Crowding, i.e., the size of the infrapopulation inhabiting an individual host, is a major component of parasites' environment, which often influences both morphological and life-history characters (the so-called density-dependent characters) in different parasite taxa. Although crowding equals intensity in case of a single parasite individual, mean intensity of the host population does not define mean crowding of the parasite population. Crowding indices are notoriously hard to handle statistically because of the inherently large number of nonindependent values in data. In this study, we aim to investigate the apparently paradox features of crowding indices and to make some proposals and also to introduce statistical methods to calculate confidence intervals and 1-sample and 2-sample tests for mean crowding. All methods described in this study are supported by the freely distributed statistical software Quantitative Parasitology.

The phylogenetic relationships and historical biogeography of species of Sciadicleithrum parasitizing Neotropical cichlid fishes were studied. Hypotheses were: Did the presence of Sciadicleithrum species in southeast Mexico (SM) and Central America (CA) result from early dispersal of cichlid species from South America (SA) to the north via the Antilles during the Tertiary (24 million years ago [mya]), or did it occur after emergence of the Panama Isthmus in the Pleistocene (2–5 mya)? The Sciadicleithrum phylogeny was based on 19 morphological transformation series, with species of Gussevia and Cichlidogyrus as outgroups. The most parsimonious cladogram had a 40% consistency index, with one clade including all the species of Sciadicleithrum from SA and all but one of those from CA and another with all the Sciadicleithrum species from SM and S. maculicaudae from CA. Results support the late dispersal hypothesis following emergence of the Panama Isthmus. Fifteen species of Sciadicleithrum parasitize 13 cichlid species from SA and CA. In contrast, only 4 species of Sciadicleithrum infect 14 cichlid species from SM. Parasite speciation appears to lag behind host speciation, with 2 equally possible explanations: loss of parasite species and host switching.

Sequence analysis of the ribosomal DNA second internal transcribed spacer (ITS 2) region in 2 spatially distinct populations of Amblyomma americanum (L.) revealed intraspecific variation. Nucleotide sequences from multiple DNA extractions and several polymerase chain reaction amplifications of eggs from mixed-parentage samples from both populations of ticks revealed that 12 of 1,145 (1.0%) sites varied. Three of the 12 sites of variation were distinct between the 2 A. americanum populations, which corresponded to a rate of 0.26%. Phylogenetic analysis based on ITS 2 sequences provided strong support (i.e., bootstrap value of 80%) that wild A. americanum clustered into a distinguishable group separate from those derived from colony ticks.

The majority of patients infected with lymphatic filariae are microfilaremic but tend to manifest little obvious pathology because of the infections. Data collected from the Mongolian gerbil–Brugia spp. model for human lymphatic filariasis suggest this experimental animal model system most closely represents this patient group and will be useful in studying immunological parameters associated with chronic infections. This article reports the quantitation of interleukin (IL)-4, IL-5, IL-10, IL-13, and interferon (IFN)-γ messenger RNA (mRNA) in gerbils after a primary subcutaneous infection with Brugia pahangi. Chronically infected gerbils showed elevated IL-4 in all tissues, compared with earlier time points, linking this Th2 cytokine to the downregulation of responsiveness, which develops in gerbils and humans. Both IL-5 and IL-13 mRNA expression were transient in all tissues. The peak in IL-5 at 14–28 days postinfection reflects the peak of peripheral eosinophilia observed in B. pahangi–infected gerbils. Little IFN-γ mRNA was reported from chronically infected gerbils. The data collected thus far suggest that the expression profile of many of the measured cytokines in B. pahangi–infected gerbils reflects what is seen in an important subset of humans infected with lymphatic filariae, the microfilaremic, asymptomatic patient.

In this study, we evaluate the effect of phospholipid on the adjuvanicity and protective efficacy of liposome vaccine carriers against visceral leishmaniasis (VL) in a hamster model. Liposomes prepared with distearyol derivative of l-α-phosphatidyl choline (DSPC) having liquid crystalline transition temperature (Tc) 54 C were as efficient as dipalmitoyl (DPPC) (Tc 41 C) and dimyristoyl (DMPC) (Tc 23 C) derivatives in their ability to entrap Leishmania donovani membrane antigens (LAg) and to potentiate strong antigen-specific antibody responses. However, whereas LAg in DPPC and DMPC liposomes stimulated inconsistent delayed type hypersensitivity (DTH) responses, strong DTH was observed with LAg in DSPC liposomes. The heightened adjuvant activity of DSPC liposomes corresponded with 95% protection, with almost no protectivity with LAg in DPPC and DMPC liposomes, 4 mo after challenge with L. donovani. These data demonstrate the superiority of DSPC liposomes for formulation of L. donovani vaccine. In addition, they demonstrate a correlation of humoral and cell-mediated immunity with protection against VL in hamsters.

