talicei and T. mertoni shows that this is not always a species-specific character.

Forty-four splenectomized Aotus nancymaae monkeys were infected with 6 different strains of Plasmodium cynomolgi, 11 via trophozoites and 33 via sporozoites. Sporozoites from Anopheles dirus, Anopheles freeborni, Anopheles gambiae, Anopheles maculatus, and Anopheles stephensi resulted in prepatent periods ranging from 9 to 39 days (median of 15 days). Importantly, relapse was demonstrated in 5 of 5 sporozoite-induced infections with the Rossan strain following treatment with chloroquine.

The cercaria of Bucephalus minimus infects the digestive gland and gonads of its first intermediate host, the edible cockle, Cerastoderma edule. Light microscopy (LM) and scanning electron microscopy (SEM) of the cercaria showed a tail formed by a central stem, with 2 long contractile arms presenting distinct morphological surfaces. The encysted metacercaria naturally infected the flathead grey mullet, Mugil cephalus. The cysts found in the heart, liver, and spleen were shown to be identical by the internal transcribed spacer (ITS1) sequence and morphological features and were associated with encapsulation, recruitment of cell infiltrates, and presence of melanomacrophages and adipose tissue. To establish the life cycle, we compared the ITS1 sequence in an adult from the known definitive host, Dicentrarchus labrax; encysted metacercariae from the liver, heart, and spleen of M. cephalus; and a cercaria from C. edule. With this comparison, we determined that they had a 100% similarity. Therefore, the ITS1 sequence data clearly indicate that these 3 parasitic stages belong to the same species, i.e., B. minimus.

We present the first demonstration of oral transmission of Trypanosoma cruzi to raccoons (Procyon lotor), a natural reservoir host in the United States, by ingestion of trypomastigotes and infected bugs, but not infected tissue. To investigate an alternative, non-vector–based transmission method, we tested the hypothesis that raccoons scavenging on infected hosts results in patent infection. Macerated tissue from selected organs infected with amastigote stages of T. cruzi was orally administered to experimental groups of raccoons (n = 2/group) at 2, 12, or 24 hr after collection of the tissue samples. Additionally, raccoons (n = 1) in control groups were inoculated intravenously or per os with trypomastigotes. To further elucidate transmission routes of T. cruzi to raccoons, infected Rhodnius prolixus were fed to raccoons (n = 2). Raccoons did not become infected after ingestion of amastigote-infected tissues as evidenced by negative polymerase chain reaction results from blood and tissue, lack of seroconversion, and negative parasitemias. However, per os transmission can occur by ingestion of the infective trypomastigote stage or infected reduviid bugs. We conclude from these findings that oral transmission of T. cruzi may be a route of infection for wildlife in sylvatic cycles, but the scavenging behavior of animals is not likely a significant transmission route.

Paleoparasitological research has made important contributions to our understanding of parasite evolution and ecology since the 1960s. Since then, most studies have focused on paleoparasitological evidence from single sites or samples. With the development of high throughput sequencing techniques, new avenues of investigation for paleoparasitological material are opening up. Here, I provide an overview of recent developments and highlight how these results will broaden the scope of the field, placing paleoparasitology at the interface of a wide array of studies, including parasitology, climate change, human evolution, and evolutionary processes.

According to current knowledge, sexual development of the apicomplexan parasite Neospora caninum takes place in the canine intestine. However, to date there is no information on the interaction between the parasite and the canine intestinal epithelium, and, next to the clinical and in vivo research tools, an in vitro model comprised of canine intestinal cells infected with N. caninum would be very helpful for investigations at the cellular level. Following the isolation of cells of neonatal canine duodenum and growth of cell cultures to monolayers for 5–6 days, canine intestinal epithelial cells were exposed to cell culture– derived N. caninum tachyzoites and bradyzoites. The host cells remained viable during in vitro culture for an average of 2 wk. During this time span, N. caninum was found to readily adhere to any surface area of these cells, but infection took mostly place at sites where microvilli-like structures were missing, e.g., at the cell periphery, with tachyzoites exhibiting at least 3–4 times increased invasive capacities compared to bradyzoites. Once intracellular, parasites resided within a parasitophorous vacuole, moved toward the vicinity of the nucleus and the more distal portion of the epithelial cells, and proliferated to form vacuoles of not more than 2–4 parasites, which were surrounded by numerous mitochondria. Immunofluorescence staining and TEM of infected cells showed that the expression of cytokeratins and the structural integrity of desmosomes and tight junctions were not notably altered during infection. Furthermore, no changes could be detected in the alkaline phosphatase activities in cell culture supernatants of infected and noninfected cells. Canine duodenal epithelial cell cultures represent a useful tool for future studies on the characteristics of the intestinal phases of N. caninum infection.

