In Lake Fjellfrøsvatn, northern Norway, the larval helminths Cyathocephalus truncatus and Cystidicola farionis use Gammarus lacustris as intermediate hosts and Arctic charr (Salvelinus alpinus) as final hosts. There was sampled 1,433 live G. lacustris from the lake and 1,964 G. lacustris from stomach contents of the charr. Prevalence of infection were, respectively, 0.49% and 3.72% for C. truncatus, and 0.21% and 0.20% for C. farionis. Usually, only 1 parasite was present in each host, and the 2 parasite species never co-occurred. Gammarus lacustris amphipods parasitized by C. truncatus were positively selected by the Arctic charr and were consumed approximately 8 times as often as were the unparasitized amphipods or the amphipods infected with C. farionis. This suggests that G. lacustris amphipods infected with C. truncatus larvae are more susceptible to predation than noninfected specimens, probably because of parasite-induced alterations in behavior or visibility. Alternatively, this could also be explained by selection toward the largest G. lacustris specimens observed, which are also the most frequently parasitized amphipods. However, the data show clearly that this was not a result of size-selective predation by the charr. In contrast, the presence of C. farionis did not increase the susceptibility to predation of its intermediate host. The discrepancy between the 2 helminth species supports the hypothesis that parasite-increased susceptibility to predation is related to the life history strategies of the parasites.

There is an increasing interest in identifying the parasite components involved in the maturation, development, and infectivity of intracellular protozoan parasites. In the present study, a heat shock protein (hsp) of the family of 70 kDa hsp (hsp70), which play important roles in the stage conversion and virulence of these parasites, was examined. Whereas hsp70 expression has been examined in Eimeria tenella within host tissues, in the present study, oocysts of E. tenella were used to investigate the expression of hsp70 during sporulation without interference from the host; hsp70 expression during excystation was induced by incubating sporulated oocysts under various experimental conditions to produce the stimuli necessary for sporozoites to become active and to excyst in vitro. Hsp70 was detected by immunohistochemical techniques; quantitative flow cytometric analysis was also been carried out using specific monoclonal antibodies against hsp70. Hsp70 was expressed during sporulation but was not found in sporulated oocysts after the completion of sporulation. Oocysts re-expressed hsp70 when excystation was induced. The presence of hsp70 prior to infection may preadapt the parasite for additional stress in the host and may be involved in the formation of sporozoites.

Late summer myxozoan infra- and component communities parasitizing 73 Notropis hudsonius at 5 sites on the St. Lawrence River upstream and downstream from the island of Montreal are described from study of histological sections of individual fish. Community membership included Myxobolus sp. A (intracellular in striated muscle fibers), Myxobolus sp. B (intracellular in striated muscle fibers), Myxobolus sp. C (brain), and Thelohanellus notatus (Mavor, 1916) Kudo, 1929 (loose connective tissue), all of which are histozoic myxobolids displaying strict tissue specificity for trophozoite development. Mean infracommunity richness in fish at the separate localities was estimated to be 0.4 ± 0.5 to 1.3 ± 0.7, with a maximum richness of 3 in any 1 fish. Component community richness in host samples was 2 to 4. It is argued that these are relatively high levels of diversity for freshwater fish parasites but that the values are probably conservative because of the study of only portions of individual fish. The percentage of fish infected with myxozoans of any species and infracommunity richness was significantly greater below the island of Montreal compared with above it. It is suspected that increased oligochaete populations at these sites, resulting from sewage-caused organic enrichment of sediments, may have accounted for the observed increased prevalence of infection.

Recently transformed elvers of Anguilla rostrata, entering the mouth of the East River in the Timber/Connaught Lake watershed (Nova Scotia) from the sea in mid-May, did not develop infections of the urinary bladder parasite Hoferellus gilsoni (Myxozoa) after nearly 3 mo in captivity. However, 40% of elvers collected from the mouth of the river in early June and 80% in late June exhibited patent infections by mid-August. It is speculated that given this initial level of infection, the summer run of elvers (estimated to be 208,000–963,000 individuals annually) represents a significant upstream dispersal of spores. Infections in yellow eel, which has a limited home range, probably contribute to spore dispersal within the home range and downstream through spore drift. Sporogenesis occurred in samples of yellow eel collected over an 8-mo period from April to November. Eel length was similar in 3 widely separated sampling locations in the watershed. Prevalences at these localities were 44, 36, and 57% in April and, by June, reached 76, 70, and 90%, respectively. Prevalences remained near the June levels until sampling ceased in November 1998. Studies on captive fish suggest that H. gilsoni can overwinter in the eel. Data from the study indicate that immediate infection of young elvers entering the system, effective spore dispersal, and a seasonally extended sporogenesis all contribute to the ubiquitous presence of the parasite in the eel population.

A total of 239 green frogs Rana clamitans, collected between June 3 and August 27, 1998 from 6 locations in southwestern Michigan, was examined for helminths. Of the 26 helminth taxa found, the larval cestode Mesocestoides sp. had the highest mean intensity, followed by the larval trematode Fibricola sp. Of the helminths that mature in frogs, Haematoloechus varioplexus had the highest prevalence and Gorgodera amplicava had the highest mean intensity. Frogs from 118th Pond had the highest species richness (20), mean helminth species richness (5.2), and mean helminth abundance (153.7). Frogs from Constantine East had the highest mean helminth species diversity (0.8778) and evenness (0.6033), followed by frogs from 118th Pond. In all comparisons of mean helminth community species richness, abundance, diversity, and evenness, adult frogs had significantly higher or higher values than did juveniles at each location. Jaccard's coefficients of similarity for the helminth communities for location pairs ranged from 0.545 to 0.823. Nine and 2 core helminth taxa occurred at the local and regional levels, respectively. The differences in several helminth community measures in green frogs among locations stress the importance of local ecological conditions on helminth community structure.

Here the use of repeated measure linear modeling to analyze experimental infections is demonstrated. This method of analysis makes full use of serial measurements on the same host throughout the course of an infection while taking into account correlations between measures within the same host. As an example, repeated measure linear modeling is used to analyze worm output from an experimental infection of Strongyloides ratti, a nematode parasite of rats. This analysis allows differences in the temporal dynamics of infection of two lines of S. ratti to be dissected.

Gill lesions associated with infections of Erpocotyle tiburonis (Brooks, 1934) (Monogenea: Hexabothriidae) on wild bonnethead sharks (Sphyrna tiburo (L., 1758) (Carcharhiniformes: Sphyrinidae)) were compared with those on aquarium-held ones using light and scanning electron microscopy. Uninfected gill filaments had slender, triangular, smooth-surfaced lamellae and interlamellar water channels that were approximately equal in size. Four wild sharks were each infected by 3–11 widely separated adult E. tiburonis, and 1 of these sharks hosted a juvenile specimen. Lamellae flanking or touching adult E. tiburonis were pushed aside or bent, but were otherwise identical to those of uninfected filaments. Two aquarium-held sharks were each infected by hundreds of juvenile and adult E. tiburonis. In these sharks, lamellae near juveniles were pushed apart or bent, but were otherwise normal, whereas a thick, ragged-surfaced layer of hyperplastic epithelium both filled interlamellar water channels and partially or completely covered lamellae near adults. Results of this study suggest that the intense infections of E. tiburonis were facilitated by captivity and caused severe hyperplastic lesions that ultimately led to the death of the sharks by reducing or blocking the respiratory water flow over lamellae and thus reducing the exchange of gases and ions across the lamellar epithelium. In contrast, the wild sharks were infected by fewer worms and exhibited relatively minor lesions.