The production of reactive oxygen species by hemocytes from the gastropod Biomphalaria glabrata has been linked to their ability to kill the trematode parasite Schistosoma mansoni. For 2 laboratory strains of the snail, 1 resistant (13-16-R1) and 1 susceptible (MO) to the PR1 strain of S. mansoni, we compared hemocyte production of extracellular hydrogen peroxide when stimulated with the protein kinase C agonist phorbol myristate acetate (PMA). The time course of the PMA-induced response is similar in both strains with respect to onset, peak production, and termination of the respiratory burst. However, the magnitude of the response differs between strains, in that hemocytes from resistant snails generate significantly more hydrogen peroxide. These findings suggest that the capacity to produce hydrogen peroxide could be critical in determining susceptibility or resistance to S. mansoni.

Infections that cause the Gombak and Smithsonian strains of Plasmodium cynomolgi were induced in Macaca mulatta, Aotus lemurinus griseimembra, Aotus nancymai, and Saimiri boliviensis monkeys. Transmission of the Gombak strain to Aotus spp. monkeys was obtained by the injection of sporozoites dissected from the salivary glands of experimentally infected Anopheles dirus and by the bites of infected An. dirus and Anopheles farauti mosquitoes. Two S. boliviensis monkeys were infected via the injection of sporozoites dissected from An. dirus. Prepatent periods in New World monkeys ranged from 14 to 44 days, with a median of 18 days. The Smithsonian strain was transmitted via sporozoites to 1 A. lemurinus griseimembra and 9 A. nancymai monkeys. Prepatent periods ranged from 12 to 31 days.

We describe here a fatty acid–binding protein (FABP) isolated and purified from the parasitic protozoon Giardia lamblia. The protein has a molecular mass of 8 kDa and an isoelectric point of 4.96. A Scatchard analysis of the data at equilibrium revealed a dissociation constant of 3.12 × 10⁻⁸ M when the labeled oleic acid was displaced by a 10-fold greater concentration of unlabeled oleic acid. Testosterone, sodium desoxycholate, taurocholate, metronidazol, and α-tocopherol, together with butyric, arachidonic, palmitic, retinoic, and glycocholic acids, were also bound to the protein. Assays with polyclonal antibodies revealed that the protein is located in the ventral disk and also appears in the dorsal membrane, the cytoplasm, and in the vicinity of the lipid vacuoles.

Superoxide dismutases (SODs; EC 1.15.1.1) play important roles in the protection of the parasites against cellular oxygen–mediated killing of the hosts. A copper/zinc-containing SOD (Cu/Zn-SOD) was identified previously from lung fluke, Paragonimus westermani. To expand our understanding of P. westermani SOD, we isolated a complementary DNA encoding a Cu/Zn-SOD, expressed the active enzyme in Escherichia coli, and characterized its biochemical properties. The deduced amino acid (aa) sequence of the gene shared up to 73.7% identities with Cu/Zn-SODs of other helminths and shared well-conserved characteristic motifs and essential aa residues involved in coordinating copper and zinc enzymatic functions. Recombinant Cu/ Zn-SOD exhibited comparable biochemical properties with that of the native enzyme, including pH optima and potassium cyanide–and hydrogen peroxide–sensitive inhibition profiles. The active enzyme consisted of 2 identical subunits covalently linked by disulfide bonds. The enzyme was constitutively expressed throughout various developmental stages of the parasite. The levels increased as P. westermani matured and plateaued in adult stage. Our result suggests the enzyme might play an important role for parasites to survive in the hosts through its superoxide anion–detoxifying function.

Heat shock protein 90 (Hsp90) is 1 of the most abundant and evolutionarily conserved proteins. In most species, Hsp90 is essential for proper cell function. In this study, we present the molecular analysis of Hsp90 from Eimeria species, the causative agents of avian coccidiosis. The full-length Eimeria acervulina Hsp90 complementary DNA was isolated from intestinal intraepithelial lymphocytes of Eimeria-infected chickens. From evolutionary analysis and sequence identity, it is likely that Eimeria Hsp90 sequences described thus far encode the cytosolic versions of the protein. Although at the nucleotide and amino acid levels Eimeria tenella and E. acervulina Hsp90 are highly similar, their expression profiles differ considerably. Although E. tenella transcripts were detected in all developmental stages tested, E. acervulina transcripts were not found in oocysts undergoing sporulation or in fully sporulated oocysts, suggesting that messenger RNA expression may be regulated quite differently between Eimeria species.

