During the past several weeks, I, like many of my predecessors, have read many speeches previously delivered at the annual meeting of the American Society of Parasitologists (ASP). These are wonderful, exciting papers, and I recommend them to you. The history of our Society and much of parasitological research and philosophy are contained in these speeches. An understanding of parasitology and specifically of the ASP in the 21st century necessitates an understanding of our roots. Just as proteomics, genomics, DNA, and Mendel are a continuum, so is our progression as a science and as a Society. This thought is not original with me. Several presidents have discussed this point in detail: Eloise Cram in 1956, “Stepping Stones in the History of the American Society of Parasitologists” (Cram, 1956); Martin Ulmer in 1978, “What's Past is Prologue” (Ulmer, 1978); Harry Hoogstraal in 1984, “ASP: Its Historic Role and Modern Opportunities” (Hoogstraal, 1985); and Mike Kemp in 1988, “Parasitology a Degenerate Discipline, Populated by Degenerate Scientists, Studying Degenerate Organisms?” (Kemp, 1989).

The spore of Myxosoma cerebralis is redescribed and compared with that of a close relative, M. cartilaginis. M. cerebralis is smaller, with greater variability in size and shape. It possesses a mucous envelope whereas M. cartilaginis does not. The surface architecture of M. cerebralis, as seen with the scanning electron microscope, differs in possessing a conspicuous circumsutural groove, prominent polar filament pores, and a mucous envelope.

Myxobolus cerebralis is the myxozoan parasite responsible for causing whirling disease in salmonid fish. Although the parasite was first described nearly 100 yr ago, it received relatively little attention until the discovery of its 2-host life cycle in the mid 1980s. This was the first, complete, myxozoan life cycle to be described, and it was greeted with some skepticism because it united 2 stages of M. cerebralis that were previously classified in 2 separate taxa. In the last decade, there has been a renewed interest in this parasite because whirling disease has been implicated in the decline of wild trout populations in several western states in the United States. Subsequent research efforts have dramatically increased the understanding of the biology of M. cerebralis and the numerous factors that affect the severity of whirling disease in salmonid hosts. These efforts also have provided a great deal of new information concerning interactions between M. cerebralis and its aquatic oligochaete host Tubifex tubifex. This review examines the current state of M. cerebralis in relation to 3 categories: the life cycle, the salmonid hosts, and the oligochaete host.

Numerous studies have demonstrated that parasites with complex life cycles frequently manipulate the phenotype of their hosts to increase their transmission rate. Little is known, however, concerning density-dependent processes within infrapopulations of manipulative parasites—whether parasites cooperate to manipulate the host, whether competition counteracts with these potential cooperative benefits, or both. Here we explored these ideas, focusing on the association between the manipulative trematode Microphallus papillorobustus and its second intermediate host, the gammarid Gammarus insensibilis. From the data collected in the field, we found no evidence that co-occurring M. papillorobustus individuals benefit from the presence of conspecifics; instead, individuals in larger infrapopulations suffered reduced size and fecundity. Thus, the net effect of increasing density suggests that competition rather than cooperation is the dominant force in infrapopulations of M. papillorobustus.

One-hundred twelve amphibians, including 51 blue-spotted salamanders, Ambystoma laterale, 30 eastern American toads, Bufo americanus americanus, and 31 northern leopard frogs, Rana pipiens, were collected during April–October 1996 from Waukesha County, Wisconsin and examined for helminth parasites. The helminth compound community of this amphibian assemblage consisted of at least 10 species: 9 in American toads, 8 in leopard frogs, and 3 in blue-spotted salamanders. American toads shared 7 species with leopard frogs, and 2 species occurred in all 3 host species. Although there was a high degree of helminth species overlap among these sympatric amphibians, statistically significant differences were found among host species and percent of indirect or direct–life cycle parasites of amphibian species individual component communities (χ² = 1,015, P < 0.001). American toads had a higher relative abundance of nematodes, 59%, than larval cestodes, 31%, and larval and adult trematodes, 10%, whereas leopard frogs had a higher relative abundance of larval cestodes, 71.3%, and larval and adult trematodes, 25.3%, than nematodes 3.4%. This is related to ecological differences in habitat and dietary preferences between these 2 anuran species. Helminth communities of blue-spotted salamanders were depauperate and were dominated by larval trematodes, 94%, and few nematodes, 6%. Low helminth species richness in this host species is related to this salamander's relatively small host body size, smaller gape size, lower vagility, and more fossorial habitat preference than the other 2 anuran species. Adult leopard frogs and toads had significantly higher mean helminth species richness than metamorphs, but there was no significant difference in mean helminth species richness among adult and metamorph blue-spotted salamanders. Considering adult helminths, the low species richness and low vagility of caudatans as compared with anurans suggest that local factors may be more important in structuring caudatan helminth communities of salamanders than of anuran hosts. Helminth species infecting salamanders may be more clumped in their geographic distribution as compared with anurans, and the role of other hosts and their parasites at the compound community level may be important in structuring helminth communities of salamanders.