Remote sensing on the basis of AVHRR (advanced very high resolution radiometer) satellite imagery was used, together with standard geostatistical methods (cokriging), to estimate the distribution of habitat suitability (HS) for the tick Boophilus microplus (Canestrini) in Central America. Most attention was paid to the expected limits of tick distribution as well as the temperature and vegetation values responsible for different HS zones and their variability within the year. The highest HS extended through wide areas of southeastern United States, much of Mexico, and other countries of Central America. Areas with higher HS had relatively stable temperatures (within 20–25 C) throughout the year, and had a NDVI (normalized derived vegetation index) of around 0.4. These areas need to be targeted to ensure acaricide usage at optimum intervals and to avoid the formation of areas with enzootic instability. A seasonal analysis of the climate trend through the study period (1982–1999) revealed a warming cycle, together with a rise in NDVI index values. The main consequence of this trend is the gradual increase in unsuitability in areas where the tick is already established, with the establishment of new foci in zones currently too cold to support tick populations. The cooling periods of 1–2 yr that were observed between warming cycles may also lead to enzootic instability when warm zones cool to within the suitability range. As the model is remotely sensed, a continuous update of the B. microplus distribution could be performed, assuring maximum efficiency in developing management strategies.

The host–parasite interface between 2 species of damselflies and 4 species of eugregarines was examined at the ultrastructural level. Nubenocephalus nebraskensis organisms attached to the host midgut epithelium by means of a sucker-like protomerite; the space between the epicytic folds and host epithelium was filled with electron-dense material interpreted to be adhesive in nature. Actinocephalus carrilynnae organisms attached by means of the epimerite, which had no epicytic folds, and by the fluted stalk with characteristic epicytic folds; host cell and parasite membranes appeared fused at some places on the epimerite. Hoplorhynchus acanthatholius organisms attached by means of an ovoid epimerite with backward-pointing digitations; the entire epimerite was embedded in a host cell, and host cell microvilli surrounded the stalk. Steganorhynchus dunwoodyi organisms attached by means of an ovoid stalk papilla enclosed in a retractable globular sheath; the papilla was covered with epicytic folds, but the sheath was not, and the sheath had a single membrane, whereas the epicytic folds had 2 or 3 membranes. The entire apparatus was inserted between epithelial cells, and the sheath was highly folded at its surface. The ultrastructural observations suggest that actinocephalid gregarines have evolved 2 general strategies for attaching to the host epithelium, that is, suckerlike protomerites, as in the case of N. nebraskensis, and deeply embedded epimerites inserted within or between host cells, as in the other species studied.

The present work deals with optimization of excystation of Cryptosporidium parvum oocysts and the infection process of tissue culture cells by the parasite. It was shown that presence of the bile salt sodium taurocholate in the incubation medium expedited excystation of the tested GCH1 isolate and enhanced it, as compared with bleaching of the oocysts. This bile salt had no effect on the viability of tissue culture cell lines MDBK and HCT-8 at the tested concentration of 0.375% for up to 2 hr of coincubation. Infection studies conducted on tissue culture cells showed higher infection rates in the presence of sodium taurocholate than with bleached oocysts in the absence of this bile salt. It may be concluded that, at least as regards the GCH1 strain of C. parvum, the whole infection process can be performed in the presence of sodium taurocholate, and does not require separation and cleaning of the excysted sporozoites before their application to tissue culture cells.

In European freshwater, cyprinid fish may be heavily infected by plerocercoids of the pseudophyllidea cestode Ligula intestinalis (L.). During their development, these parasites grow rapidly to a large size in the fish's body cavity, characteristically distending the abdomen. In this study, the influence of this tapeworm on roach (Rutilus rutilus L.) morphology was analyzed. Forty-five infected and 45 uninfected roach were collected from the Lavernose-Lacasse gravel pit in Toulouse, south western France and examined for 40 morphological measurements to study phenotypic modification of the body and 14 bilateral characters for an analysis of asymmetry. Results indicate that the degree of bilateral asymmetry does not change between infected and uninfected roach, despite the strong host-morphological modifications such as deformation of the abdomen, fin displacements at the level of the tail, and sagging of the vertebral column. The intensity of abdominal distension and fish morphology changes depends on the total parasite biomass present. Differences were observed in morphology at different levels of infection, which relate to established effects of L. intestinalis on the physiology and behavior of intermediate hosts. These morphological changes induced by the parasite could increase trophic transmission to the definitive avian hosts.

The Schistosoma mansoni mitochondrial genome contains tandemly arrayed copies of a 62-base repeat motif. The tandem array is highly polymorphic with respect to number of repeats and commonly exhibits heteroplasmy. This study shows that a very high rate of mutation rapidly produces new repeat lengths (new haplotypes) for this mitochondrial variable number tandem repeat. A maternal inheritance pattern is also demonstrated for this repeat sequence, while the high mutation rate causes some offspring to exhibit nonmaternal haplotypes. Frequent generation of new haplotypes can be observed within samples of clonal cohorts taken from monomiracidial snail infections. These same clonal cercarial groups, when crossed, produce F₁ generations that exhibit the maternal set of haplotypes, across all individuals, with the frequent addition of new mutant haplotypes. In each of 2 crosses, a subset of the recently arisen haplotypes match paternal haplotypes by chance (30.4% and 18.8%), thus giving the false appearance of partial paternal inheritance of mitochondria.

Protein expression, characterized in Western blots and gelatinolytic activity, of cruzipain (Cr), the major Trypanosoma cruzi cysteine proteinase, was compared among 3 attenuated T. cruzi strains (TUL 0, TCC, and Y null) and their virulent counterparts (TUL 2, Tulahuen, and Y). All attenuated strains displayed a weaker gelatinolytic activity as compared with their virulent counterparts. The electrophoretic mobility and immunological reactivity revealed quantitative and qualitative differences, with the attenuated parasites showing bands of less density in all strains and lower mobility in 2 of them, as compared with the virulent strains. Sequence analysis of 1 Cr gene in the Tulahuen and TCC strains indicated 37/1404 base pair substitutions, corresponding to 20 amino acid changes in the attenuated strain. A similar comparative analysis of 1 Cr gene between Y and Y null strains showed 13/1404 base pair substitutions, corresponding to 8 amino acid changes in the attenuated strain. Although enough variability exists in the Cr gene to allow for less- or nonfunctional isoforms of the protein, further clones should be analyzed to establish whether attenuation is regularly associated with specific sequence changes of this enzyme.

The effect of oxidants and the anti-inflammatory steroid dexamethasone on the attachment and internalization of virulent and avirulent Leishmania donovani promastigotes by the macrophage mannosyl fucosyl receptor was examined. Oxidants and dexamethasone are known to down- and upregulate the expression of the mannose receptor. Macrophages, when treated with 500 μM H₂O₂ at 37 C for 30 min, stimulate about 45% inhibition in uptake of an avirulent strain (UR6), and 30 and 25% inhibition for virulent strains AG-83 and GE-I, respectively. Treatment of macrophages with dexamethasone for 20 hr resulted in a stimulation in uptake of the parasite. When UR6 was used, a 3-fold increase in uptake was observed compared with the controls. Parasite uptake was also inhibited by the H₂O₂-generating system, glucose/glucose oxidase; inhibition was blocked by catalase. Treatment of macrophages either with H₂O₂ or dexamethasone did not affect the binding of the advanced glycosylation end product–bovine serum albumin (AGE-BSA), the ligand for AGE receptor of macrophages. Similarly, indirect evidence also shows that both types 1 and 3 complement receptors (CR1, CR3) are not affected by these treatments, indicating that, besides the mannosyl fucosyl receptor, other receptors are minimally altered in the identified condition. These results suggest that the up- and downregulation of the mannose receptor of macrophages may play a role in affecting L. donovani infection.