Organotypic slice culture explants of rat cortical tissue infected with Toxoplasma gondii tachyzoites were applied as an in vitro model to investigate host–pathogen interactions in cerebral toxoplasmosis. The kinetics of parasite proliferation and the effects of interferon-γ (IFN-γ) and tumor necrosis factor–α (TNF-α) in infected organotypic cultures were monitored by light microscopy, transmission electron microscopy (TEM), and quantitative polymerase chain reaction (PCR) assay. As assessed by the loss of the structural integrity of the glial fibrillary acidic protein–intermediate filament network, tachyzoites infected and proliferated mainly within astrocytes, whereas neurons and microglia remained largely unaffected. Toxoplasma gondii proliferation was severely inhibited by IFN-γ. However, this inhibition was not linked to tachyzoite-to-bradyzoite stage conversion. In contrast, TNF-α treatment resulted in a dramatically enhanced proliferation rate of the parasite. The cellular integrity in IFN-γ–treated organotypic slice cultures was severely impaired compared with untreated and TNF-α–treated cultures. Thus, on infection of organotypic neuronal cultures, IFN-γ and TNF-α exhibit largely detrimental effects, which could contribute to either inhibition or acceleration of parasite proliferation during cerebral toxoplasmosis.

Severe malaria is associated with the failure of host defenses to control parasite replication, with the excessive secretion of proinflammatory cytokines such as tumor necrosis factor–α (TNF-α), and with the sequestration of parasitized erythrocytes (PEs) in the microcirculation of vital organs. The scavenger receptor CD36, known as a major sequestration receptor, has also been identified as an important factor in mediating nonopsonic phagocytosis of PEs by monocytes and macrophages. The specific consequence of this phagocytosis is a decrease in parasite-induced TNF-α secretion. We evaluated the variations in CD36 level and in lipopolysaccharide (LPS)-induced TNF-α production in monocytes from Plasmodium falciparum–infected patients and in vitro in the presence of PEs. Both the monocytes from infected patients and from in vitro culture showed a decrease of CD36 expression and a reduced production of TNF-α induced by LPS. Using incubation assays with no contact between monocytes and PEs, or in the presence of a soluble supernatant obtained from the incubation of monocytes and PEs, this study shows that decreased CD36 expression was posttranscriptional and not directly related to PEs phagocytosis. In addition, these culture models suggest that the reduced capacity of TNF-α production occurred in 2 phases. The early phase (24 hr) appeared to be CD36 dependent and the second phase (48 hr) was due to a soluble factor produced by PEs. These observations suggest that the control of the TNF-α production in malaria by monocytes was not entirely dependent on the phagocytosis of PEs by CD36 and that soluble factors produced by PEs could play a role in this process.

Spauligodon zweifeli n. sp. (Nematoda: Pharyngodonidae) from the large intestines of a gecko, Lepidodactylus novaeguineae (Sauria: Gekkonidae), is described and illustrated. Prevalence of infection was 57% (mean intensity 8.5, range 1– 23). Spauligodon zweifeli n. sp. is the 43rd species to be assigned to this genus and differs from other species in the genus by possessing asymmetrical fusiform eggs in which the ends are extended as digitiform processes. It is the second species to be described from the Australian biogeographical realm.

In July 1999, 2 heteromyid rodents Heteromys desmarestianus Gray, 1868, were collected from the Área de Conservación Guanacaste, Costa Rica, and examined for parasites. Individuals of a new species of Vexillata (Travassos, 1937) Durette-Desset, 1970, were found from the intestine of hosts. Vexillata brooksi n. sp. differs from all other congeners by the presence of spicules divided at the tip instead of simple spicules. The new species closely resembles Vexillata legallae Denke, 1977, and Vexillata convoluta (Caballero and Cerecero, 1943); however, it can be distinguished from both species by possessing a smaller number of cuticular ridges at the posterior part of the body in males (9 vs. 11 and 12, respectively) and by the absence of a carene at this level.

Ascarophis valentina n. sp. is described from Mullus surmuletus off the Valencian coast of Spain on the basis of both light and scanning electron microscopy. It can be distinguished from the other members of the genus by the length of the left (long) spicule of the males and by egg morphology. An updated grouping of the species of Ascarophis considered valid is provided with respect to these characters. The new species resembles Ascarophis capelanus, belonging to the group of species possessing eggs with a single polar knob with filaments, but is distinguished by the size of the body, the length of the esophagus (especially in relation to body length), the position of the vulva, and the size of the left spicule. The new species also shows substantial morphological differences compared with the 3 species, Ascarophis mullusi, Ascarophis upenei, and Ascarophis parupenei, previously described from mullid hosts.

