The effect of the trematode Microphallus turgidus on its second intermediate host, the grass shrimp, Palaemonetes pugio, was tested. To do so, we measured the susceptibility of infected and uninfected shrimp to predation by the mummichog, Fundulus heteroclitus. Shrimp behavior was compared in the presence and absence of a fish predator, and the swimming stamina and backthrust escape responses of infected and uninfected shrimp were measured. Infected shrimp were more likely to be eaten by a predator than uninfected shrimp, had lower swimming stamina, and spent more time swimming and less time motionless in the presence of a predator. There was no difference between backthrust distances traveled in response to a stimulus by either infected or uninfected shrimp. Thus, M. turgidus may increase the predation of P. pugio in the wild, possibly by affecting the swimming stamina and predator avoidance responses of the shrimp.

Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate–conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov–Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose–mannose and N-acetyl-d-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-d-galactosamine–specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (l-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent.

Cytotoxic-activated macrophages control Toxoplasma gondii growth by producing nitric oxide (NO). However, the parasite can partially inhibit NO production. NO is generated from arginine within the polyamine biosynthetic pathway. Two enzymes of this pathway are ornithine decarboxylase (ODC) and arginine decarboxylase (ADC). The aim of the present work was to investigate whether T. gondii is able to modulate polyamine metabolism in macrophages. Toxoplasma gondii infection did not affect basal ODC or ADC activity. However, lipopolysaccharide induced an increase in ODC activity. Polyamine-treated macrophages exhibited a T. gondii–infection index similar to controls but a higher adhesion index; the parasite did not grow in methyl-ornithine (ODC inhibitor)–treated macrophages. The parasites were able to take up putrescine with a K_m of 0.92 μM, indicating the presence of a high-affinity putrescine-transporter system. Putrescine-treated T. gondii actively penetrated macrophages and Vero cells. However, NO production and lysosomal parasitophorous vacuole fusion were not inhibited. Considered together, these results demonstrate that T. gondii requires polyamines for multiplication. However, as opposed to Trypanosoma cruzi and because of a relatively high-affinity putrescine-transporter system in the parasite, constitutive macrophage levels of putrescine seem sufficient to support T. gondii survival and multiplication.

Equine protozoal myeloencephalitis is a major cause of neurological disease in horses from the Americas. Horses are considered accidental intermediate hosts. The structure of sporocysts of the causative agent, Sarcocystis neurona, has never been described. Sporocysts of S. neurona were obtained from the intestines of a laboratory-raised opossum fed skeletal muscles from a raccoon that had been fed sporocysts. Sporocysts were 11.3 by 8.2 μm and contained 4 sporozoites. The appearance of the sporocyst residuum was variable. The residuum of some sporocysts was composed of many dispersed granules, whereas some had granules mixed with larger globules. Excystation was by collapse of the sporocyst along plates. The sporocysts wall was composed of 3 layers: a thin electron-dense outer layer, a thin electron-lucent middle layer, and a thick electron-dense inner layer. The sporocyst wall was thickened at the junctions of the plates. Sporozoites were weakly motile and contained a centrally or posteriorly located nucleus. No retractile or crystalloid body was present, but lipidlike globules about 1 μm in diameter were usually present in the conoidal end of sporozoites. Sporozoites contained 2–4 electron-dense rhoptries and other organelles typical of coccidian zoites. Sporozoites entered host cells in culture and underwent schizogony within 3 days.

Neospora caninum is an apicomplexan parasite identified as a major cause of abortion in cattle and neurological disease in various animal species. It is closely related to Toxoplasma gondii, sharing the ability to persist indefinitely in latent stage within the host as a tissue cyst containing slow-dividing bradyzoites. In this study, we compared different stress methods to induce in vitro bradyzoite conversion, using MARC-145 cells infected with Nc-Liverpool isolate. The tachyzoite-to-bradyzoite conversion rate was monitored at days 3, 5, and 7 after stress in a double-immunofluorescence assay using a monoclonal antibody against the tachyzoite antigen SAG1 (αSAG1) and a rabbit serum directed to the intracytoplasmic bradyzoite antigen BAG1 (αBAG1). Seven days of treatment with 70 μM sodium nitroprusside offered the highest bradyzoite transformation rate and the best yield of total parasitophorous vacuoles observed. In the present work, we introduce an alternative, simplified, and more advantageous method for bradyzoite production of N. caninum, using a reliable cell culture system easy to handle and with promising capacity of parasite purification.

This article examined the composition of parasite assemblages of whitefish (Coregonus lavaretus) in 8 interconnected lakes in northeastern Finland and evaluated the role of coregonid specific parasites and the geographical distance between populations in determining the similarity of the assemblages. Parasite assemblages were compared using the Jaccard qualitative similarity index and a quantitative similarity index and by incorporating the allogenic–autogenic species concept and the effects of 2 corresponding measures of geographical distance between the lakes. The majority of the parasite species found (10 of 14) were specific to salmonids. Similarity of assemblages of autogenic parasites between the lakes was negatively correlated with geographical distance. The dominance of 2 parasite species, the whitefish specialist Ichthyocotylurus erraticus and the generalist Ergasilus sieboldi, was also demonstrated. We concluded that the high proportion of widespread parasite species specific to coregonids is an important determinant of similarity in these assemblages. However, ecological factors were likely to contribute to qualitative (presence of species) and quantitative (difference in abundances) differences between lakes, in the case of autogenic parasites, their importance increasing with geographical separation.