A combination of molecular sieving chromatography and 2-step preparative isoelectric focusing showed that native Fh12, a fatty acid–binding protein isolated from Fasciola hepatica adult worms, is a protein complex of at least 8 isoforms with identical molecular mass but different isoelectric points. Using enzyme-linked immunosorbent assay (ELISA) and inhibition ELISA assays, immunological differences were observed between native (nFh12) and a recombinant molecule denoted rFh15 that was obtained after screening a cDNA library from F. hepatica adult worms with an anti-Fh12 monospecific polyclonal antibody. It was confirmed that in infected rabbits, antibodies to nFh12 appear by the second week postinfection, whereas antibodies to rFh15 appear much later, by 6 wk postinfection. Four acidic forms (Fh12_1–4) showed more immunological identity with rFh15 than with nFh12, based on the observation that they inhibited ELISA activity by nearly 50% when they were added to the anti-rFh15 polyclonal antibody at 20 μg/ml of protein concentration. Moreover, the Fh12_1–4 isoforms were poorly reactive with sera from rabbits 2–4 wk postinfection. However, the 2 acidic forms, denoted Fh12₅ and Fh12₆, and the neutral/basic forms, denoted Fh12₇ and Fh12₈, showed more immunological identity with the native nFh12 molecule than with the recombinant rFh15 because they were highly reactive with sera of rabbits with early 2-wk F. hepatica infection and inhibited ELISA activity nearly 50% when they were quantitatively added to the anti-nFh12 polyclonal antibody. These results suggest that rFh15 could be one of the acidic forms of nFh12, and that it, in fact, may be one of the less immunogenic or immunoprotective members, or both, of the nFh12 protein complex.

The effect of zinc supplementation on Taenia crassiceps murine cysticercosis was studied in susceptible BALB/cAnN mice. Female offspring of mice supplemented with high zinc throughout gestation and lactation were intraperitoneally infected with T. crassiceps cysticerci. Offspring from nonsupplemented mothers were used as controls. Significantly fewer parasites were recovered from zinc–supplemented mice (Zsm) 30 days after infection. Increased resistance was not related to the IgG antibody response. At early stages of infection, T cells from Zsm proliferated to T. crassiceps antigens, whereas cells from control mice did not respond. Infection caused in both groups a decrease in CD3 cell percentages, which was more pronounced in the controls, and paralleled by a decrease in CD8 cells; CD3 and CD8 percentages returned to normal levels at later stages of infection. In contrast, the CD4 subpopulation only decreased in control mice. Intracellular cytokine determinations indicate that zinc supplementation favored a stronger and persistent type-1 T cell response in cysticerci-infected mice, which probably participates in the observed increased resistance.

The cellular immune responses of cattle immunized with antigens of Tritrichomonas foetus were investigated. Subcutaneous injections of antigen preparations primed bovine peripheral blood mononuclear cells (PBMC) by 30 days of immunization as demonstrated by antigen-specific proliferation and by cytokine production upon antigen challenge of PBMC. Antigen-specific T-cells derived from PBMC responded by production of interferon (IFN)-γ message detected by reverse transcriptase polymerase chain reaction, secreted IFN-γ detected by enzyme-linked immunosorbent assay, and intracellular IFN-γ detected by flow cytometry. Phenotypic analysis of PBMC responding in vitro to parasite antigen demonstrated a shift from a mixed CD4 , CD8 , γδ , to predominantly CD4 , CD8⁻, γδ⁻ phenotype in the Tf190-primed PBMC. In conclusion, systemic immunization of cattle with parasite antigen results in priming of bovine T-cells that are antigen specific and can produce an anamnestic IFN-γ response to subsequent stimulation with antigens of T. foetus.

Freshwater nematomorphs, or gordiids, are parasitic as larvae, but free-living in aquatic environments as adults. Studies based on the collection of adults have reported gordiids to be widespread, but discontinuous in distribution. However, a relatively short adult life span and unknown life history make the detection of adults difficult. An alternative approach to investigate gordiid distribution is to use cysts. Of all paratenic hosts, snails were chosen because they lacked internal defense reactions to the cysts and become easily infected. Here, it is reported that the occurrence of gordiids on the basis of the cyst stage is much more common than previously reported, thus altering the perception of how common these worms are. Using this modified survey procedure, gordiid cysts were found at 70% of sites examined, in an area where extensive sampling over 3 yr yielded adults only at a single site. Of 1,000 snails dissected, 395 were infected with gordiids (intensity range: 1–465). Furthermore, different types of human-modified landscapes did not affect gordiid distribution, suggesting that as urban and suburban areas sprawl, human encounters or pseudoparasitism with nematomorphs may increase. The results of this study indicate that use of organismal-specific sampling techniques can be critical in studies of parasite distribution and biodiversity.

Aspects of the migratory life cycle and pathogenesis of Elaphostrongylus alces were studied in 7 randomly selected moose calves and 7 yearlings killed during August to November. One calf and 1 yearling were uninfected. The 6 infected calves had recent infections, whereas the 6 yearlings showed older infections from the summer of the previous year. The 2 calves killed in September had a total of 26 adult E. alces in the epidural space of the caudal vertebral canal and none in the skeletal muscles, whereas the remaining calves killed 1 to 2 mo later had 25 nematodes in the caudal and cranial vertebral canal and 7 in the skeletal muscles. The yearlings had a total of 101 adult E. alces in the skeletal muscles and 2 in the vertebral canal. There were no findings indicating involvement of the central nervous system in the life cycle of E. alces. Our findings suggest that E. alces migrates directly from the gut to the epidural space of the caudal vertebral canal where development to the adult stage takes place. During development, the nematode produces inflammation of the epidural tissue and spinal nerves. Development in the caudal vertebral canal is followed by some anterior dispersion of nematodes along the canal, and migration into skeletal muscles. Here the nematodes seem to live in reproductive pairs and groups. The predilection site for E. alces in moose is the muscles of the thigh.

Trypanosome infections identified by polymerase chain reaction on field-caught tsetse flies from various locations were analyzed with respect to factors intrinsic and extrinsic to the trypanosome–tsetse association. These factors were then simultaneously analyzed using artificial neural networks (ANNs) and the important factors were identified to predict and explain the presence of trypanosomes in tsetse. Among 4 trypanosome subgroups (Trypanosoma brucei s.l., T. congolense of the ‘savannah’ and of the ‘riverine-forest’ types, and T. simiae), the presence of the 2 types of T. congolense was predictable in more than 80% of cases, suggesting that the model incorporated some of the key variables. These 2 types of T. congolense were significantly associated in tsetse. Among all the examined factors, it was the presence of T. congolense savannah type that best explained the presence of T. congolense riverine forest type. One possible biological mechanism would be ‘hitchhiking,’ as previously suspected for other parasites. The model could be improved by adding other important variables to the trypanosome tsetse associations.

Tritrichomonas foetus is a common, sexually transmitted, protozoan parasite of cattle. It has an essential requirement for iron, which it obtains from host lactoferrin. However, specific lactoferrin-binding protein receptors have not yet been identified in T. foetus. To differentiate specific and nonspecific binding of lactoferrin, lactoferrin affinity chromatography and Western blotting was used to identify metabolically or surface-labeled T. foetus lactoferrin-binding proteins. Bovine lactoferrin was shown to bind more efficiently than human lactoferrin, and each of these bound much better than bovine transferrin. This is relevant because T. foetus is both species-specific and only infects the mucosal surface of the reproductive tract, which has little transferrin. Whereas the majority of lactoferrin binding was specific, competitive inhibition studies showed that nonspecific, charge-related binding of lactoferrin to T. foetus may also be involved. In the presence of bovine cervical mucus, binding of lactoferrin to T. foetus was diminished, suggesting that mucus has an effect on lactoferrin binding. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis of surface biotinylated proteins affinity-purified on lactoferrin–Sepharose showed biotinylated bands at M_r values of 22, 49, 55, 72, and 155 kDa. Because lactoferrin-binding proteins may be susceptible to digestion by T. foetus extracellular cysteine proteinases, it is suspected that the 155-kDa protein is the specific lactoferrin-binding protein and that the lower-M_r lactoferrin-binding molecules may be fragmentation products that contain the lactoferrin-binding site; however, other interpretations are clearly feasible. It is possible that there may be multiple proteins or multimers of the same protein. In summary, the data showed that binding of lactoferrin to T. foetus may be regulated by an interplay of specific receptor interactions as well as by hydrophobic and charge-related interactions.