Species of Corynosoma (Acanthocephala) are distributed worldwide as parasites of marine mammals and sea birds. Species diagnosis is based on morphological characters, including the size and number of hooks in the proboscis and the number of spines in the dorsal and ventral regions of the body. We inferred the phylogenetic relationships of 10 nominal species of Corynosoma through analysis of internal transcribed spacers (ITS-1, ITS-2) and 5.8S ribosomal RNA sequences. Nucleotide distances between species of Corynosoma ranged from 0.4 to 11% for ITS sequences. Maximum parsimony and likelihood analyses indicated that species of Corynosoma that inhabit hosts in the marine environment form a monophyletic assemblage, but yielded conflicting hypotheses for the relationship of Corynosoma cetaceum to other members of the genus. However, parsimony and likelihood analyses were consistent for many Corynosoma sister species relationships of (e.g., C. australe plus C. bullosum, C. validum plus C. villosum, C. caspicum plus C. magdaleni, and C. enhydri plus C. strumosum). This phylogenetic framework was used to evaluate taxonomic controversies concerning C. cetaceum and C. caspicum.

Altogether, 195 fecal samples of reindeer calves (Rangifer tarandus) were collected from the ground in 3 distinct areas in eastern Iceland, where geographically isolated reindeer populations graze during the summer months. The samples were examined for coccidian parasites. Two species were found, and all infections were monospecific. Eimeria mayeri was found in calves in all sampling areas, with 1–4% prevalence and 450–167,700 oocysts per gram (opg). The sporulated oocyst lacks a polar granule but has an inconspicuous micropyle, and a small Stieda body is present on sporocysts, which are ovoid but not pointed as reported in the original description. The other coccidian, found in single calves in 2 of the 3 areas (prevalence 1 and 4%, 150 and 500 opg, respectively) is described here as a new species. The oocysts are ovoid, average 34.9 × 27.6 μm, and have 2 distinct walls. Wall thickness is ∼1.9 μm, and the outer wall, ∼¾ of total thickness, is generally smooth and appears bicolored. The outermost portion is pale red and the innermost portion yellow to pale brown. The inner wall is grey to dark brown and separated from the outer wall by a dark brown line. Oocysts contain a prominent micropyle, ∼5 μm, and enclose 4 spindle-shaped sporocysts, slightly pointed at the end opposite the Stieda body. Average size of sporocysts is 18.6 × 9.2 μm. Sporocysts contain granular sporocyst residuum and usually 2, sometimes 1, large refractile bodies in each sporozoite.

Nematodes of the subfamily Nematodirinae are characteristic components of a Holarctic fauna. The topology of a generic-level phylogenetic hypothesis, patterns of diversity, and geographic distributions for respective nematode taxa in conjunction with data for host occurrence are consistent with primary distributions determined across Beringia for species of Murielus, Rauschia, Nematodirus, and Nematodirella. Ancestral hosts are represented by Lagomorpha, with evidence for a minimum of 1 host-switching-event and subsequent radiation in the Artiodactyla. Diversification may reflect vicariance of respective faunas along with episodic or cyclical range expansion and isolation across Beringia during the late Tertiary and Quaternary. Secondarily, species of Nematodirus attained a distribution in the Neotropical region with minimal diversification of an endemic fauna represented by Nematodirus molini among tayassuids, Nematodirus lamae among camelids and Nematodirus urichi in cervids during the Pleistocene. Nematodirines are a core component of an Arctic-Boreal fauna of zooparasitic nematodes (defined by latitude and altitude) adapted to transmission in extreme environments characterized by seasonally low temperatures and varying degrees of desiccation. The history and distribution of this fauna is examined in the context of biotic and abiotic determinants for geographic colonization and host switching with an exploration of predicted responses of complex host–parasite systems to ecological perturbation under a regime of global climate change.

A relationship for Pseudostertagia bullosa within the trichostrongyloids has been enigmatic or unresolved. Studies of the synlophe in males and females of P. bullosa revealed a tapering system anterior to the deirids and a pattern of parallel ridges extending to near the caudal extremity in both lateral and median fields. Structurally, the synlophe differs considerably from that seen among the Cooperiinae and exhibits homoplasy with respect to ridge systems among some Ostertagiinae. Other structural characters due to symplesiomorphy, homoplasy or because they represent autapomorphies do not serve to reveal the putative relationships for P. bullosa with other trichostrongyloids. Although somewhat equivocal, the 2-2-1 pattern of the bursa and position of rays 2 and 3 suggest an association with the Cooperinae, as postulated by Durette-Desset and others. Pseudostertagia bullosa appears to be a species that has survived in the pronghorn, Antilocapra americana, a relictual pecoran artiodactyl that occurs in xeric regions of western North America; pronghorn are the sole remnant of the late Tertiary radiation for Antilocapridae across North America. Pseudostertagia bullosa may occur in mixed infections with a number of ostertagiines in the abomasa of mule deer (Odocoileus hemionus) and domestic sheep (Ovis aries) in regions of sympatry for pronghorn and these artiodactyl hosts.