The diagnosis of Kudoa funduli (Hahn, 1915) Meglitsch, 1948 (Myxozoa), is supplemented through study of new material collected from Fundulus heteroclitus (Cyprinodontidae) in coastal waters of Nova Scotia, Canada, and Connecticut. Plasmodia normally develop intracellularly in striated muscle of the flank and head, eventually rupturing and releasing spores. Spores disperse along adjacent epimysium, sometimes as far as the skin surface. Some plasmodia develop extracellularly within the bony cavities of vertebrae. Formalin-fixed spores viewed with a light microscope possess rounded edges, an inconspicuous apical region, thin sutural ridges, measure 6.6–7.4 μm wide, 4.3–5 μm thick, and 5.1–5.4 μm long, and have 4 equally sized polar capsules, 1.7–2.3 μm length by 1.4–1.7 μm width. Scanning electron microscopy revealed that spores are almost stellate, with inconspicuous uplifted tips, and that, within intracellular plasmodia, are embedded in an extensive honeycomb-like matrix. Prevalence of infection of K. funduli was 100% in host populations sampled in both Nova Scotia and Connecticut. Molecular sequence data of the 18S ribosomal DNA (737 base pairs) reveal that K. funduli is a valid species and a member of a clade that includes Kudoa dianae Dyková, Avila, and Fiala, 2002, Kudoa miniauriculata Whitaker, Kent, and Sakanari, 1996, and Kudoa paniformis Kabata and Whitaker, 1981.

Schistobrachia jordaanae n. sp. (Copepoda: Siphonostomatoida: Lernaeopodidae) is described from adult female specimens collected from gill filaments of a diamond ray Gymnura natalensis (Gilchrist and Thompson, 1911) captured in the Indian Ocean off the South African coast. Schistobrachia jordaanae is best distinguished from its congeners by 2 unique characteristics: it possesses a necklike region between the origin of its maxillae and maxillipeds and the tips of its maxillae bifurcate repeatedly to form a rootlike anchor.

Gray squirrels, Sciurus carolinensis, were livetrapped in 2 different habitat types, woodland (67 squirrels) and parkland (53 squirrels), in southeastern Georgia. Ectoparasites were recovered from anesthetized squirrels and compared between hosts from the 2 habitats. Because of the absence of low vegetation in parkland habitats, it was hypothesized that the ectoparasite fauna, especially ticks and chiggers, would be more diverse on woodland squirrels. The results were generally in agreement with this hypothesis. Seventeen species of ectoparasites were recovered from woodland squirrels, compared with 6 species from parkland squirrels. Five species of ticks and 3 species of chiggers parasitized the woodland squirrels compared with no ticks or chiggers on the parkland squirrels. Significantly higher infestation prevalences were recorded on woodland compared with parkland squirrels for the flea Orchopeas howardi, the tick Amblyomma americanum, and the mesostigmatid mite Androlaelaps fahrenholzi. The mean intensity for O. howardi also was significantly higher on woodland than on parkland squirrels. Because a new strain of Bartonella sp. was isolated recently from S. carolinensis in Georgia, selected ectoparasites from this study were screened for bartonellae by polymerase chain reaction (PCR). Some of the fleas and lice, but none of the mites tested, were PCR positive, suggesting that fleas, or lice, or both, might be vectors of bartonellae between squirrels. Six distinct strains of Bartonella sp. were detected, 2 in fleas and 4 in lice.

Three new species of Antricola (Acari: Argasidae) are described from adult specimens collected on bat guano in different caves in Brazil. The female of Antricola guglielmonei n. sp. is easily determined by the presence of 2 smooth, depressed areas lacking setae in the posterolateral portions of the dorsum of the idiosoma, together with the partial fusion of the tubercles in the anteromedian portion of the idiosoma. The male of this species has a small spiracular plate surrounded by a pattern of tubercles disposed concentrically in its dorsal portion. Both sexes have cervical grooves very slightly marked. Antricola delacruzi n. sp. represents the only Antricola species with the dorsum of the idiosoma devoid of tubercles in both sexes and with scarce and minute setae placed over the smooth cuticle. The female of A. inexpectata n. sp. is known only from a few specimens. In this species, lines of smooth cuticle lacking setae separate the tubercles of the dorsum. In addition, there are 3 clumps of plumose setae close to the spiracular plate, in the ventrolateral portions of the idiosoma, in 3 well-delimited regions over cuticular thickenings. These 3 species share the peculiarity of a Haller's organ with the anterior pit bearing only 7 2 setae. The collection of these new species in Brazilian caves greatly expands the known range of the genus. A key to the adults of all known species of the genus is provided.

Here, for the first time in an ascaridoid (Hysterothylacium auctum), we present structural features of the phasmids, paired sense organs, positioned in a bilateral manner close to the point of the tail; the features were obtained using scanning and transmission electron microscopy. We found that each phasmid consists of a single ciliated dendritic process situated in a phasmidial canal surrounded by 2 supporting cells, a socket and a sheath cell. The socket cell contains clusters of electron-dense fibrous material in its apical region and covers the phasmidial canal along its whole length. The sheath cell is characterized by a well-developed endoplasmic reticulum. The phasmidial canal is lined with a thin layer of cuticle that becomes incomplete at the base of the ciliated dendritic process. In this region, the dendritic process consists primarily of a high number of microtubule singlets and some peripheral microtubule doublets. The base of the dendritic process, containing numerous striated rootlets, gives off a large number of fingerlike offshoots, villi, invading the surrounding sheath cell. The systematic significance and functional implication of the phasmid in nematodes are also discussed.

Mating system plays a determinant role in the maintenance and distribution of genetic variations. It can be assessed indirectly by analyzing the distribution of the genetic variability within populations or directly by considering how mating pairs are formed. In the present study, 71 pairs of adult Schistosoma mansoni worms sampled from naturally infected rats were genotyped to investigate how male and female schistosomes paired according to their genetic relatedness. Among all samples, pangamy, the random association between males and females, could not be rejected. Whereas the schistosome mating system has been intensively studied under experimental conditions, to the best of our knowledge, our study is the first to attempt to understand the way in which males and females pair in natural conditions.