Lipophosphoglycan (LPG), a major surface molecule from Leishmania donovani, stimulated ornithine decarboxylase (ODC) activity in macrophages in a dose- and time-dependent manner. LPG stimulated the rapid increase in ODC activity within 30 min after exposure, suggesting that the interaction of LPG with its receptor stimulated a specific signal transduction pathway. However, LPG-induced ODC activity was a transient event because 3 hr after exposure to LPG, no stimulation of ODC activity was detectable. ODC activity appeared to be coupled to the activation of protein kinase C (PKC) in macrophages, as activators of PKC caused a rapid increase in the ODC activity. Macrophages pretreated with LPG for 1 hr became unresponsive to subsequent stimulation by the PKC activators 1-oleoyl-2-acetyl-glycerol and the calcium ionophore A23187. In contrast, the ability of macrophages to express ODC activity in response to the cyclic AMP analogue dibutyryl cyclic AMP was not impaired by LPG.

Fecal samples from 11 colubrid snakes, representing 10 species, collected in Ecuador during October 1994 were examined for coccidian parasites. Feces of 4 individuals, representing 4 host species, contained coccidian oocysts. Three species of Eimeria and 2 species of Isospora were observed and are described here as new. Oocysts of both Eimeria and Isospora were found in the feces of a slug-eating snake, Dipsas vermiculata. Sporulated oocysts of the Eimeria sp. are spheroid to subspheroid, 16.7 by 16.6 μm (14–18 by 14–18 μm) and those of the Isospora sp. are spheroid and 15.0 μm (13–18 μm) in diameter. Imantodes cenchoa, the common bluntheaded treesnake, was infected with a species of Eimeria. These sporulated oocysts are ellipsoid, 23.3 by 16.2 μm (25–21 by 15–17 μm). Sporulated eimerian oocysts from Leptodeira annulata, the southern cat-eyed snake, are subspheroid, 22.5 by 18.8 μm (19–26 by 17–21 μm). Feces of a juvenile Imantodes lentiferus, the bluntheaded vine snake, contained ovoid to ellipsoid isosporan oocysts, which measured 21.6 by 15.0 μm (20–23 by 14–16 μm) when sporulated.

Falcaustra washingtonensis n. sp. (Nematoda: Kathlaniidae) from the intestine of the salamander Ambystoma tigrinum melanostictum is described and illustrated. Falcaustra washingtonensis n. sp. represents the 14th nearctic species assigned to this genus and is distinguished from other nearctic species by the distribution pattern of caudal papillae and number of posterior muscle groups of the male: 10 pairs of sessile caudal papillae—3 pairs precloacal, 1 pair adcloacal, 6 pairs postcloacal—and 5 posterior muscle groups.

Sarcocysts were found in the skeletal muscles of road-killed and live-trapped opossums collected in north central Florida. Sarcocysts were spindle-shaped and macroscopic and had an average measurement of 3.8 mm by 154.6 μm. Sarcocysts were only observed in skeletal muscle. Sarcocysts have invaginations throughout the sarcocyst wall, which is approximately 1 μm thick. Protrusions on the sarcocyst wall are stumpy and digitlike and contain fibrillar elements that extend from the interior portion of the cyst wall through the villi. A new name, Sarcocystis greineri, is proposed for this species.

Heliconema minnanensis n. sp. is described from the Chiloscyllium plagiosum south of the Minnan-Taiwan Bank Fishing Ground, Taiwan Strait. It is characterized by having the following combination of features. There are no longitudinal ridges in the posterior part of the body; the male has 4 pairs of precloacal papillae and 6–7 pairs of postcloacal papillae. The right spicule averages 0.2 mm long (1.1% of body length), and the left spicule averages 1.2 mm long (5.6% of body length), making an average spicule ratio of 1:5.2. The female has a vulva located on the left side of a thick membrane and 56–65% of the body length from the anterior end. Raphidascaris trichiuri (Yin and Zhang, 1983) comb. n., known previously from males only, is transferred from Cloeoascaris and redescribed, including material of both sexes from Muraenesox cinereus from the type locality.

In the course of a revision of species of Haemonchus Cobb, 1898 (Nematoda), commonly referred to as large stomach worms and significant pathogens of ruminants, a new species was discovered in the grey rhebuck Pelea capreolus, and the bontebok Damaliscus pygarthus, in South Africa. The new species, Haemonchus horaki, was previously reported as a long-spicule form of H. contortus (Rudolphi, 1803) Ransom, 1911. The new species, compared with H. contortus, can be distinguished by significantly longer spicules (555–615 μm vs. 383–475 μm); a synlophe with fewer ridges (26 vs. 30 in the region of the posterior part of the esophagus) that extend more posteriorly (within 1 mm of the copulatory bursa in males and postvulvar in females, vs. 2/3 to 3/4 of prebursal and prevulvar lengths); and an asymmetrical dorsal lobe with a long dorsal ray divided for more than half of its length, forming 2 branches of unequal length (vs. a dorsal ray divided for less than half of its length and forming 2 equal branches in H. contortus).

Fecal samples from 10 pygmy chameleons, Rampholeon temporalis (Matschie, 1892), an endemic species of the Usambara Mountains in northeastern Tanzania, were examined for coccidian parasites. Two (20%) chameleons were found to be passing oocysts of Eimeria Schneider. Comparison with other species of Eimeria indicates that the coccidian found represents a new species. Sporulated oocysts of Eimeria hajeki n. sp. are oval, 30.2 (29–31) by 23.5 (22–25) μm, with a shape index (length/width) of 1.3 (1.2–1.4) and a 2-μm-thick rough, bilayered wall. Micropyle and polar granule are absent. Sporocysts are oval to rhomboidal, 10.8 (9–11.5) by 8.8 (7.5–10) μm, with a shape index of 1.2 (1.15–1.3) and a wall composed of 2 valves joined by a suture.

A new nematode belonging to the Cystidicolidae is described. Pseudascarophis genypteri n. sp. was found in the intestine of the red ling Genypterus chilensis, sampled off Talcahuano, Chile. It is distinguished from Ascarophis species mainly by the absence of cephalic papillae and of submedial and medial labia. The other species in the genus, Pseudascarophis kyphosi Ko, Margolis, and Machida, 1985, found in Kyphosus cinerascens, and P. tropica (Solov'eva, 1996), found in Parupeneus chrysopleuron, differed principally from P. genypteri in the form of pseudolabia, number of pre- and postanal papillae, and length of spicules.

Margotrema guillerminae n. sp. is described from the cyprinid Hybopsis calientis and the goodeid Alloophorus robustus from Lake Zacapu, Michoacán State, in the Mesa Central of Mexico. The new species is easily distinguished from the only other species of the genus, M. bravoae, by having short ceca, extending posteriad to reach the region of the anterior testis, vitelline follicles that are mainly lateral to the ceca but with some intercecal (particularly at the ovarian region), and a seminal receptacle that is located immediately posterior to the ovary. Previous records of Urorchiinae gen. sp. and Margotrema sp. from Alloophorus robustus from Lake Pátzcuaro refer to M. bravoae, which appears to be specific to members of the Goodeidae.

Buckarootrema goodmani n. g., n. sp. is described from the small intestine of the Murray turtle, Emydura macquarii (Gray, 1830), from the vicinity of Warwick, Queensland, Australia. The distinctive taxonomic features include the vitellarium, which consists of 2 compact masses directly anterior to and occasionally overlapping the testes; the uterus with extensive pre- and postovarian coils; intestinal ceca with small, medial diverticula that terminate anterior to or at the anterior margin of the testes; a comma-shaped cirrus sac with both internal and external seminal vesicles. Phylogenetic systematic analysis of the genera of the Pronocephalidae including Buckarootrema and Notopronocephalus, the only other genus of pronocephalids reported from Australian freshwater turtles, indicates that Buckarootrema is the sister taxon of Neopronocephalus and Notopronocephalus is the sister group of the rest of the Pronocephalinae.