Monophyly for the Nematodirinae, with 5 genera, Murielus, Rauschia, Nematodiroides, Nematodirus, and Nematodirella was confirmed based on comparative morphology and phylogenetic analysis of structural characters. This concept for the nematodirines excludes the monotypic Lamanema chavezi, but otherwise corroborates generic-level diversity as defined in prior studies. Exhaustive analysis resulted in 1 most parsimonious tree (36 steps; consistency index [CI] = 0.94; retention index [RI] = 0.93; excluding phylogenetically uninformative characters, CI = 0.92). As an inclusive or monophyletic group, Nematodirinae was diagnosed by 8 synapomorphies (7 are unequivocal): (1) large eggs, (2) long filiform spicules, (3) basal division of the dorsal ray, (4) symmetrical membrane enveloping the spicule tips, (5) fused structure of the spicule tips, (6) absence of the gubernaculum, (7) development of the third-stage larva within the egg, and (8) ornamentation in the form of discrete bosses on the bursa. Exclusion of Lamanema will require new assessments of historical biogeography and the evolution of host associations for the nematodirines.

Examination of the spiral intestines of 44 freshwater stingrays, Potamotrygon motoro, from tributary rivers of the Paraná River in Argentina, allowed for the collection of specimens of an undescribed species of Acanthobothrium. Acanthobothrium ramiroi n. sp. can be distinguished from all other congeners by the combination of the following characters: asymmetrical hooks (medial and lateral hooks conspicuously different in size and form, with axial prong of medial hooks stouter than abaxial prong), hook size (total length of medial hooks up to 242 μm, total length of lateral hooks up to 239 μm), bothridia not fused to the scolex proper at posterior ends, worm size (51–84 mm long), and the presence of a conspicuous vaginal sphincter. The new species is different from all other species of Acanthobothrium in freshwater potamotrygonids, except Acanthobothrium terezae, in having conspicuous asymmetrical hooks. The main differences that allow for the distinction between A. ramiroi and A. terezae include hook size, the way the bothridia are attached to the scolex proper, and the shape of the older gravid segments. The discovery of a new species of Acanthobothrium from a potamotrygonid extends our understanding of the diversity of the genus in freshwater stingrays in South America.

Five species of Leucocytozoon were recovered from 35/828 birds of 95 species examined from 6 sites in West Africa between May 1995 and June 2001. Leucocytozoon pogoniuli n. sp. is described from the tinker barbets Pogoniulus subsulphureus and Pogoniulus atroflavus. Leucocytozoon trachyphoni n. sp. is described from the barbet Trachyphonus purpureus. No leucocytozoids have been reported previously in species of Pogoniulus. Leucocytozoon nectariniae was identified from the sunbird Nectarinia olivacea, and Leucocytozoon brimonti was recovered from 4 species of Pycnonotidae (bulbuls), all of which are new host records. We also report the first Leucocytozoon to be recovered from the phylogenetically isolated bird, Picathartes sp. (Picathartidae). This parasite is similar in appearance to Leucocytozoon sakharoffi, and probably represents a previously undescribed species. In view of the intraspecific variability and, frequently, relatively minor interspecific differences within Leucocytozoidae, we suggest that the development and application of molecular techniques would greatly advance understanding of speciation and relationships within this family.

Gorgoderina parvicava, G. diaster, and G. megacetabularis n. sp. are reported inhabiting the urinary bladders of Rana vaillanti and R. cf. forreri from northwestern Costa Rica. Gorgoderina megacetabularis n. sp. differs from all other species of the genus by the combination of the following characters: small body size (2.78–3.17, mean 2.92 mm), sucker ratio (1:3.1– 3.7), and by the presence of 2 compact, oval, unlobed vitelline masses. Redescription of G. diaster including previously undescribed details on the reproductive apparatus and morphometric data is provided. This is the first record of the 3 species of Gorgoderina in Costa Rica and is the first record of G. diaster in R. vaillanti and R. cf. forreri.

A new species of trichosomoidid nematode, Huffmanela canadensis n. sp., is described from the skin of rockfish (Sebastes spp.) from the Pacific Ocean off the coast of Vancouver Island (Clayoquot Sound region), British Columbia, Canada, on the basis of the morphology of the adult worms and their eggs in the host's tissue and the biological characters. The species is characterized mainly by the shape and structure of the fully developed eggs (absence of surface envelope, surface with transverse ridges), by their small size (48–63 × 24–27 μm), and by the site of infection (skin). Besides Huffmanela huffmani, this is the second Huffmanela species in which adult worms are known in addition to the eggs from the host's tissues (most Huffmanela spp. have been described only from their conspicuous eggs occurring in various tissues of fishes). Adults of H. canadensis differ morphologically from those of H. huffmani, mainly in the structure of the male caudal end and in the distinctly elevated anterior vulvar lip of the female.