The blood meals of field-collected female Culex territans (Diptera: Culicidae) were concurrently assayed for the presence of trypanosomes and for vertebrate host identification. We amplified vertebrate DNA in 42 of 119 females and made positive identification to the host species level in 29 of those samples. Of the 119 field-collected Cx. territans females, 24 were infected with trypanosomes. Phylogenetic analysis placed the trypanosomes in the amphibian portion of the aquatic clade of the Trypanosomatidae. These trypanosomes were isolated from Cx. territans females that had fed on the frog species Rana clamitans, R. catesbeiana, R. virgatipes, and Rana spp. Results support a potential new lineage of dipteran-transmitted amphibian trypanosomes may occur within the aquatic clade. The frequency in which female Cx. territans acquire trypanosomes, through diverse feeding habits, indicates a new relationship between amphibian trypanosomes and mosquitoes that has not been examined previously. Combining Trypanosoma species, invertebrate, and vertebrate hosts to existing phylogenies can elucidate trypanosome and host relationships.

A new species of parasitic nematode, Capillostrongyloides congiopodi n. sp. (Capillariidae), is described based on specimens collected from the gall bladder of the horsefish, Congiopodus peruvianus Cuvier and Velenciennes (Congiopodidae, Scorpaeniformes), from the Patagonian Shelf, Argentina (45–48°S; 60–64°W). Among the 9 species described so far in the genus, the new species most closely resembles C. norvegica Moravec and Karlsbakk, 2000, by the presence of its conspicuously elevated anterior vulvar lip in females; however, it is readily distinguished from it by having a larger body size, larger eggs with protruding polar plugs, the shape and length of the spicule, and mainly by the general morphology of the caudal bursa of males. In addition, the site of infection, i.e., stomach versus gall bladder. This is the first Capillostrongyloides species reported from fishes in the southern Atlantic Ocean.

A new nematode species, Freitascapillaria moraveci n. sp., is described; it was obtained from specimens recovered from the gall bladder of the 2-spot livebearer Heterandría bimaculata (Heckel, 1848) from La Antigua River, State of Veracruz, Mexico. The new species is assigned to Freitascapillaria Moravec 1982; it is largely characterized by the posterior end of the males, which is laterally expanded without distinct projections. Freitascapillaria moraveci n. sp. differs from the only other species of the genus, F. maxillosa, by the absence of wing-like cells at the esophago-intestinal junction, which are present in the latter species; stichosoma consists of 30–36 stichocytes (F. moraveci) versus 40–60 stichocytes (F. maxillosa) in both males and females and the presence of a well-developed spicule.

Inglisera cirrohamata (Linstow, 1888) (Nematoda: Acuariidae) is redescribed based on specimens collected from the type host, Phalacrocorax verrucosus (Aves: Phalacrocoracidae), and the type locality, Kerguelen Island, in the southern Indian Ocean. Three new hosts and 2 new localities were recorded, i.e., Phalacrocorax [atriceps] albiventer and P. brasilianus from Puerto Madryn, Argentina and P. magellanicus from Puerto Williams, Chile. The structure of the cephalic ornamentations was observed and fully described. The presence of 6 pairs of postcloacal papillae is confirmed, a description of the tip of left spicule is given, and the presence of post-deirids is reported for the first time. This constitutes the first record of this nematode in South America and provides the first study of the genus by scanning electron microscopy.