Partial plastid 23S and nuclear 18S rDNA genes were amplified and sequenced from 2 morphologically similar Eimeria species, E. antrozoi from a bat (Antrozous pallidus) and E. arizonensis from deer mice (Peromyscus spp.), as well as some other Eimeria species from bats and rodents. The phylogenetic trees clearly separated E. antrozoi from E. arizonensis. The phylogenies based on plastid 23S rDNA data and combined data of both plastid and nuclear genes grouped 2 bat Eimeria and 3 morphologically similar Eimeria species from rodents into 2 separate clades with high bootstrap support (100%, 3 rodent Eimeria species; 72–97%, 2 bat Eimeria species), which supports E. antrozoi as a valid species. The rodent Eimeria species did not form a monophyletic group. The 2 bat Eimeria species formed a clade with the 3 morphologically similar rodent Eimeria species (E. arizonensis, E. albigulae, E. onychomysis, all from cricetid rodents) with 100% bootstrap support, whereas 2 other rodent Eimeria species (E. nieschulzi, E. falciformis, from murid rodents) formed a separate clade with 100% bootstrap support. This suggests that the 2 Eimeria species from bats might be derived from rodent Eimeria species and may have arisen as a result of lateral host transfer between rodent and bat hosts.

Cryptosporidiosis has been traced to drinking contaminated surface water, which was either not treated or was ineffectively treated. Testing to detect Cryptosporidium parvum in surface water has been suggested to help prevent future outbreaks. In the present study, the same sample collection and filtration methods were used to compared sample processing and detection steps from 4 testing methods: a modified information collection rule (ICR) method and method 1623 (both developed by the U.S. Environmental Protection Agency), a flow cytometric method, and a solid-phase cytometric method. All of these methods use fluorescent antibody staining, which is only a presumptive indication of the presence of this parasite. Confirmation requires another assay. Methods were evaluated for both presumptive and confirmed detection. Solid-phase cytometry had the highest presumptive and confirmed detection rates. Flow cytometry had the next highest presumptive detection rate in reagent water but was third in spiked surface and tap waters, with no confirmation procedure. The ICR method had the third highest presumptive detection rate in reagent water and the second highest in spiked surface and tap waters but failed to confirm any oocysts. Method 1623 had significantly lower presumptive detection than any other method and a significantly lower confirmation rate than the solid-phase cytometry method.

The effects of in vitro treatment of cercariae, schistosomula, and adult worms of Schistosoma mansoni with 4 hypothalamic–pituitary–adrenal (HPA) axis hormones are described. Dehydroepiandrosterone (DHEA) had the strongest effect on viability. Cercariae were more susceptible to this hormone than schistosomula and adults. Mechanically transformed schistosomula showed 100% mortality (determined microscopically by progressive internal disorganization, development of lucent areas in the cytoplasm, and progressive loss of motility) after 48 hr, whereas physiologically induced schistosomula were more resistant, maintaining viability for up to 5 days of exposure. Males were considerably less sensitive than females to the lethal action of DHEA. When adult worms were paired, DHEA lethality was markedly reduced, with viability beginning to decrease only after 4 days in culture. Cortisol reduced the viability of each of the stages tested about equally. Corticotropin-releasing hormone (CRH) and adrenocorticotropin (ACTH) did not affect the viability of any stage. DHEA and cortisol significantly inhibited in vitro oviposition, whereas CRH and ACTH did not. DHEA and cortisol exerted their effects on schistosome viability and oviposition in a concentration-dependent manner. These results suggest possible new avenues for the control of schistosomiasis.

An immunochromatographic card test (ICT) that uses fingerprick whole blood instead of serum for diagnosis of bancroftian filariasis has recently been developed. The card test was validated in the field in Kenya by comparing its sensitivity to the combined sensitivity of Knott's concentration and counting chamber methods. A total of 102 (14.6%) and 117 (16.7%) persons was found to be microfilaremic by Knott's concentration and counting chamber methods, respectively. The geometric mean intensities (GMI) were 74.6 microfilariae (mf)/ml and 256.5 mf/ml by Knott's concentration and counting chamber methods, respectively. All infected individuals detected by both Knott's concentration and counting chamber methods were also antigen positive by the ICT filariasis card test (100% sensitivity). Further, of 97 parasitologically amicrofilaremic persons, 24 (24.7%) were antigen positive by the ICT. The overall prevalence of antigenemia was 37.3%. Of 100 nonendemic area control persons, none was found to be filarial antigen positive (100% specificity). The results show that the new version of the ICT filariasis card test is a simple, sensitive, specific, and rapid test that is convenient in field settings.

The dominant proteins released by Ascaris suum during development in vitro from the L3 to L4 stage were identified as collagenous cuticular proteins by sequence analysis and susceptibility to digestion by collagenase. Under reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), the collagen proteins separated into 3 groups with molecular weights estimated at 32 kDa, 54–60 kDa, and 71–91 kDa. The 32-kDa protein represents monomeric collagen; the 54–60- and 71–91-kDa components represent dimeric and trimeric forms, respectively, polymerized by nonreducible cross-links. Furthermore, the release of these forms of collagen was developmentally regulated, as exemplified by a sequential temporal progression from monomeric to dimeric to trimeric forms in association with the in vitro transition from L3 to L4. The data suggest that collagen released in vitro during development of A. suum L3 to L4 reflects the increased translation of collagen gene products and their initial assembly into higher molecular weight molecules associated with the synthesis of the L4 cuticle. A biotinylated dipeptidyl fluoromethylketone cysteine protease inhibitor (Bio-phe-ala-FMK) bound specifically to the 32-kDa collagen and, to a lesser extent, to a 30-kDa protein; binding was dependent on the presence of dithiothreitol (DTT) and was prevented by iodoacetamide. Because cysteine residues play an essential role in the initial assembly of the collagen monomers into the higher molecular weight oligomers present in the mature nematode cuticle, inhibition of molting of A. suum L3 to L4 by the cysteine protease inhibitor Z-phe-ala-FMK might be due to its binding to thiol groups of collagen monomers during a critical phase of collagen assembly. Prevention of cystine cross-links during this critical period of cuticle assembly by peptide-FMK inhibitors may represent a potential control mechanism having a novel mechanism of action.

Nodulisporic acid A (NSA) has been shown previously to be safe in dogs and to deliver >90% flea control for 4 days following a single oral administration. Three newly prepared nodulisporamide derivatives were subsequently identified from an artificial membrane flea feeding system as exhibiting potency substantially greater than NSA. To determine if they have superior in vivo activity, these 3 nodulisporamides, as well as NSA, were evaluated in dogs at 15 mg/kg/os. Parasite challenges were made by placing 100 live Ctenocephalides felis fleas onto the dorsum of dogs every 48 hr and examining efficacy at each of those intervals over a 22-day period. Results showed that NSA produced >90% efficacy at day 2 and 81% efficacy at day 4, and its residual flea killing fell to ∼50% by day 6 posttreatment. All dogs treated with the 3 new experimental nodulisporamides were 100% protected from flea challenges to day 8 posttreatment, and 2 of the compounds continued to produce >90% residual activity to 2 wk posttreatment. Pharmacokinetic analysis showed that plasma profiles and half-lives of NSA and these 3 new compounds correlated closely with flea efficacy. These results demonstrate that specific substitutions to the pharmacophore of NSA can substantially increase the duration of activity against fleas.