Helminthological examination of specimens of the freshwater fishes Bryconamericus iheringi Boulenger, 1887 (Characidae) and Jenynsia multidentata (Jenyns, 1842) (Anablepidae), collected from Medina River, Province of Tucumán, Argentina, revealed the presence of 2 species of parasitic nematodes, Rhabdochona fabianae n. sp. and Rhabdochona acuminata (Molin, 1860) (Rhabdochonidae). Rhabdochona fabianae n. sp. was characterized by the presence of 16 anterior teeth in the prostom and filamented eggs in females. Rhabdochona acuminata was recorded for the first time in the north of Argentina.

DNA was extracted and the 18S ribosomal RNA gene was amplified and sequenced from the blood of 2 Pallas' cats (Otocolobus manul) infected with small intraerythrocytic piroplasms. Sequences of the parasite were found to be identical with that of a previously reported Cytauxzoon-like piroplasm from a Pallas' cat. Phylogenetic analyses of the parasite DNA sequences obtained from the 3 Pallas' cats to other piroplasms revealed a sister group relationship to C. felis. The mean corrected percent sequence divergence between the Pallas' cat parasite and C. felis was 1.490%, which is greater than that for most other piroplasms in which species status has been accepted. On the basis of the sequence variation, we propose to name the Pallas' cat parasite C. manul. Phylogenetic analyses of C. manul also revealed a close relationship with the Spanish Cytauxzoon-like isolate because they exhibited only 0.389% sequence divergence, yet these sequences exhibit a mean of 1.690% sequence divergence from the New World isolate of C. felis. Our phylogenetic analyses also revealed several taxonomic problems that have impeded the development of a classification that accurately reflects evolutionary history of piroplasms. As currently arranged, Babesia and Theileria are paraphyletic taxa and are in need of reorganization.

Dissonus hoi n. sp. is described from female and male specimens collected from the nasal cavities of a Samson Fish (Seriola hippos Günther) captured off Rottnest Island, Western Australia. Adult female D. hoi are distinguished from their congeners by possessing the following combination of characters: (1) a quadrangular genital complex, (2) ventral spines on the genital complex, (3) pair of postantennal processes, (4) a sternal furca, (5) 1-segmented abdomen, and (6) convoluted, uniseriate egg strings. New records and intraspecific morphological variation are also given for Dissonus nudiventris collected from Australia, and Dissonus similis collected from the tropical western Pacific. A key to the valid species of Dissonus is provided.

A new species of Parahadjelia occurs in the muscovy duck, Cairina moschata (Linnaeus, 1758), from the Area de Conservacion Guanacaste, Costa Rica. The new species differs from Parhadjelia neglecta Lent and Freitas, 1939, in the body size, in the spicules shape and ratio of spicules, and in having 2 pairs of sessile papillae near the tail tip of the male. The characters exhibited by P. neglecta and the new species validate its generic status distinct from Hadjelia.

Two species of Skrjabinura Gnedina, 1933, were collected in the intestines of birds from the Area de Conservación Guanacaste (ACG), Costa Rica. Skrjabinura mesoamericana n. sp. inhabits Dendrocincla homochroa, Calocitta formosa, Dendrocolaptes certhia, Basileuterus rufifrons, and Chordeiles acutipennis. The new species differs from all species of the genus by having dissimilar spicules, the right having a distinctive thin and bent handle on the proximal end. The new species can be further distinguished from Skrjabinura pomatostomi and Skrjabinura brevicaudatum by having subequal versus equal spicules. Skrjabinura mesoamericana resembles S. brevicaudatum in the number and arrangement of male caudal papillae but differs in the vulva position and in having smaller eggs. The new species differs from S. potamostomi in having 6 versus 8 pairs of postcloacal papillae. Among those species having subequal spicules, the new species further differs from Skrjabinura spiralis in having 1 pair of paracloacal papillae and 6 pairs of postcloacal papillae, versus no paracloacal papillae and 4 pairs of postcloacal papillae, and in the vulva position. The new species is perhaps most similar to Skrjabinura vali, from which it further differs by having 3 pairs of precloacal, 1 pair of paracloacal, and 6 pairs of postcloacal versus 2 pairs of precloacal, 2 pairs of paracloacal, and 6 pairs of postcloacal papillae in the vulva position and in the shape of the eggs. Skrjabinura vali (Guerrero, 1971) Chabaud, 1978, originally described in Piaya cayana from Venezuela, occurs in the small intestine of P. cayana, as well as the new hosts, Crotophaga sulcirostris and Myiarchus tyrannulus, in the ACG, a new locality. Our specimens differ from the original description in the body length of the female, the numbers of postanal papillae of male tail, and the size of eggs.