A new nematode species, Philometra poblana n. sp., is described based on specimens recovered from skin at the base of the pectoral fins of the cichlid Cichlasoma istlanum (Jordan and Snyder, 1899) from the water spring El Borbollon, in the State of Puebla, Mexico. The new species most closely resembles Philometra gymnosardae and Philometra ophisterni; however, P. poblana can be easily differentiated from the other species by the length of gravid females (7.10–10.43 vs. 14.8– 27.0 and 28.67–39.30 mm, respectively), length of caudal projections (0.015–0.023 vs. 0.047 and 0.006–0.009 mm high, respectively), site of infection (skin at base of pectoral fins vs. abdominal cavity, both species), and the host species (Cichlidae vs. Synbranchidae and Scombridae).

Procamallanus (Spirocamallanus) belenensis n. sp. is described using light and scanning electron microcopy, based on specimens taken from Ageneiosus ucayalensis, Castelnau, 1855 (Pisces: Siluriforme), a freshwater fish commonly called mandubé, in Pará State, Brazil; Procamallanus (Spirocamallanus) rarus is recorded from the same host. To our knowledge there are no previous reports of nematodes from this fish species. Procamallanus (S.) belenensis is characterized by the presence of 8–13 cuticular spiral thickenings coating 2/3 of posterior region of the buccal capsule of males and females (the anterior third is smooth); small sub-equal spicules; 3 and 4–5 pairs of pre- and post-cloacal papillae, respectively; the proportion of muscular and glandular esophagus is 1:1.5–2; males present 2 spike-like projections at the tail tip, which are absent in females by light microscopy. The most closely related species are Procamallanus (Spirocamallanus) inopinatus and Procamallanus (Spirocamallanus) pintoi. The new species differs from the most closely related species by the presence of 8–10 cuticular spiral thickenings in the buccal capsule of males and 9–13 in the females, 8 pairs of male clocal papillae (3 pre- and 5 post-cloacal, respectively), and only males bearing 2 minute, spike-like projections at the posterior end.

A new species of nematode, Cucullanus costaricensis n. sp., is described from the Red Sea catfish Bagre pinnimaculatus from Río Tempisque, Costa Rica. The new species is characterized by the following traits: spicules and gubernaculum length (0.48 ± 0.02 [0.45–0.50] and 0.20 ± 0.02 [0.18–0.21], respectively); a protruding cloacal region in males, and the arrangement of caudal papillae (1 medial unpaired precloacal papilla and 10 pairs of papillae: 3 precloacals [first pair subventral and anterior to ventral sucker; second and third pairs posterior to ventral sucker; second pair asymmetric and subventral; third pair slightly subventral and closer to cloacal opening], 3 subventral adcloacal pairs [pairs 4–6], and 4 pairs of postcloacal papillae [seventh pair lateral, eighth pair subdorsal, ninth and tenth pairs subventral]); and lateral phasmids between the seventh and eighth pairs of postcloacal papillae. This finding represents the first record for a species of Cucullanus in Central America.

New Guinea is one of the most biodiverse regions of the world, particularly in terms of the herpetofauna present, yet surprisingly little is known about the parasites that infect these organisms. A survey of diverse scinid and agamid lizard hosts from this country showed a diversity of malaria parasites infecting these hosts. We combined morphological and morphometric observations of the parasites (primarily gametocytes) along with DNA sequence data from the mitochondrial cytochrome b and cytochrome oxidase I genes and here describe 4 new species of Plasmodium, i.e. Plasmodium minuoviride n. sp., Plasmodium koreafense n. sp., Plasmodium megalotrypa n. sp., and Plasmodium gemini n. sp. A fifth species, Plasmodium lacertiliae Thompson and Hart 1946, is redescribed based on new observations of hosts and localities and additional molecular data. This combined morphological and molecular approach is advised for all future descriptions of new malaria parasite species, particularly in light of situations where every life-history stage is not available.

Ticks, representing 3 species of Amblyomma, were collected from the water python (Liasis fuscus) and 3 additional reptile species in the Northern Territory, Australia, and tested for the presence of Hepatozoon sp., the most common blood parasites of snakes. In addition, blood smears were collected from 5 reptiles, including the water python, and examined for the presence of the parasite. Hepatozoon sp. DNA was detected in all tick and reptile species, with 57.7% of tick samples (n = 187) and 35.6% of blood smears (n = 35) showing evidence of infection. Phylogenetic analysis of the 18S rRNA gene demonstrated that half of the sequences obtained from positive tick samples matched closest with a Hepatozoon species previously identified in the water python population. The remaining sequences were found to be more closely related to mammalian and amphibian Hepatozoon species. This study confirms that species of Amblyomma harbor DNA of the same Hepatozoon species detected in the water pythons. The detection of an additional genotype suggests the ticks may be exposed to 2 Hepatozoon species, providing further opportunity to study multiple host–vector–parasite relationships.