In the present study, we compared the effect of 5% oxygen tension (hypoxia) with a normal tension of 21% oxygen (normoxia) on macrophage infection by the protozoan parasite Leishmania amazonensis. Macrophages from different sources (human cell line U937, murine cell line J774, and murine peritoneal macrophages) exposed to hypoxia showed a reduction of the percentage of infected cells and the number of intracellular parasites per cell. Observations on the kinetics of infection indicated that hypoxia did not depress L. amazonensis phagocytosis but induced macrophages to reduce intracellular parasitism. Furthermore, hypoxia did not act synergistically with γ-interferon and bacterial lipopolysaccharides in macrophages to induce killing of parasites. Experiments also indicated no correlation between nitric oxide production and control of infection in macrophages under hypoxic condition. Thus, we have provided the first evidence that hypoxia, which occurs in various pathological conditions, can alter macrophage susceptibility to a parasitic infection.

Challenge of 1-yr Trypanosoma cruzi chronically infected mice with trypomastigotes results in a consistent reduction of parasite dissemination that correlates with spleen activation and increase in the anti–T. cruzi effector immune mechanisms. That is, parasite challenge results not only in elimination of the inoculum but also in a drastic decrease in basal subpatent parasitemia levels as revealed by transferring blood samples to immunosuppressed mice. Parasite elimination correlated with (1) a brief and intense burst in the ability of spleen cells to produce interferon-γ, (2) an increase in total IgG2a-producing spleen cells, (3) higher parasite-specific IgG2a serum levels, and (4) an accumulation of non–B, non–T class II cells in the spleen. Furthermore, challenged, chronically infected mice had increased numbers of B, CD4 , and CD8 large spleen cells. Besides reinforcing the activation of protective Th1 effector mechanisms, challenge with T. cruzi also induced Th2 effector molecules, such as interleukin (IL)-10 and IL-4, and IL-4–dependent IgG1. Our results are the first evidence that the immune system of T. cruzi chronically infected mice can be optimized in its ability to restrict parasite dissemination, opening the possibility that therapeutic vaccination could be used to reduce the parasite load and pathology of patients with chronic Chagas' disease.

Trypanosoma cruzi, a protozoan parasite, is the etiologic agent of Chagas disease. The disease is characterized by acute and chronic phases, with high and low parasitemia, respectively. A strong immune activation is necessary for the host to enter the chronic phase; however, immune mechanisms participating in the reduction of parasites between the acute and chronic phases of the disease have been very difficult to elucidate. We report here the discovery of anti-egressin, an antibody present in serum from chronically infected BALB/c mice that is able to inhibit parasite egress from infected BALB/c fibroblast cultures in vitro. The antibody is very concentrated in serum from these mice; chronic serum may be diluted 1:20 while still maintaining functional activity. Isotype analysis of anti-egressin has suggested it to be IgG2a. Further analysis revealed that anti-egressin was specific for a component expressed on the surface of infected host cells. The specificity of anti-egressin toward the extracellular portion of infected host cells was demonstrated both by using a quantitative assay measuring released trypomastigotes and through immunocytochemical staining. The novel role of anti-egressin in the inhibition of parasite egress from infected host cells has not been described in the literature to date. We believe that anti-egressin plays an important role in achieving the low parasitemia characteristic of chronic Chagas disease.

A review of current literature on mammalian hosts' sexual dimorphism (SD) in parasitic infections revealed that (1) it is a scarcely and superficially studied biological phenomenon of considerable significance for individual health, behavior, and lifestyles and for the evolution of species; (2) there are many notable exceptions to the rule of a favorable female bias in susceptibility to infection; (3) a complex network of molecular and cellular reactions connecting the host's immuno-neuro-endocrine systems with those of the parasite is responsible for the host–parasite relationship rather than just an adaptive immune response and sex hormones; (4) a lack of gender-specific immune profiles in response to different infections; (5) the direct effects of the host hormones on parasite physiology may significantly contribute to SD in parasitism; and (6) the need to enrich the reductionist approach to complex biological issues, like SD, with more penetrating approaches to the study of cause–effect relationships, i.e., network theory. The review concludes by advising against generalization regarding SD and parasitism and by pointing to some of the most promising lines of research.

Periovular granulomas are the major lesions in baboons infected with Schistosoma mansoni. Temporal Northern blot analysis of cytokine messenger RNA (mRNA) expression in granulomatous baboon livers demonstrated tissue-specific expression. Interleukin 1 beta (IL-1β), IL-6, tumor necrosis factor alpha, and migration inhibitory factor (MIF) mRNAs were expressed strongly at week 6 of infection and decayed thereafter, whereas interferon gamma (IFN-γ), IL-2, IL-10, and IL-12 mRNAs were first expressed at week 12, with IFN-γ and IL-12 mRNA expression persisting until week 17. IL-4 and IL-5 mRNAs first appeared at week 12, with IL-4 persisting unchanged and IL-5 increasing by week 17. Thus, egg deposition induced strong hepatic expression of proinflammatory and downregulatory cytokines. The cooccurrence of IL-2, IFN-γ, IL-4, and IL-5 mRNAs at week 12 confirms that baboons, like humans, show a mixed type 1–type 2 cytokine response. When granulomas had become smaller at 17 wk, IFN-γ, IL-4, and IL-5 were the only cytokine mRNAs that were expressed strongly, implicating them in granuloma modulation. The early expression of MIF mRNA and MIF's role as the main counterregulator of glucocorticoid immunosuppression ties in with our earlier demonstrations of circulating adrenal steroids changing with the progression of schistosomiasis in baboons and of proinflammatory cytokine mRNA expression in the hypothalamic–pituitary–adrenal axis tissues of infected baboons. Together, these data imply neuroendocrinological influences on disease progression in schistosomiasis.