Neospora hughesi is a recently identified cause of equine protozoal myeloencephalitis. However, the significance of this parasite is poorly understood. An enzyme-linked immunosorbent assay (ELISA) with a recombinant form of the N. hughesi 29-kDa surface antigen (rNhSAG1) was developed for serodiagnosis of equine N. hughesi infections. Parallel ELISA analysis showed that animals immunized or infected with N. hughesi exhibited greater antibody reactivity with rNhSAG1 than with the Neospora caninum homolog, rNcSAG1. The rNhSAG1 ELISA showed 94.4% sensitivity and 95.0% specificity when compared with N. hughesi western blot results for 1,006 samples. The N. hughesi seroprevalence was 3.4% for the 1,917 samples tested by ELISA, which is less than earlier reports. Importantly, western blot analysis of ELISA-positive sera revealed only 18 true seropositive samples for an even lower seroprevalence of 0.9%. These results imply that Neospora spp. infections are uncommon in horses. The sensitivity and specificity exhibited by the rNhSAG1 ELISA suggest that it has a potential use for serodiagnosis of N. hughesi infection in equids. Furthermore, the high-throughput capability of the ELISA will allow for screening large sample sets, which should provide a better understanding of N. hughesi epidemiology.

The ookinete is the key determinant of infection within the mosquito vector, yet there are few population studies of ookinetes in nature. This investigation compared different techniques used to estimate ookinete densities in mosquitoes. Laboratory-reared Anopheles dirus mosquitoes were fed on gametocytemic blood drawn from 7 Plasmodium vivax patients at a malaria clinic in Mae Sot, Thailand. At 20–26 hr, bloodmeals were excised. Three techniques were evaluated, i.e., hemacytometer counts under phase-contrast microscope, Giemsa staining of bloodmeal smears, and immunofluorescent staining with a monoclonal antibody specific against the 25-kDa antigen expressed on the surface of P. vivax zygotes and ookinetes. Additional mosquitoes were dissected at day 10 for oocysts. The hemacytometer method was the simplest and quickest method but lacked precision at low ookinete densities. Immunofluorescent staining was the most sensitive, accurate, and the only method that enabled unequivocal detection of zygotes. Bloodmeals contained a mixture of zygotes, retorts, and mature ookinetes, indicating that postzygotic development of P. vivax in A. dirus was asynchronous. The conversion efficiency of zygotes/ookinetes to oocysts varied among patients and was independent of zygote-ookinete density, suggesting that variations in host blood composition, e.g., antibodies, drugs, etc., may influence the success of zygote-ookinete development.

Cytauxzoon felis produces a disease in domestic cats in the Midwest (U.S.A.), which often leads to a fatal outcome. Although the clinical disease process is well described, there are still many unanswered questions about this organism. For example, it is unknown whether species of ticks other than Dermacentor variabilis can serve as vectors for transmission. With recent reports of surviving cats from limited geographic areas, another relevant question is the potential for genetically less virulent organism strains. This study evaluated 352 individual or pooled tick samples (1,362 total ticks) for the presence of C. felis small subunit ribosomal RNA and internal transcribed spacer 1 (ITS-1) region genes using a polymerase chain reaction (PCR). These ticks were collected from dogs and cats in several Missouri counties, including 10 from cats diagnosed with cytauxzoonosis. Only 3 positive C. felis samples were identified in Amblyomma americanum nymphs, and there was very limited genetic variation noted in both genes. The small number of positive samples did not allow the study to determine which PCR analysis was more sensitive. This is the first known report of ITS-1 gene identification and sequencing for C. felis. It is also the first published investigation of genetic variation in C. felis.

Thirty-three splenectomized Aotus lemurinus griseimembra monkeys with no previous experience with malaria were infected with the Vietnam Palo Alto strain of Plasmodium vivax. The median maximum parasite count was 280,000/μl. Nine splenectomized monkeys with previous infection with Plasmodium falciparum had median maximum parasite counts of 120,000/μl. Splenectomized Aotus nancymai monkeys supported infections at a lower level. Transmission via the bites of Anopheles dirus mosquitoes was obtained in a splenectomized A. lemurinus griseimembra, with a prepatent period of 31 days. It is estimated that between 1.5 × 10⁸ and 1.6 × 10⁹ parasites can be removed from an infected animal for molecular or diagnostic antigenic studies.

New cases of human babesiosis were recently reported in Europe. The etiological agent was identified as Babesia EU1, a zoonotic pathogen with previously unreported molecular characteristics. On the basis of a comparison of the complete babesial 18S rRNA gene, we have generated strong molecular evidence that Ixodes ricinus ticks from Slovenia are infected with EU1.