This study examined whether ecoparasitic larval Unionicola foili exhibited a sex bias when infecting laboratory populations of the host insect Chironomus tentans and whether an association with male or female midges increased the likelihood of larval mites returning to the aquatic habitat. When laboratory populations of C. tentans were exposed to larval U. foili, there was a higher prevalence of mites among female hosts at emergence (17 of 30 males vs. 25 of 30 females infected by mites). However, there was no significant difference in the distribution or abundance of larvae among infected male (mean = 2.3 larvae per host) and female (mean = 2.6 larvae per host) midges. Larval mites parasitizing both male and female chironomids were more likely to return to water than could be expected by chance. Mite larvae infesting female C. tentans were more likely to return to water when female hosts deposited egg masses in water, suggesting that oviposition plays an important role in cueing larvae parasitizing female midges to detach. The mechanism responsible for increasing the likelihood that mites parasitizing male hosts return to water remains unclear. Future studies will address the possibility of parasite-mediated changes in host behavior.

Schistosomes and soil-transmitted helminths currently infect a third of the world's human population. An important feature of these parasitic infections is their focal distribution, which has significant implications for control. Only a few studies have been carried out at the microepidemiological scale, comparing infection levels among individuals or households within a single village. In this study, data are presented from a cross-sectional survey, examining all children attending a primary school in rural Côte d'Ivoire over several consecutive days for Schistosoma mansoni, soil-transmitted helminths, and intestinal protozoa. All houses in the main village were mapped, and school children were linked to these households for small-area spatial analyses. Comparison between the 260 school children who live within the main village and the 89 children who reside in nearby settlements revealed significant differences in the overall prevalence and intensity of infections with S. mansoni and hookworm, confirming the focal nature of these 2 parasites. On the other hand, S. mansoni and hookworm infections exhibited random spatial patterns within the main village. The validity of these results is discussed in the context of this epidemiological setting, drawing attention to the issue of scale. Our findings have direct implications for intervention because they call for a uniform, community-wide approach to control schistosomiasis and soil-transmitted helminthiasis. Implementation can be relatively straightforward, and the proposed control approach might be cost-effective and prove sustainable.

Botfly larvae (Cuterebra sp.) infesting spiny rats (Proechimys semispinosus) in 8 small islands in the Panama Canal were studied. Rats were live trapped monthly on each island from January 1991 through February 2000 and visually examined for the presence of bots. Overall, bot prevalence was 4.6% and differed statistically among island rat populations. Rats were simultaneously infested by as many as 4 bots. Overall bot intensity was 1.3 bots per infested rat and did not differ among islands. Mean bot density across all islands was 0.0111 and was greater during the dry seasons than during the rainy seasons, but it did not differ among islands. Bots were found during all the 12 calendar months, suggesting a multivoltine reproductive schedule. Although bot activity varied seasonally, there was little synchrony of bot activity among islands. Bot density was related negatively to rainfall but was not related to host density, suggesting that drier ambient conditions may promote reproduction by adult bot flies in this system.

Diplectanum monticellii n. sp. is described from the gills of Cynoscion leiarchus, a marine Sciaenidae, from Itacuruçá, Rio de Janeiro (type locality); Baía de Guaratuba, Paraná; and Pontal do Sul, Paraná in Brazil. The new species is characterized by the following features: bell-shaped male copulatory organ with a sleevelike base, accessory piece absent, vas deferens looping left intestinal cecum, vaginal aperture sinistroventral, and ventral anchor with elongate superficial and deep roots.

Argentinochondria patagonensis n. gen., n. sp. (Copepoda: Chondracanthidae) is described from adult specimens recovered from the smooth kingklip Genypterus brasiliensis Reagan, 1903, from San Jorge Gulf, Patagonia, Argentina. The new genus is placed in the Chondracanthinae. Females of the new genus differ from other members of this subfamily in having 2 pairs of modified legs, a bulbose head, a long neck region, and a trunk devoid of processes. Males have usual chondracanthid form, oral appendages as in female, except a longer paragnath, fewer teeth on maxilla, and 2 pairs of reduced legs.