Leishmaniasis is caused by the protozoan parasite Leishmania spp. In murine leishmaniasis, a T helper cell type-I (Th1) response, characterized by the secretion of interferon (IFN)-γ is necessary for clearing the infection, whereas a Th2 response, accompanied by the production of interleukin (IL)-5, can exacerbate the disease. Moreover, the early cytokine milieu is thought to play an important role in determining the outcome of infection. In human leishmaniasis little is known about this early cytokine response. Because of this, we cocultured human peripheral blood mononuclear cells (PBMC) with Leishmania major in vitro and measured the production of IFN-γ, IL-5, and IL-10. We also treated PBMC cultures with various cytokines and neutralizing anticytokines. We found that the principal cytokine produced was IFN-γ and that its production was regulated by IL-10 and IL-12. In contrast, only low levels of Th2 cytokines such as IL-5 were produced. Therefore, the Th1–Th2 dichotomy that exists in inbred strains of mice does not appear to apply to the response of humans to L. major. Rather, Th2 cytokines may play a role in regulating IFN-γ production.

The parasite faunas of juvenile English sole (Parophrys vetulus) in 1971–1972 and staghorn sculpin (Leptocottus armatus) in 1971 from Yaquina Bay, Oregon, were compared with faunas found in the same estuary in 1997–2000 (English sole) and 1999–2000 (staghorn sculpin). The 7 most commonly occurring parasites in 1971 were compared with the same species observed during the same month and sampling sites in 1997–2000. Multivariate community analysis of juvenile English sole parasites supported the suggestion that the 1971 parasite data were representative of the early-1970s time period. Four of the parasite species infecting English sole and 6 of those infecting staghorn sculpins had significantly lower prevalences in 1997– 2000. Parasite species with significantly lower prevalences also had reduced intensity levels. One parasite (Glugea stephani) of English sole increased in prevalence in the 1997–2000 samples in association with the warm estuarine temperatures during the 1997 El Niño year. Although the causes for the changes in occurrence of other parasites were not determined, ecological changes in Yaquina Bay that may have influenced parasite ecology include apparent changes in the estuary ichthyofauna that occurred between the sampling periods. Such changes could be associated with increases in the number of California sea lions (Zalophus californianus) subsequent to establishment of the Marine Mammal Protection Act in 1972.

The helminths of 218 white-toothed shrews from 29 sites in 2 biotopes in the Albufera Natural Park (Valencia, Spain) were examined from July 1990 to August 1991. An association analysis of helminths occurring at a prevalence of more than 4% was carried out for 4 species of cestodes located in the intestine (Hymenolepis pistillum, H. scalaris, H. tiara, and Pseudhymenolepis redonica) and 3 species of nematodes (Pseudophysaloptera sp. located in the stomach, Stammerinema rhopocephala larvae in the intestine and abdominal cavity, and Porrocaecum sp. in the thoracic and abdominal cavities). Bivariate (species pairs) versus multivariate analyses (associations within the entire set of species) were performed of presence–absence and of quantitative records (influence of intensity on associations). The associations were evaluated with respect to the sex and age of the host and to the sampling date and sites. The host and environment played a limited role, and the major determinant of species assemblage was phylogenetic. Positive associations were found among both the cestodes and the nematodes, whereas negative associations were found between cestodes and nematodes. The type of life cycle was probably the second greatest determinant of species associations. Nematodes using shrews as a paratenic host or as their definitive host were both positively associated.

Here, we analyzed histological findings and parasite burden in chronic Neospora caninum infection in BALB/c and ICR mice and studied the correlation between lesion severity and parasite load in brain. To obtain a better understanding of the infection, we examined the influence of various host–pathogen factors. Groups of outbred (ICR) and inbred (BALB/c) mice were inoculated using several NC-1 parasite doses (4 × 10⁵, 10⁶, and 5 × 10⁶ tachyzoites), inoculation routes (intraperitoneal and subcutaneous), and 3 immunosuppressive treatments (methylprednisolone, cyclophosphamide, and vinblastine). Lesion severity was analyzed in the liver, lung, heart, and brain tissues, and parasite load was measured by real-time polymerase chain reaction in brain tissue. The results indicated more severe cerebral lesions and higher brain parasite burdens in inbred than in outbred mice. Hepatic tissue was the primary lesion site in immunosuppressed ICR mice. We also observed that increased inoculum size was reflected in greater lesion severity and a higher cerebral parasite load. No difference was observed with respect to inoculation route. The study also showed an association between brain parasite burden and severity of cerebral lesions in BALB/c mice.

Kudoa hypoepicardialis n. sp. infects the space between the epicardium and the compact myocardium and, in intense infections, the pericardial chamber of man-of-war fish (Nomeus gronovii) (Nomeidae) (the type host), blue runner (Caranx crysos) (Carangidae), Warsaw grouper (Epinephelus nigritus) (Serranidae), Atlantic tripletail (Lobotes surinamensis) (Lobotidae), northern red snapper (Lutjanus campechanus) (Lutjanidae), black drum (Pogonias cromis) (Sciaenidae), and bluefish (Pomatomus saltatrix) (Pomatomidae) in the northern Gulf of Mexico. This is the first report of a Kudoa sp. from the heart of a fish in the Gulf of Mexico, and of these hosts, only the bluefish was previously identified as a host for a species of Kudoa. Spores of the new species varied slightly in size among these hosts but were regarded as conspecific based on their nearly identical (99.9%) small-subunit (SSU) ribosomal DNA (rDNA) sequence. The new species differs both from the 4 nominal species of Kudoa reported from fishes in the Gulf of Mexico and from K. pericardialis, an allopatric species that infects the pericardial cavity, by the combination of having a large spore, a small polar capsule, and a polar filament with a single coil. The new species is morphologically and genetically most similar to K. shiomitsui, an allopatric species that infects the heart and pericardial cavity, but is distinguished from it based on a 4.2% difference in the SSU rDNA sequence. Heart lesions primarily were restricted to the vicinity of plasmodia and included a layer of fibrinous inflammation characterized by lymphocytes, macrophages, and granulomas as well as epithelioid encapsulations around plasmodia. Heavily infected hosts had melanin-like deposits and adipose cells beneath the epicardium, and the epicardium was discontinuous and apparently breached by plasmodia in some regions. Cardiac muscle, gill, liver, spleen, intestine, and kidney were normal.