The indirect immunofluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were compared with blood culture for the detection of Trypanosoma cruzi infection in 83 raccoons (Procyon lotor) trapped in 4 counties of southeast Georgia. Both IFAT and ELISA detected 24 of 25 culture-positive samples (96% sensitivity). Cultures from 25 raccoons (30%) were positive for epimastigotes, whereas a total of 50 raccoons (60%) was seropositive by either the IFAT or ELISA. Forty-five of 83 serum samples (54%) were positive for anti-T. cruzi antibodies with the ELISA, and 47 were IFAT positive (57%). Forty-two of the 50 seropositive raccoons (84%) were seropositive by both tests. Endpoint titers of IFAT-positive samples were determined by testing doubling dilutions from 1:40 to 1:1280. High titers of 640 and 320 were observed for 4 raccoons trapped in 1 county (St. Catherines Island, Liberty County) and titers of 160 for 1–2 raccoons from each of the 4 counties sampled. IFAT titers and ELISA optical density values were positively correlated. Both serological tests have a high sensitivity and should be excellent tools for studying the prevalence of T. cruzi in wildlife populations.

Among the large cells located in the posterior of Echinostoma caproni and E. paraensei miracidia are secretory cells, germinal cells (GC), and undifferentiated cells. Secretory cells do not give rise to progeny, whereas GC do. Undifferentiated cells develop into GC that can also divide to produce embryos. Cleavage of GC of E. caproni occurs only after the parasite has entered the snail host and develops into a sporocyst. With E. paraensei, GC are larger than noted for E. caproni, and in 3 of 23 miracidia examined, germinal cell cleavage had occurred in the miracidium such that an embryo containing 20–25 blastomeres was present. Observations on the germinal elements of miracidia help to explain previous results showing that (1) E. paraensei sporocysts release mother rediae a few days earlier than do sporocysts of E. caproni, and that (2) a single mother redia is produced ahead of all others by sporocysts of E. paraensei, but not by sporocysts of E. caproni. The present study adds support to the concept that E. caproni and E. paraensei are distinct species.

The prevalence, abundance, and developmental status of the digenetic trematode Proctoeces lintoni Siddiqui et Cable 1960 were compared in 3 species of keyhole limpets Fissurella. A total of 197 limpets was collected at Caleta Chome, south-central Chile. Fissurella picta and F. costata had the highest prevalence of infection, whereas F. picta showed the greatest abundance of parasites, which increased with host shell length. However, the frequency of P. lintoni specimens with eggs in the uterus was greatest in F. costata. These results suggest that an increased rate of development of a parasite in the intermediate host may shorten the residence time necessary for maturation in the final host. Thus, faster development of the parasite in F. costata suggests the possibility that the parasites transmitted through this host species have shorter maturation times in clingfishes than individuals transmitted via other limpet species.

Recent successes in culturing intramolluscan larval stages of Schistosoma mansoni have relied on synxenic culture with a cell line (Bge) developed from embryos of a molluscan host Biomphalaria glabrata. To further facilitate progress toward control of schistosomiasis, a system for axenic in vitro culture of the parasite has now been developed. When culture media were preconditioned by Bge cells, sporocysts lived longer in vitro and produced more offspring. Because Bge-derived components could be protecting sporocysts from oxidative stress, axenic sporocysts were cultured at lowered O₂ levels. In an hypoxic environment, S. mansoni sporocysts grew well and produced daughter sporocysts continuously under axenic conditions and in a medium completely lacking host molecules. Sporocyst production occurs independently of host influence.

Three species of Planorbidae have been reported from Korea, e.g., Gyraulus convexiusculus, Hippeutis (Helicorbis) cantori, and Segmentina (Polypylis) hemisphaerula. Of these, only H. cantori was reported as the first and second intermediate host for Echinostoma cinetorchis, an important human intestinal parasite in Korea. Segmentina hemisphaerula has also been found to be an intermediate host. In field-collected planorbids, only S. hemisphaerula was found shedding echinostome cercariae and infected with metacercariae of E. cinetorchis, whereas no G. convexiusculus and H. cantori were found to be infected. In experiments with laboratory-bred snails, G. convexiusculus and S. hemisphaerula were susceptible to infection by miracidia of E. cinetorchis, but H. cantori could not be infected. Tadpoles of Rana nigromaculata and laboratory-bred snails of the 3 planorbid species were exposed to E. cinetorchis cercariae shed from field-collected S. hemisphaerula. All tadpoles, S. hemisphaerula, and G. convexiusculus became infected, but no H. cantori were infected. Metacercariae from tadpoles, S. hemisphaerula, and G. convexiusculus were fed to rats per os, and eggs of E. cinetorchis were detected in the rat feces 1 wk later. The rats were killed, and adult E. cinetorchis were recovered from the small intestines. This is the first report of G. convexiusculus as a potential first and second intermediate host and of S. hemisphaerula as a new first and second intermediate host for E. cinetorchis in Korea.

The dose-related infectivity of Sarcocystis neurona sporocysts and merozoites of 2 recent isolates of S. neurona was compared in gamma interferon knockout (KO) mice. Tenfold dilutions of sporocysts or merozoites were bioassayed in mice, cell culture, or both. All 8 mice, fed 1,000 sporocysts, developed neurological signs with demonstrable S. neurona in their tissues. Of 24 mice fed low numbers of sporocysts (100, 10, 1), 18 became ill by 4 wk postinoculation, and S. neurona was demonstrated in their brains; antibodies (S. neurona agglutination test) to S. neurona and S. neurona parasites were not found in tissues of the 6 mice that were fed sporocysts and survived for >39 days. One thousand culture-derived merozoites of these 2 isolates were pathogenic to all 8 mice inoculated subcutaneously (s.c.). Of the 24 mice inoculated s.c. with merozoites numbering 100, 10, or 1, only 3 mice had demonstrable S. neurona infection; antibodies to S. neurona were not found in the 21 mice that had no demonstrable organisms. As few as 10 merozoites were infective for cell cultures. These results demonstrate that at least 1,000 merozoites are needed to cause disease in KO mice. Sarcocystis neurona sporocysts were infective to mice by the s.c. route.

Adult nematode parasites were recovered from the heart blood of a deceased island fox (Urocyon littoralis) submitted for necropsy to determine the cause of death. Examination of the recovered nematodes supported the generic diagnosis of Angiocaulus, a parasite in Angiostrongylidae found in domestic and wild canids and mustelids. Specific diagnosis of the worms from the island fox as Angiocaulus gubernaculatus is based on the morphology of the dorsal ray in the copulatory bursa of the male worm and its comparison with published descriptions of Angiocaulus raillieti and A. gubernaculatus. Although A. gubernaculatus has been typically associated with mustelid hosts, its occurrence in the island fox indicates that the host distribution for the parasite may not be as restricted as previously believed.

Leishmania tropica, which is endemic in Turkey, is the causative agent of cutaneous leishmaniasis. Leishmania tropica promastigotes (2 × 10⁷) isolated from a patient with dermal leishmaniasis and reproduced in NNN medium were inoculated subcutaneously into the footpads of 10 Balb/c mice. Cutaneous leishmaniasis developed on the footpads of 4 mice approximately 45 days later. Leishmania tropica amastigotes were observed in smear slides and then cultivated in NNN medium. Balb/c mice are a suitable laboratory model for this isolate of L. tropica and thus a source of amastigotes for studies on the immunology, chemotherapy, and pathogenicity of cutaneous leishmaniasis.

To further study genetic heterogeneity of Mexican stocks of Trypanosoma cruzi, genomic Southern analyses from 54 Mexican isolates and 5 South American reference stocks were carried out. The membranes were hybridized with a homologous cDNA clone from the ribosomal protein S4 that identifies allelic bands from a single gene type locus. These allelic bands were sequentially numbered depending on their relative size. Mexican T. cruzi stocks were quite homogeneous: 31 cases (57%) showed a homozygous genotype 3/3, and 21 isolates (38%) exhibited the heterozygote genotype 2/3. Just 2 Mexican stocks (3%) showed a different genotype 2/5, but the potential parental homozygous 2/2 was never observed. Being that T. cruzi is a diploid organism, the apparent absence of the presumptive parental homozygous genotype 2/2 argues against sexual reproduction within the population, at least as a common event. Therefore, these data support a clonal population structure of T. cruzi in Mexico.