The tapeworm Proteocephalus glanduligerus (Janicki, 1928) Fuhrmann, 1933 (Cestoda: Proteocephalidea), a parasite of clariid catfishes (Clarias spp.) in Africa, is redescribed on the basis of an evaluation of voucher material and newly collected specimens from Clarias gariepinus and C. cf. anguillaris from the Sudan, Ethiopia, and South Africa. The most typical characteristic of P. glanduligerus (previously misspelled as P. glanduligera or P. glanduliger) is the presence of an extremely large glandular apical organ, the size of which is 1.6–3.5 times larger than that of the suckers. In addition, other morphological characteristics unreported in previous accounts are provided, such as an unusual position of osmoregulatory canals (situated close to each other, with the dorsal canal latero-ventral to testes), the presence of a vaginal sphincter, several (usually 3–5) uterine pores, and eggs with paired lateral auricular swellings (extensions) of the outer envelope.

The purpose of this study was to compare the sensitivity and specificity of an ELISA test to detect Fasciola gigantica antigens (coproantigens) in bovine feces, with fecal egg counting and an ELISA for detecting anti–F. gigantica antibodies in serum. Monoclonal antibodies to cathepsin L were generated and used to capture this antigen in feces of infected cattle. Blood, feces, and livers were collected from 150 cattle at an abattoir in Jakarta, Indonesia, for anti-Fasciola antibodies, coproantigen detection, and F. gigantica egg and worm counts. Fluke recovery varied from 1 to 426 per host, with a mean of 32 flukes. The results showed that the sensitivity and specificity of coproantigen detecting ELISA (95 and 91%, respectively) was better than the anti–F. gigantica antibody ELISA (91 and 88%, respectively) and to fecal egg counting (87 and 100%, respectively). The coproantigen ELISA was able to detect 100% of the cattle with >15 flukes. A survey of 305 cattle in central Java over a 10-mo period validated this test in the field, demonstrating a high prevalence of fascioliasis and establishing the test as a useful diagnostic method to determine patent F. gigantica infections in cattle.

The leishmanicidal activity of 15 extracts and 4 pure metabolites obtained from Urechites andrieuxii, Colubrina greggii, Dorstenia contrajerva, and Tridax procumbens was evaluated using the newly developed MTS ({3-(4,5-dimethylthiazol- 2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay, optimized for promastigotes of Leishmania major, Leishmania tropica, and Leishmania aethiopica, as well as for L. aethiopica axenic amastigotes. The assay was then used for calculating the percentage of viable stationary phase parasites after a 24-hr treatment with each plant extract or pure metabolite. The 3 most active samples, 2 from C. greggii (NCG-5C and DCG-3A) and 1 from T. procumbens (TPZ-2A), showed LD₅₀ values of 62.4, 7.2, and 18.5 μg/ml, respectively, on stationary promastigotes, and of 94.2, 27.1, and 95.2 μg/ml, on amastigotes of L. aethiopica. Moreover, TPZ-2A and DCG-3A significantly reduced the percentage of infected monocyte-derived macrophages (THP-1). The percentage of infected cells decreased from 69.9% ± 2.5% to 20.8% ± 2% when the cells were treated with the DCG-3A fraction and to 14.9% ± 0.5% when treated with TPZ-2A, without significantly decreasing the number of human cells. These findings indicate the presence of potentially bioactive metabolites in the roots of C. greggii and in T. procumbens and reflect the importance of pursuing the bioassay-guided purification of these metabolites.