We analyzed the prevalence, intensity, and medium density of parasitism of Hesperoctenes fumarius infesting Molossus rufus in natural (hollow trees) and anthropogenic roosts (attics) in southeastern Brazil. The prevalence and intensity of infestations were higher in the hollow trees than in the attic roosts. We also noted a relationship between the amount of space available within the roost and the infestation levels of H. fumarius. One advantage of roosting in larger, often man-made, refuges may be the reduction in ectoparasite infestations.

Mechanisms of host immunosuppression after infection with Toxoplasma gondii are unclear. This study was performed to observe cytokine and immunoglobulin secretions by murine splenic lymphocytes infected in vitro with live, nonreplicating (irradiated) RH tachyzoites on stimulation with concanavalin A (Con A) or lipopolysaccharide (LPS). For lymphocyte cultivation, 3 groups were prepared: coculture with live nonirradiated tachyzoites separated by a transwell (group T), live irradiated tachyzoites without a transwell (group R), and no tachyzoites (group C). Compared with group T, groups R and C, on stimulation with Con A, revealed significantly (P < 0.05) lower levels of interleukin-2 (IL-2) and IFN-γ, but not IL-10. The levels of IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM were also significantly (P < 0.05) lower in groups R and C than in group T after stimulation with LPS. The results suggest that intracellular infection of murine splenic lymphocytes with T. gondii tachyzoites could impair their capacity to produce cytokine and immunoglobulin secretions.

Raccoons (Procyon lotor) are reservoir hosts for Trypanosoma cruzi. A 3-yr-long serological survey was conducted to determine the prevalence of antibodies to this zoonotic parasite in raccoons collected from Fairfax County, Virginia, a suburban/urban area outside Washington, D.C. Serum samples from 464 raccoons were examined for T. cruzi antibodies at a 1:40 dilution with an indirect fluorescent antibody test (IFAT) and Brazil strain T. cruzi amastigotes and trypomastigotes as antigen. A positive IFAT test was found in 154 (33%) of the 464 samples. The yearly prevalence was 49 of 132 (37%) in 2000; 19 of 120 (16%) in 2001; and 86 of 212 (41%) in 2002. Our study indicates that raccoons in this area of Virginia are frequently exposed to T. cruzi.

The susceptibility of wild-caught European passeriform birds to naturally isolated malaria parasites, Plasmodium (Novyella) nucleophilum and Plasmodium (Novyella) vaughani, was studied by means of intramuscular subinoculation of infected citrated blood. Plasmodium nucleophilum of the great tit, Parus major, was transmitted to 3 great tits, but 3 blackcaps (Sylvia atricapilla) were not susceptible. Plasmodium vaughani of the robin, Erithacus rubecula, was transmitted to 1 robin and 1 blackcap, but 1 dunnock, Prunella modularis, was not susceptible. The prepatent period was between 8 and 10 days in all experimental infections. Maximum experimental parasitemia (3.4% of red cells) was detected in great tits infected with P. nucleophilum 23 days postexposure. A light (<0.01%) transient parasitemia of P. vaughani developed in the robin and blackcap. This study is in accord with former experimental observations on host specificity of P. nucleophilum and P. vaughani, which are characterized by a wide, but selective, range of avian hosts. Two new host–parasite associations were recorded.

Of the 18 trematode species that use the horn snail, Cerithidea californica, as a first intermediate host, 6 have the potential to use raccoons as a final host. The presence of raccoon latrines in Carpinteria Salt Marsh, California, allowed us to investigate associations between raccoons and trematodes in snails. Two trematode species, Probolocoryphe uca and Stictodora hancocki, occurred at higher prevalences in snails near raccoon latrines than in snails away from latrines, suggesting that raccoons may serve as final hosts for these species. Fecal remains indicated that raccoons fed on shore crabs, the second intermediate host for P. uca, and fish, the second intermediate host for S. hancocki. The increase in raccoon populations in the suburban areas surrounding west coast salt marshes could increase their importance as final hosts for trematodes in this system.

In this study, P23 of Cryptosporidium parvum sporozoites, an immunodominant surface protein, was stably expressed in Toxoplasma gondii (Tg/P23) and its protective effects were evaluated in a mouse model. The molecular weight and antigenic property of P23 expressed by Tg/P23 were similar to those of the native P23. Mice immunized with lysed Tg/P23 tachyzoites produced specific neutralizing antibodies against C. parvum. These findings indicate that the T. gondii vector may provide a new tool for the production of a recombinant vaccine against cryptosporidiosis in animals.

Abundant, apparently normally developing, liver-stage parasites of Plasmodium coatneyi were demonstrated following injection of sporozoites dissected from the salivary glands of Anopheles dirus mosquitoes. Erythrocytic development was not demonstrated.