Ostertagia cf. kasakhstanica, the putative minor morphotype of the polymorphic ostertagiine O. bisonis, is reported in Bison bison from South Dakota. Descriptions of the synlophe and details of other diagnostic characters useful in identification and differentiation of this minor morphotype from other ostertagiines in wild and domestic ruminants from western North America are presented. Laterally, the cervical synlophe is consistent with Type Ib pattern among ostertagiines and is within the range of variation previously described for O. bisonis. Among recognized minor morphotypes of ostertagiines in wild and domestic ruminants from North America, O. kasakhstanica is distinct from O. kolchida, O. dikmansi, and O. arctica based on a tapering lateral synlophe. It is further distinguished from O. lyrata by the pattern of the cervical synlophe in the lateral field, structure of the genital cone and spicules, and dimensions of the esophageal valve. Although morphologically similar to T. davtiani/T. trifucata and T. boreoarcticus f. minor, it can be readily distinguished by the 2-1-2 bursal pattern, the structure of the spicules and genital cone, and the length of the esophageal valve. Ostertagia bisonis/O. cf. kasakhstanica circulates among pronghorn, mule deer, bison, and cattle in west-central North America.

In vitro excystation studies were carried out on the metacercariae cysts of Paragonimus heterotremus obtained from naturally infected crabs Potamon spp. The effects of elastase, trypsin, trypsin-dog bile, trypsin-bile salt, and dithiothreitol (DTT) were examined. The trypsin-dog bile medium stimulated maximum excystation. Of the media that contained 1 mM DTT, the optimum conditions for the excystation were shown to be pH 9, temperature of 39–40 C, and osmolarity of 250–350 mOsm. The DTT acceleration was antagonized by all of the following 6 protease inhibitors: leupeptin (0.5–4 μg/ml), l-trans-epoxysuccinyl leucylamido (4-guanidine) butane (1–8 μM), N-tosyl-l-phenylalanine chloromethyl ketone (0.1–0.4 mM), Nα-p-tosyl-l-lysine chloromethyl ketone (25–200 μg/ml), iodoacetic acid (0.5–4 mM), and phenylmethylsulfonyl fluoride (1–4 mM). These results suggest that a number of extrinsic and intrinsic factors may modulate excystation.

Feather mites (Astigmata) are distributed around the world, living on the feathers of birds, but their mechanisms for transmission among hosts are not fully understood. There is anecdotal evidence of feather mites attached to louseflies (Diptera: Hippoboscidae), suggesting that feather mites may use these flies as a mode of phoretic transmission among birds. Two bird–lousefly associations (alpine swift Apus melba–Crataerina melbae and feral pigeon Columba livia–Pseudolynchia canariensis) were inspected to test the hypothesis that feather mites use hippoboscid flies as major mode of transmission. Both bird species showed a high prevalence and abundance of feather mites and louseflies. However, no feather mites were found attached to the 405 louseflies inspected, although skin mites (Epidermoptidae and Cheyletiellidae) were found on louseflies collected from feral pigeons. This study suggests that feather mites do not use hippoboscid flies as a major mode of transmission among birds.

In Trypanosoma brucei, uptake of host transferrin is mediated by a heterodimeric, glycosylphosphatidylinositol-anchored receptor derived from the 2 expression site–associated genes 6 and 7 (ESAG6 and ESAG7). By using specific antibodies, it is shown here that T. evansi, a trypanosome species transmitted mechanically by biting flies, also expresses a transferrin receptor composed of ESAG6 and ESAG7. The cellular uptake of transferrin in T. evansi is completely inhibited with anti-T. brucei (ESAG6/7 heterodimer) antibodies. The demonstration of a functional ESAG6/7 transferrin receptor in T. evansi supports further its close relationship to T. brucei.

This study was undertaken to investigate the cryopreservation of Cryptosporidium parvum oocysts. Oocysts purified from mouse feces were suspended in distilled water, 10% glycerin, and 2.5% potassium dichromate. They were stored at −20 C and −80 C for 2, 7, and 30 days, respectively. In addition to the purified oocysts, the feces of C. parvum-infected mice were preserved under the same conditions described above. Purified and fecal oocysts were thawed at 4 C, and their viability was assessed by a nucleic acid stain, excystation test, tissue culture infectivity test, and infectivity to immunosuppressed adult mice. Oocysts purified from fecal material prior to cryopreservation lost most of their viability and all of their infectivity for tissue culture and mice. However, when oocysts were cryopreserved in feces, between 11.7 and 34.0% were judged to be viable and retained their infectivity for mice when stored at −20 C (but not −80 C) for 2, 7, and 30 days. Clearly, fecal material provides a cryoprotective environment for C. parvum oocysts stored at −20 C for at least 30 days.

The seroprevalence of antibodies to Toxoplasma gondii was investigated in trapped lynx (Lynx canadensis) and bobcats (Lynx rufus) from Québec, Canada. Forty-seven of 106 (44%) lynx and 4 of 10 (40%) bobcats had positive titers for T. gondii (≥25) by means of the modified agglutination test incorporating mercaptoethanol and formalin-fixed tachyzoites. Seroprevalence was significantly higher (P < 0.0001) in adult lynx than in juvenile lynx. The presence of antibodies to T. gondii in lynx and bobcats suggests that this organism is widespread in the wild and that exposure to wild felids and game animals from Québec may represent a potential source of infection for humans.

As part of the Puget Sound Ambient Monitoring Program of the Washington Department of Fish and Wildlife, serum samples from 380 harbor seals (Phoca vitulina) were tested for antibodies to Toxoplasma gondii in the modified agglutination test (MAT) incorporating formalin-fixed tachyzoites and mercaptoethanol. Antibodies to T. gondii were found in 29 of 380 (7.6%) seals with titers of 1:25 in 13, 1:50 in 14, and ≥1:500 in 2 seals. Results indicate natural exposure of these wild marine mammals to T. gondii oocysts.

Neospora caninum is an apicomplexan parasite that causes neonatal neuromuscular disease in dogs and abortions in cattle. Dogs are the only proven definitive host. Little is known about the prevalence of antibodies to this parasite in wildlife. Sera from 99 raccoons (Procyon lotor) were examined for agglutinating antibodies to N. caninum using the modified agglutination test employing formalin-fixed tachyzoites as antigen. Raccoons originated in Florida (n = 24, collected in 1996), New Jersey (n = 25, collected in 1993), Pennsylvania (n = 25, collected in 1999), and Massachusetts (n = 25, collected in 1993 and 1994). Ten (10%) had antibodies to N. caninum; 9 had titers of 1:50, and 1 (1%) had a titer of 1:100. The present study indicates that raccoons have minimal exposure to N. caninum. The sera were also tested for agglutinating antibodies to Toxoplasma gondii and 46 (46%) were positive; 16 had titers of 1:50, 8 had titers of 1:100, and 22 had titers of ≥1:500.

Taxonomic characters for identification of Turgida turgida Rudolphi, 1819, a parasitic nematode of the Virginia opossum Didelphis virginiana, were studied by scanning electron microscopy. The distinguishing feature of the cephalic end is the presence of numerous denticles, structures associated with the internal tooth, and 2 spongelike areas on the inner side of each pseudolabia. The posterior end of male T. turgida differs from that in other species of Physalopteridae by the number of caudal papillae (22), truncated postcloacal papilla, and patterns of ventrocaudal ornamentation.

The pathogenicity of Leucocytozoon caulleryi against specific-pathogen-free laying hens was investigated. Many large schizonts (second-generation schizonts) of L. caulleryi were seen in the ovary and oviducts of chickens. Edema and pressure atrophy of the adjacent tissues were associated with these schizonts. The eggshell-secreting portion of the uterus exhibited the most severe damage in the oviduct. This experiment reconfirms that L. caulleryi may stop egg production in laying hens, presumably as a result of damage to ovaries and oviducts.