The species recently described as Acanthostomum macroclemidis possesses the gonotyl in the form of a solid muscular pad uniquely diagnostic for species of Proctocaecum and is accordingly transferred to that genus. An artificial key to the 5 acanthostomine genera, as well as an updated phylogenetic hypothesis for the 10 known species of Proctocaecum, based on 11 characters and including 2 species described since the last phylogenetic analysis, are presented. The single most parsimonious phylogenetic tree with a consistency index of 87.5% suggests that Proctocaecum originated in Africa and spread to North America and South America before the breakup of Pangaea. As a result, the 2 North American and 1 South American species are most closely related to different African members of the genus. African and Indo-Pacific species inhabit crocodylids; hence, the occurrence of North American species in alligatorids and chelonians and a South American species in alligatorids are the result of host switches.

Falcaustra costaricae n. sp. from the intestines of the lizard Norops tropidolepis and F. heosemydis n. sp. from the large intestine of the turtle Heosemys depressa are described and illustrated. Falcaustra costaricae represents the 10th Neotropical species assigned to this genus and is distinguished from other Neotropical species by the distribution pattern of caudal papillae (10 preanal, 0 adanal, 12 postanal, and 1 median), length and width of spicules (510–561 μm long, 18–24 μm wide), and absence of pseudosucker. Falcaustra heosemydis represents the 29th Oriental species and is distinguished from other Oriental species by the distribution pattern of caudal papillae (10 preanal, 0 adanal, 12 postanal, and 1 median), length of spicules (790–890 μm), and absence of pseudosucker. Norops tropidolepis was found to harbor 3 species of Nematoda, F. costaricae, Rhabdias anolis, and acuariid larvae, and 2 species of Acanthocephala (centrorhynchid cystacanths and oligacanthorhynchid cystacanths).

Saccocoelium megasacculum n. sp. (Digenea: Haploporidae) was collected from the intestine of the mugilid fish, Liza carinatus (Cuvier and Valenciennes), in the Taiwan Strait. It is the first record of Saccocoelium in China. The parasite most closely resembles Saccocoelium obesum Looss, 1902 and Saccocoelium tensum Looss, 1902 in general morphology and body size, but it is easily distinguished from them in having a larger hermaphroditic sac in relation to body size; larger eggs; smaller pharynx, testis, ovary, and vitellaria; and a uterine seminal receptacle instead of a true seminal receptacle.

During a helminthological examination of marine fishes from south of the Minnan-Taiwan Bank Fishing Ground, Taiwan Strait, Fujian, China, a new cucullanid nematode, Dichelyne (Cucullanellus) jialaris n. sp., was removed from the intestine of the red seabream, Pagrus major (Temminck & Schlegel, 1834). The new species differs from its congeners mainly in the following characters: body size medium but with relative long spicules of 1.01 mm (0.97–1.06) in length or 20.0% (18.21– 21.8%) of the body length; proximal end of spicules somewhat expanded and distal end rounded; gubernaculum I-shaped, slightly narrow in the middle part, both ends rounded; both anterior and posterior cloaca lips round or oval, prominent and unequal in size. The anterior cloaca lip is at least 2 times larger than the posterior one. There is a conspicuous papilliform structure within the central of anterior and posterior cloacal lip. Vulva of female is not prominent, slightly postequatorial; distance from vulva to anterior end of body is 4.3 (3.0–5.5) mm or 58.0% (54.0–62.0%) of the body length. Considering the result of comparing the structure of so-called unpaired median papilla with the 10 pairs of caudal petiolated papillae in the body of the same individual, the papilliform structures are just a backstop for the cloacal lips, this new species represents the first record of a nematode of the Dichelyne, subgenus Cucullanellus in marine fishes of China Sea.

Phylogenetic relationships between Bothriocephalus species from freshwater and marine teleosts from different geographical regions were studied using internal transcribed spacer–2 and partial 18S ribosomal DNA. The analyses revealed a paraphyly of Bothriocephalus with respect to the genera Polyonchobothrium, Anantrum, and Clestobothrium. The freshwater species Bothriocephalus claviceps, B. acheilognathi, and Bothriocephalus sp. from Dorosoma petenense formed a well-supported monophyletic cluster, with Polyonchobothrium at its base. In contrast, the type species, B. scorpii, clustered within a distinct lineage formed by a heterogeneous assemblage of marine species, Clestobothrium crassiceps and Anantrum tortum, and the freshwater species B. cf. japonicus. This shows that the current morphology-based classification is unlikely to reflect the phylogenetic relationships within this group and will require a thorough revision.