Chagas disease, caused by Trypanosoma cruzi, is a widespread infection in Latin America. Currently, only 2 partially effective and highly toxic drugs, i.e., benznidazole and nifurtimox, are available for the treatment of this disease, and several efforts are underway in the search for better chemotherapeutic agents. Here, we have determined the trypanocidal activity of 2,3-diphenyl-1,4-naphthoquinone (DPNQ), a novel quinone derivative. In vitro, DPNQ was highly cytotoxic at a low, micromolar concentration (LD₅₀ = 2.5 μM) against epimastigote, cell-derived trypomastigote, and intracellular amastigote forms of T. cruzi, but not against mammalian cells (LD₅₀ = 130 μM). In vivo studies on the murine model of Chagas disease revealed that DPNQ-treated animals (3 doses of 10 mg/kg/day) showed a significant delay in parasitemia peak and higher (up to 60%) survival rate 70 days post-infection, when compared with the control group (infected, untreated). We also observed a 2-fold decrease in parasitemia between the control group (infected, untreated) and the treated group (infected, treated). No apparent drug toxicity effects were noticed in the control group (uninfected, treated). In addition, we determined that DPNQ is the first competitive inhibitor of T. cruzi lipoamide dehydrogenase (TcLipDH) thus far described. Our results indicate that DPNQ is a promising chemotherapeutic agent against T. cruzi.

Pectoral muscles from a captive keel-billed toucan (Ramphastos sulfuratus) from Costa Rica were fed to a Toxoplasma gondii– free cat, and the cat shed oocysts. Laboratory mice fed these oocysts developed antibodies to T. gondii in their sera and T. gondii tissue cysts in their brains. The DNA extracted from the brains of infected mice was characterized using 10 polymerase chain reaction–restricted fragment length polymorphic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The isolate designated TgRsCr1 was found to be non-clonal with Type I, II, and III alleles at different loci. This is the first isolation of T. gondii from this host.

The irregular presence and low abundance of wild triatomines inside domiciles make their detection more difficult than that of domiciled species, so that vector surveillance and evaluation of Chagas disease transmission risk are more challenging. We compared timed manual searches, considered as the gold standard, with community-based collections, for their efficacy at monitoring domestic and peri-domestic infestation by non-domiciliated Triatoma dimidiata, and community-based collection was the most sensitive and cost effective. Scaling up community participation permitted investigation of fine temporal variations in infestation by T. dimidiata in over 700 houses. We confirmed a large seasonal infestation during March–July, but weekly and daily collections showed a rather stochastic pattern of bug presence in the houses, even during this period. These data are of key importance for the successful implementation of vector control, and community participation is a method of choice for sustained monitoring of infestation by non-domesticated triatomines.

As shown previously, the nested PCR method of detection of avian blood parasites, commonly referred to as Waldenström's method, sometimes amplifies only 1 parasite species of the several that may be present in the same bird, and not always the one with the highest parasitemia. This result raises questions regarding the use of the molecular method for the identification of bird parasites. Additionally, it is unclear whether the amplified parasite, among the several present in the same host, reflects the intensity of infection. However, I performed statistical analyses on a dataset in which there were multiply infected birds and showed that the parasites with the highest parasitemia are the most likely to be amplified. Such a positive correlation between the likelihood of amplification and the parasitemias of the different blood parasites supports the use of Waldenström's method for the comparison of the parasite content of groups of birds exposed to the same parasites.

Besnoitia besnoiti was isolated from a skin biopsy of a chronically infected cow from central Spain. Zoites released from macroscopic cysts were adapted to its culture in vitro on a MARC-145 cell monolayer. Tachyzoites produced in vitro were either cryopreserved or used for genomic DNA isolation. A 2206 nt sequence containing 18S ribosomal RNA gene, internal transcribed spacer 1 (ITS1), and a partial sequence of 5.8S ribosomal RNA gene was amplified by PCR and sequenced. This sequence showed a 99–100% identity to 18S, ITS1, and 5.8S sequences of B. besnoiti published in databases. After analysis by transmission and scanning electron microscopy of isolated bradyzoites and tachyzoites, it was observed that their ultrastructural morphology coincided with B. besnoiti. The isolate characterized in this study was identified as B. besnoiti on the basis of the disease produced, molecular characteristics, and morphology. The B. besnoiti isolate was denoted as BbSpain-1; it is the first isolate obtained and characterized in Spain and one of the first European isolates adapted to grow in vitro. The isolation and in vitro production of this B. besnoiti isolate offers a good opportunity to study general aspects of bovine besnoitiosis, including epidemiology, pathogenesis, and diagnosis of this re-emergent disease.