Four monoxenous nematodes and 1 heteroxenous cestode were found in 4 species of introduced small mammals on isolated sub-Antarctic islands of the Indian Ocean. In the Kerguelen Archipelago, Syphacia obvelata, Passalurus ambiguus (Nematoda: Oxyuridae), and Rodentolepis straminea (Cestoda: Cyclophyllidae) were respectively found in the house mouse Mus musculus, the rabbit Oryctolagus cuniculus, and the black rat Rattus rattus. One accidental nematode, Trichostrongylus sp. (Nematoda: Trichostrongylidae), was also found in a black rat on Kerguelen. On Possession Island (Crozet Archipelago), R. straminea was present in the black rat. On Amsterdam Island, the brown rat R. norvegicus harbored 2 species, R. straminea and Nippostrongylus brasiliensis (Nematoda: Heligmonellidae). The small number of founder hosts and the depauperate terrestrial communities on these remote islands explain the low diversity in the helminth communities of these introduced mammals compared with continental populations.

Several potential mammalian reservoirs of sylvatic species of Trichinella were examined from Texas, New Mexico, and Arizona. During 1998–99, tongues were collected from a black bear (Ursus americanus) in Arizona; from 9 black bears, a coyote (Canis latrans), and a mountain lion (Felis concolor) in New Mexico; and from 154 coyotes, 32 raccoons (Procyon lotor), 13 opossums (Didelphis marsupialis), 4 ocelots (Leopardus pardalis), 3 bobcats (Lynx rufus), and 5 feral hogs (Sus scrofa) in southern Texas. Larvae of Trichinella murrelli were identified by a multiple-polymerase chain reaction analysis in 1 black bear (11.1%) from New Mexico and in 7 coyotes (4.5%) of Texas, whereas Trichinella spiralis larvae were detected in the black bear of Arizona. This is the first report of Trichinella infection in wildlife of New Mexico and Texas and extends the distribution of T. murrelli into the southwestern United States near the border of Mexico.

In the central Pacific, the nematodes Spirocamallanus istiblenni and S. monotaxis (Camallanidae) are common intestinal parasites of coral reef fishes. The 2 worm species are encountered in hosts in similar environments and are very similar morphologically. In recent published redescriptions, only 1 character was found to differ between the 2 species. Because that character had not previously been examined among worms of this family and because the original authors did not statistically test their results, the morphological data are here statistically reexamined for consistency with the differentiation between the 2 species. Both a discriminant function analysis and a hierarchical cluster analysis support the existing species designations based on morphological data. These analyses reaffirmed the conclusions of previous studies; i.e., the relative position of the male's second preanal papilla is the only diagnostic characteristic.

Onchocercosis, caused by Onchocerca flexuosa, was observed in red deer (Cervus elaphus) from Spain for the first time. Adult specimens of O. flexuosa were found in nodules in subcutaneous tissues in 42 of 125 (33%) red deer between October 1994 and September 1995; intensity of infection ± SD was 3.93 ± 5.26 nodules per infected host. A clear seasonal pattern in the distribution of nodules was observed, with higher values of prevalence and intensity in fall and winter in contrast to spring and summer. Significant differences were found among age groups in prevalence, but not in the mean intensity. No differences in infection were apparent between male and female adult red deer.

To quantify the occurrence of the dipteran parasite Batrachomyia strigapes, the ground frog assemblage associated with a large ephemeral wetland in Western Sydney was sampled between January 1999 and April 2000. Parasite infection was restricted to Uperoleia laevigata (5.1%, n = 1,492). Parasites were found under the parotoid glands, most frequently on the left side. After correction for frog size, infection was shown to significantly reduce frog weight. Despite reduced individual fitness, because the level of infection was low in the population, it is unlikely to have a major effect on the anuran population under current environmental conditions.

Six leeches (Placobdella ornata) were allowed to feed on a painted turtle (Chrysemys picta marginata) infected with Haemogregarina balli and subjected to a period of diapause before being allowed to feed on 2 laboratory-reared snapping turtles (Chelydra serpentina). Weekly examination of thin blood films revealed infections of the turtles at 130 days postfeeding. These observations provide support for broad host specificity of hemogregarine parasites of chelonians.

Adult Schistosoma mansoni parasites live in the bloodstream of their vertebrate hosts where they consume red blood cells. Hemoglobin, released from the ingested red blood cells, is degraded by a variety of parasite proteases, including Sm31 (cathepsin B) and Sm32 (schistosome legumain). In this study the localization pattern of the Sm31 and Sm32 enzymes in cercariae (the infectious life cycle stage) was examined. Antibodies generated against recombinant Sm31 and Sm32 recognize their respective proteins in Western blots of soluble parasite extracts. Highest levels are seen in adult female extracts, whereas the level of both proteins is below detection in cercarial extracts. However, in fixed, whole cercariae, both proteins are seen in the cecum and protonephridia. In the cecum, the staining pattern has a granular appearance, suggesting that the proteins are packaged in vesicles. In the protonephridial system, Sm31 and Sm32 are detected in all 8 flame cells in the cercarial body and in both flame cells in the cercarial tail. The distribution of the 2 proteins differs in the flame cells. Examination of immunostained cercariae using laser scanning confocal microscopy shows that whereas Sm31 is located in the tubule cell, Sm32 is found in both the tubule cell and its adjoining cap cell. These findings suggest that the proteins are involved in the proposed excretory and osmoregulatory roles of flame cells.

The population dynamics in the enteric connective tissues of eosinophils, mucosal mast cells (MMC), and in the mucosal epithelium of goblet cells were examined morphometrically in fixed ileal tissue of outbred Sprague Dawley rats during the first 32 days of infection with the tapeworm Hymenolepis diminuta. MMC and eosinophils were present in the lamina propria and submucosa; however, only eosinophils were also present in the muscularis externa. Eosinophilic infiltrate was first observed in the lamina propria at 15 days postinfection (dpi) and the numbers of eosinophils remained elevated through 32 dpi. Initial mucosal mastocytosis was detected on 6 dpi and MMC numbers continued to rise over the study period without reaching a plateau. Goblet cell hyperplasia occurred only at 32 dpi. In contrast to some intestinal nematode infections where these same 3 cell types are associated with the host's expulsion responses, H. diminuta is not lost by a rapid host response in the outbred Sprague Dawley rat strain used in these experiments. We suggest that either the induction of hyperplasia of these host effector cells in ileum tissue during H. diminuta infection is not capable of triggering parasite rejection mechanisms, or the function of the induced hyperplasia is necessary for some as yet unassociated physiological or tissue architecture change in the host's intestine.

Alterations in host phenotype induced by parasitic infection are often interpreted as either host or parasite adaptations, depending on which of the two appears to benefit. Mermithid nematodes typically castrate their insect hosts and, therefore, any change in host behavior has no further fitness consequences for the host; the adaptive value of the modified behavior must be assessed with respect to parasite fitness only. In a New Zealand stream, mermithid-infected nymphs of mayflies in the genus Deleatidium were disproportionately represented in drift samples compared with benthic samples, suggesting that infection by mermithids results in an increased tendency to drift. Drifting mayflies face a higher predation risk from trout, and the mermithid nematodes they harbor die if ingested by a fish. The change in mayfly behavior induced by mermithids thus appears to have negative fitness effects for the parasite, and one possible explanation for this phenomenon is that it is a nonadaptive, pathological side effect of infection.

Significant differences in the life histories of the human hookworms Ancylostoma duodenale and Necator americanus necessitate their differentiation for epidemiological studies and the design of control programs. Current methods of identification require time-consuming, labor-intensive techniques. A polymerase chain reaction (PCR)-based method that enables rapid species identification is described. The mitochondrial cytochrome oxidase I genes of both species were sequenced, and species-specific primer sets were designed. The primers were used in PCR to amplify 585-bp fragments of the cytochrome oxidase gene from individual hookworm eggs, larvae, and adults. The technique was also able to identify mixed infections containing equal amounts of eggs from each species. The technique is rapid, technically simple, and sensitive and will permit the accurate identification of human hookworms in epidemiological field studies.