Fish parasites of the Multivalvulida (Myxozoa, Myxosporea) are widespread and can be associated with mortality or poor flesh quality in their commercially important marine hosts. Traditional classifications divide members of this order into families based on spore valve and polar capsule numbers. Analyses of the small-subunit (SSU) ribosomal DNA (rDNA) sequences from all representative families in the order (Trilosporidae, Kudoidae, Pentacapsulidae, Hexacapsulidae, and Septemcapsulidae) indicate that a revision of the taxonomy and nomenclature is warranted. In our phylogenetic analysis of (SSU and large subunit) rDNA sequences, members of Pentacapsula, Hexacapsula, and Septemcapsula root within a clade of Kudoa species with Unicapsula (Trilosporidae) as an outlier to these genera. Therefore, we propose to synonymize Pentacapsulidae, Hexacapsulidae, and Septemcapsulidae with Kudoidae alter the diagnosis of Kudoidae and Kudoa to accommodate all marine myxozoan parasites having 4 or more shell valves and polar capsules.

Parabothriocephalus psenopsis n. sp. (Eucestoda: Pseudophyllidea) is described from the gastrointestinal tract of Psenopsis anomala caught off the coast of Xiamen, China. The new species most closely resembles, but differs from, Parabothriocephalus segmentatus in its possession of muscular globular appendages on the posterior margin of the proglottids, a limited proglottid number (9–13), and the shorter strobila (7.6–13.2 mm vs. 165 mm). In addition, the uterus of P. psenopsis is strongly coiled, whereas that of P. segmentatus is S shaped; P. segmentatus has a spherical expansion in the middle of the vagina, whereas that of P. psenopsis does not. Finally, P. psenopsis differs from Parabothriocephalus gracilis, Parabothriocephalus sagitticeps, and Parabothriocephalus macruri by the posterolateral expansion of the proglottids.

Two new species of habronematid nematodes are described in birds from the Area de Conservacion Guanacaste, Costa Rica. Torquatoides trogoni n. sp., in Trogon massena, can be distinguished from T. torquata, T. bengalensis, and T. crotophaga in lacking lateral alae. Among species lacking lateral alae, the new species differs from T. balanocephala in having 14 versus 8–10 cephalic cuticular plaques, 21–22 versus 13–17 pairs of preanal papillae, and a beak-shaped versus U-shaped gubernaculum. The new species differs from T. singhi in body length, in having 21–22 versus 10 pairs of precloacal papillae, longer spicules, and larger eggs. The new species differs from T. crotophaga, the only other species known from Central America, in lacking lateral alae, and having 14 versus 6 cephalic cuticular plaques, 21–22 versus 17 pairs of precloacal and 3 versus 2 pairs of postcloacal papillae, and a gubernaculum. Excisa ramphastina n. sp., in Ramphastos sulfuratus, can be distinguished from E. excisa, E. biloba, E. buckleyi, E. dentifera, and E. khalili in having 1 lateral ala versus none, cervical papillae anterior versus posterior to the nerve ring, and asymmetrical caudal alae. Excisa ramphastina is similar to E. curvata in having cervical papillae anterior to the nerve ring but differs in having 1 lateral ala versus none, asymmetrical caudal alae, an average spicule ratio of 1:4.4 versus 1:3.3, and 4 versus 2 pairs of sessile papillae. The new species differs from E. columbi in having 1 versus 2 lateral alae, in the length of the spicules, in having a different spicule ratio, and in the numbers of sessile papillae.

Adult and larval stages of Taenia solium cause 2 diseases in humans, i.e., taeniasis and cysticercosis, respectively. Diagnosis and treatment of taeniasis are the ultimate means to eliminate cysticercosis. A serological taeniasis diagnostic test has been developed for laboratory use. However, recombinant forms of the taeniasis diagnostic proteins are required to overcome the limited supply of native proteins and allow the development of a low-cost and field-applicable test with high sensitivity and specificity. Using 2-dimensional electrophoresis of T. solium excretory and secretory (TSES) products from hamster adult tapeworm in vitro cultures, we have identified 5 T. solium–specific protein spots, with molecular weights of 33 kDa (protein isoelectrofocusing point [pI]: 5.6, 5.3, 5.1) and 38 kDa (pI: 4.6, 4.5). Protein sequencing and molecular cloning of these proteins showed that although endowed with different pIs, the proteins with the same molecular weights shared the same protein backbone, named TSES33 and TSES38. Their full-length complementary DNAs encode proteins with 267 and 278 amino acids, respectively. TSES33 and TSES38 were expressed in a baculovirus system. Both recombinant proteins were recognized by a panel of taeniasis, but not cysticercosis patient serum samples, indicating that they can potentially replace the native proteins in the development of a more efficacious taeniasis diagnostic test.

Toxoplasma gondii is an important apicomplexan parasite of humans and other warm-blooded animals. Ponazuril is a triazine anticoccidial recently approved for use in horses in the United States. We determined that ponazuril significantly inhibited T. gondii tachyzoite production (P < 0.05) at 5.0, 1.0, or 0.1 μg/ml in African green monkey kidney cells. We used outbred female CD-1 mice to determine the efficacy of ponazuril in preventing and treating acute toxoplasmosis. Each mouse was subcutaneously infected with 1,000 tachyzoites of the RH strain of T. gondii. Mice were weighed daily, and ponazuril was administered orally in a suspension. Mice given 10 or 20 mg/kg body weight ponazuril 1 day before infection and then daily for 10 days were completely protected against acute toxoplasmosis. Relapse did not occur after prophylactic treatments were stopped. Toxoplasma gondii DNA could not be detected in the brains of these mice using polymerase chain reaction (PCR). One hundred percent of mice treated with 10 or 20 mg/kg ponazuril at 3 days after infection and then daily for 10 days were protected from fatal toxoplasmosis. Sixty percent of mice treated with 10 mg/kg ponazuril at 6 days after infection and 100% of mice treated with 20 mg/kg or 50 mg ponazuril 6 days after infection and then daily for 10 days were protected from fatal toxoplasmosis. Relapse did not occur after treatments were stopped. Toxoplasma gondii DNA was detected in the brains of some, but not all, of these mice using PCR. The results demonstrate that ponazuril is effective in preventing and treating toxoplasmosis in mice. It should be further investigated as a safe and effective treatment for this disease in animals.