In aquatic ecosystems, dense populations of snails can shed millions of digenean trematode cercariae every day. These short-lived, free-living larvae are rich in energy and present a potential resource for consumers. We investigated whether estuarine fishes eat cercariae shed by trematodes of the estuarine snail Cerithidea californica. In aquaria we presented cercariae from 10 native trematode species to 6 species of native estuarine fishes. Many of these fishes readily engorged on cercariae. To determine if fishes ate cercariae in the field, we collected the most common fish species, Fundulus parvipinnis (California killifish), from shallow water on rising tides when snails shed cercariae. Of 61 killifish, 3 had recognizable cercariae in their gut. Because cercariae are common in this estuary, they could be frequent sources of energy for small fishes. In turn, predation on cercariae by fishes (and other predators) could also reduce the transmission success of trematodes.

Analysis of rodent models of inflammatory bowel disease, airways hyper-reactivity, diabetes, and multiple sclerosis has shown that infection with helminth parasites can significantly reduce the severity of the disease. Here, we assessed whether rats infected with the tapeworm Hymenolepis diminuta were protected from gastric ulceration induced by the serosal application of acetic acid. All rats gavaged with infective cysticercoids harbored adult worms when assessed 6 wk later, and acetic acid evoked the expected gastric ulceration. However, infection with H. diminuta did not affect the degree of gastric ulceration at either 3 or 7 days post-acetic acid application, as gauged by ulcer area or histopathology. While the data do not dismiss the possibility that infection with other helminths could be anti-ulcerogenic, they illustrate that ‘helminth therapy’ for inflammatory disease is likely to be both disease- and helminth-specific.

In the summer of 2007, American eels, Anguilla rostrata, from 2 localities on Cape Breton Island, were found to be infected with the swim bladder nematode Anguillicoloides crassus. This is the first documented report of this highly invasive parasite in Canadian waters. More than half of the yellow eels in Mira River (6 of 10), and 1 eel (of 5) from Sydney Harbour were infected. Parasite intensity ranged from 1 to 11 worms per eel. The occurrence of A. crassus at these 2 localities suggests the need for a more extensive survey on the distribution of this exotic parasite in eel populations throughout Cape Breton Island.

The development of microsatellite markers for parasitic nematodes has been hampered by technical difficulties in isolation and PCR amplification. We have investigated the potential for circumventing these problems using microsatellites from 3 trichostrongyloid species on a panel of 7 species. Ten of the 22 PCR primer pairs tested amplified in species other than the target species, usually in closely related species, and 2 new variable loci were discovered in the sheep parasite Trichostrongylus vitrinus. This study provides evidence that cross-species testing of microsatellite primers can be an effective alternative to isolation de novo.

Permanent and well-supported museum or natural history collections provide a solid foundation for the process of systematics research through creation of an empirical record which validates our understanding of the biosphere. We explore the role of museums in ongoing studies of the complex helminth fauna characteristic of pikas (Ochotona spp.) in the American west. These studies address the taxonomy for pinworms of the Labiostomatinae and the problems associated with the absence of adequate type series and vouchers and with misidentifications in original descriptions. We demonstrate that the types for Labiostomum (Labiostomum) coloradensis are identical to some specimens in the syntype series representing L. (Eugenuris) utahensis, although the published descriptions are in disagreement. Both are identical to L. (Eugenuris) talkeetnaeuris and, as a consequence, are reduced as junior synonyms. Only 2 species of large pinworms, namely L. (Labiostomum) rauschi and L. (Eugenuris) talkeetnaeuris, are widely distributed in Ochotona collaris and O. princeps. Although this serves to clarify the taxonomy for species in these genera, prior records remain confused, as representative voucher specimens from all major surveys in North America were never submitted to museum collections. We strongly suggest that type and voucher series should not be held in private or personal collections, where such are eventually lost, discarded, or destroyed through neglect due to inattention and the absence of curation. The potential to accumulate meaningful baselines for assessment of environmental change is jeopardized if materials from survey and inventory are not routinely submitted to museum collections. The capacity of museum repositories, as a focus for systematics, ecology, and evolutionary studies and for the development of resources for biodiversity informatics, continues to be undervalued and poorly utilized by a cadre of scientists who are dependant on accurate and definitive information that transcends specific disciplines.