Although Toxoplasma gondii infection has been found occasionally in cetaceans, little is known of the prevalence of antibodies to T. gondii in wild dolphins. Antibodies to T. gondii were determined in serum samples from 58 dolphins stranded in the Spanish Mediterranean coast. Modified agglutination test was used to determine T. gondii antibodies, and a titer of 1:25 was considered indicative of T. gondii infection. Antibodies to T. gondii were found in 4 of 36 striped dolphins (Stenella coeruleoalba), in 2 of 4 common dolphins (Delphinus delphis), in 4 of 7 bottlenose dolphins (Tursiops truncatus), and in 1 harbour porpoise (Phocoena phocoena). Antibodies were not found in 9 Risso's dolphins (Grampus griseus) and in 1 long-finned pilot whale (Globicephala melas) surveyed. The results indicate that T. gondii infection is frequent in at least 3 dolphin species from the Mediterranean Sea.

Black turtle (Chelonia mydas agassizii) carcasses, recovered as a result of incidental capture in Magdalena Bay, Mexico, revealed invasion by spirorchiid trematode eggs in liver, kidney, intestines, muscle, heart, pancreas, and duodenum. Seventy-five adult Learedius learedi Price, 1934, were recovered from the heart of 1 turtle. Most of the organs showed a mild or absent inflammatory response in histological sections, with the exception of a pancreatic–duodenal section that revealed severe lymphocyte and phagocyte infiltration associated with an infestation of more than 200 eggs. A linear formation of 35 eggs from the pancreas toward the intestinal lumen is described as resembling migration. This is among the first reports of a parasitic infection of L. learedi Price 1934, in C. m. agassizii in Mexico.

Adults of Trichuris skrjabini have been isolated from the cecum of caprine hosts (Capra hircus), Trichuris ovis and Trichuris globulosa from Ovis aries (sheep) and C. hircus (goats), and Trichuris leporis from Lepus europaeus (rabbits) in Spain. Genomic DNA was isolated and the ITS1–5.8S–ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced by polymerase chain reaction (PCR) techniques. The ITS1 of T. skrjabini, T. ovis, T. globulosa, and T. leporis was 495, 757, 757, and 536 nucleotides in length, respectively, and had G C contents of 59.6, 58.7, 58.7, and 60.8%, respectively. Intraindividual variation was detected in the ITS1 sequences of the 4 species. Furthermore, the 5.8S sequences of T. skrjabini, T. ovis, T. globulosa, and T. leporis were compared. A total of 157, 152, 153, and 157 nucleotides in length was observed in the 5.8S sequences of these 4 species, respectively. There were no sequence differences of ITS1 and 5.8S products between T. ovis and T. globulosa. Nevertheless, clear differences were detected between the ITS1 sequences of T. skrjabini, T. ovis, T. leporis, Trichuris muris, and T. arvicolae. The ITS2 fragment from the rDNA of T. skrjabini was sequenced. A comparative study of the ITS2 sequence of T. skrjabini with the previously published ITS2 sequence data of T. ovis, T. leporis, T. muris, and T. arvicolae suggested that the combined use of sequence data from both spacers would be useful in the molecular characterization of trichurid parasites.

A serological survey of Toxoplasma gondii infection among slaughtered pigs in the largest slaughterhouse located in Taoyuan County of northwestern Taiwan was conducted using the latex agglutination (LA) test during 1998. The overall seroprevalence of T. gondii infection was 28.8% (32/111) with LA titers of 1:32 (6, 18.8%), 1:64 (10, 31.2%), 1:128 (9, 28.1%), 1:256 (6, 18.8%), and 1:512 (1, 3.1%). No significant difference (P > 0.05) in seroprevalence between male (28.6%, 20/70) and female (29.7%, 12/41) slaughtered pigs was observed. A decreasing trend in the seroprevalence among slaughtered pigs examined in the same slaughterhouse was observed because of a lower seroprevalence (P < 0.05) than that (44.4%, 128/288) previously reported about 10 yr ago using the LA test. Nevertheless, it is important to avoid eating raw or undercooked pork in order to prevent the acquisition of T. gondii infection among people in Taiwan.

To determine how long spores of Encephalitozoon cuniculi, E. hellem, and E. intestinalis remain viable in seawater at environmental temperatures, culture-derived spores were stored in 10, 20, and 30 ppt artificial seawater at 10 and 20 C. At intervals of 1, 2, 4, 8, and 12 wk, spores were tested for infectivity in monolayer cultures of Madin Darby bovine kidney cells. Spores of E. hellem appeared the most robust, some remaining infectious in 30 ppt seawater at 10 C for 12 wk and in 30 ppt seawater at 20 C for 2 wk. Those of E. intestinalis were slightly less robust, remaining infectious in 30 ppt seawater at 10 and 20 C for 1 and 2 wk, respectively. Spores of E. cuniculi remained infectious in 10 ppt seawater at 10 and 20 C for 2 wk but not at higher salinities. These findings indicate that the spores of the 3 species of Encephalitozoon vary in their ability to remain viable when exposed to a conservative range of salinities and temperatures found in nature but, based strictly on salinity and temperature, can potentially remain infectious long enough to become widely dispersed in estuarine and coastal waters.

Ixodid ticks were collected and identified from 8 wild carnivore species in Phu Khieo Wildlife Sanctuary, northeastern Thailand. Six tick species belonging to 4 genera were recovered and identified from 132 individuals. These included Amblyomma testudinarium (n = 36), Haemaphysalis asiatica (n = 58), H. hystricis (n = 31), H. semermis (n = 3), Rhipicephalus haemaphysaloides (n = 3), and Ixodes granulatus (n = 1). Leopard cats (Prionailurus bengalensis) (n = 19) were infested with 4 tick species, whereas yellow-throated marten (Martes flavigula) (n = 4), clouded leopard (Neofelis nebulosa) (n = 2), and dhole (Cuon alpinus) (n = 1) were infested with 3 tick species, Asiatic golden cat (Catopuma temmincki) (n = 2) with 2 species, and marbled cat (Pardofelis marmorata), binturong (Arctictis binturong), and large Indian civet (Viverra zibetha) each infested with 1 species. This information contributes to the knowledge available on the ectoparasites of wild carnivores in Southeast Asia.

A male harbor seal (Phoca vitulina) was found moribund on the coast of New Jersey in January of 2003 and died a few hours later in the Marine Mammal Stranding Center. On necropsy, a single female Dioctophyme renale was recovered from the peritoneal cavity, and a tissue mass was found adjacent to the pelvic urethra and urinary bladder. Within this tissue mass were found D. renale ova. This is the first report of this nematode in the harbor seal and in a North American marine mammal.

The purpose of the present study was to characterize the intracellular distribution of a native Neospora caninum 56-kDa protein that is recognized by sera from N. caninum–infected dairy cattle. The complementary DNA coding for this protein was expressed in Escherichia coli as a polyHis fusion protein to which antiserum was prepared and used to localize the antigen in N. caninum tachyzoites and bradyzoites. By sodium dodecylsulfate–polyacrylamide gel electrophoresis and immunoblotting, antirecombinant Nc56 serum recognized a major 56-kDa protein and 2 minor (43 and 39 kDa) proteins of N. caninum tachyzoites. Antiserum to recombinant 56-kDa protein showed this antigen to be present in both N. caninum tachyzoites and bradyzoites/cysts as detected by immunofluorescence staining. Immunoelectron microscopy revealed the 56-kDa antigen to be present in the apical end of both tachyzoites and bradyzoites and possibly extracellularly secreted by tachyzoites.

A total of 65 largemouth bass, Micropterus salmoides, and 27 smallmouth bass, M. dolomieu, collected in April–September 2000 and April–July 2001 from Gull Lake, Michigan, were examined for acanthocephalans. Leptorhynchoides thecatus and Neoechinorhynchus cylindratus infected all the bass examined. Leptorhynchoides thecatus had the highest mean intensity (258.2 ± 185.4 in 2000 and 145.0 ± 61.0 in 2001) of the species infecting smallmouth bass. Although N. cylindratus had higher mean intensities (42.1 ± 37.9 in 2000 and 68.9 ± 70.5 in 2001) than did L. thecatus in largemouth bass, the values were not significantly different between bass species. The prevalence, mean intensity, and mean abundance of Pomphorhynchus bulbocolli in the bass species were below the values for the other acanthocephalan species. Leptorhynchoides thecatus and N. cylindratus are the most abundant intestinal helminths in bass from Gull Lake.

Thin-layer chromatography was used to analyze the free-pool amino acids of the digestive gland–gonad complex (DGG) of Biomphalaria glabrata infected with Echinostoma caproni and uninfected (control) snails. Qualitative analysis revealed the presence of histidine, lysine, serine, alanine, valine, and isoleucine or leucine in all samples. Quantitative analysis of lysine and valine gave mean weight percentages of 0.00699 ± 0.0022 and 0.00174 ± 0.00056, respectively, in the DGG of uninfected snails, and 0.00504 ± 0.0014 and 0.00254 ± 0.00033, respectively, in the DGG of infected snails. The differences in values between infected and uninfected snails were not statistically significant (Student's t-test, P > 0.05).

Whether the brown rat (Rattus norvegicus) is a reservoir of Trichinella spp. infection or merely an accidental host, which may be vector of Trichinella spp., continues to be debated. We estimated the prevalence of Trichinella sp. infection in brown rat populations and in domestic pigs in 2 villages in Croatia, where Trichinella sp. infection in pigs has been endemic in the past 10 yr. Trichinella spiralis larvae, identified by a multiplex polymerase chain reaction analyses, were the only species detected in both rats and pigs. In 2001 and 2002, 2,287 rats were collected on 60 farms with different levels of sanitation and with, or without, T. spiralis–infected pigs. The prevalence of infection in rats ranged from 0.2 to 10.7%. Infected rats were detected only on farms with T. spiralis–positive pigs and low sanitation or formerly with low sanitation (P = 0.007, Fisher's exact test), yet no infected rat was detected on farms with T. spiralis–negative pigs. The finding that no infected rat was found on farms with T. spiralis–negative pigs suggests that, in the investigated area, the brown rat is not a reservoir but only a victim of improper pig slaughtering.

Molecular methods were used to identify blood parasites frequently observed in blood smears of water pythons (Liasis fuscus) captured in our study area in the Northern Territory of Australia. A nested polymerase chain reaction (PCR) using primers amplifying the 18s ribosomal RNA (rRNA) nuclear gene resulted in a short PCR product (180 bp) matching this region in the genus Hepatozoon. However, because of the short sequence obtained, 2 new primers were designed based on 18s rRNA sequences of 3 Hepatozoon taxa available in GenBank. Using these primers, approximately 600 bp of the parasite's 18s rRNA gene was amplified successfully and sequenced from 2 water python samples. The new primers were used to investigate the prevalence of blood parasites in 100 pythons. In 25 of these samples we did not observe any blood parasites when examining stained slides. All the samples revealed a 600-bp PCR product, demonstrating that pythons in which we did not visually observe any parasites were infected by Hepatozoon spp. We also analyzed the nucleotide sequences of blood parasites in 4 other reptile taxa commonly encountered in our study area. The sequences obtained from water pythons and from 1 of these taxa were identical, suggesting that the parasite is capable of infecting hosts at different taxonomic levels.