Tur megistoproctus, a new species of Laelapinae, is described from the pelage of the echimyid rodent Proechimys dimidiatus from the Atlantic forests of Ilha Grande, south of Rio de Janeiro. Measurements and illustrations are included for females and males. Another laelapine mite species, Tur turki Fonseca, co-occurred with T. megistoproctus in our studies and was recorded from the same host individuals and localities. These 2 laelapine mite species appear to be exclusively associated with a complex of echimyid rodent species (subgenus Trinomys) in the Atlantic forests of southeastern Brazil.

This article focuses on the initiation pathway of mucin-type O-glycosylation in helminth parasites. The presence of the GalNAc-O-Ser/Thr structure, also known as Tn antigen, a truncated determinant related to aberrant glycosylation in mammal cells, and the activity of the UDP-GalNAc:polypeptide N-acetyl-galactosaminyltransferase (ppGaNTase), the enzyme responsible for its synthesis, were studied in species from major taxonomic groups. Tn reactivity was determined in extracts from Taenia hydatigena, Mesocestoides corti, Fasciola hepatica, Nippostrongylus brasiliensis, and Toxocara canis using the monoclonal antibody 83D4. The Tn determinant was revealed in all preparations, and multiple patterns of Tn-bearing glycoproteins were observed by immunoblotting. Additionally, the first evidence that helminth parasites express ppGaNTase activity was obtained. This enzyme was studied in extracts from Echinococcus granulosus, F. hepatica, and T. canis by measuring the incorporation of UDP-(³H)GalNAc to both deglycosylated ovine syalomucin (dOSM) and synthetic peptide sequences derived from tandem repeats of human mucins. Whereas significant levels of ppGaNTase activity were detected in all the extracts when dOSM was used as a multisite acceptor, it was only observed in F. hepatica and E. granulosus extracts when mucin-derived peptides were used, suggesting that T. canis ppGaNTase enzyme(s) may represent a member of the gene family with a more restricted specificity for worm O-glycosylation motifs. The widespread expression of Tn antigen, capable of evoking both humoral and cellular immunity, strongly suggests that simple mucin-type O-glycosylation does not constitute an aberrant phenomenon in helminth parasites.

Sarcocystis neurona, Neospora caninum, N. hughesi, and Toxoplasma gondii are 4 related coccidians considered to be associated with encephalomyelitis in horses. The source of infection for N. hughesi is unknown, whereas opossums, dogs, and cats are the definitive hosts for S. neurona, N. caninum, and T. gondii, respectively. Seroprevalence of these coccidians in 276 wild horses from central Wyoming outside the known range of the opossum (Didelphis virginiana) was determined. Antibodies to T. gondii were found only in 1 of 276 horses tested with the modified agglutination test using 1:25, 1:50, and 1:500 dilutions. Antibodies to N. caninum were found in 86 (31.1%) of the 276 horses tested with the Neospora agglutination test—the titers were 1:25 in 38 horses, 1:50 in 15, 1:100 in 9, 1:200 in 8, 1:400 in 4, 1:800 in 2, 1:1,600 in 2, 1:3,200 in 2, and 1:12,800 in 1. Antibodies to S. neurona were assessed with the serum immunoblot; of 276 horses tested, 18 had antibodies considered specific for S. neurona. Antibodies to S. neurona also were assessed with the S. neurona direct agglutination test (SAT). Thirty-nine of 265 horses tested had SAT antibodies—in titers of 1:50 in 26 horses and 1:100 in 13. The presence of S. neurona antibodies in horses in central Wyoming suggests that either there is cross-reactivity between S. neurona and some other infection or a definitive host other than opossum is the source of infection. In a retrospective study, S. neurona antibodies were not found by immunoblot in the sera of 243 horses from western Canada outside the range of D. virginiana.

To determine the extent to which splenic T cells were affected by Schistosoma mansoni infection, we investigated the ability of the T cells to produce interferon (IFN)-γ, as well as their chemotactic ability 7 wk PI. In this study, we report that splenic T cells from Balb/c mice with S. mansoni infections were capable of producing levels of IFN-γ comparable with splenic T cells from naive mice. However, the T cells exhibited altered chemotactic activity, as evidenced by an inability to respond to secondary lymphoid-tissue chemokine (SLC/CCL21). Although no difference in chemokine expression was found between the spleens of infected versus control mice, chemokine production was greater in the livers of infected versus control mice. Collectively, these data indicate that Balb/c mice with 7-wk S. mansoni infection possess splenic T cells with altered chemotactic activity and that the alterations may be a consequence of the granulomatous response in the liver.

Taenia solium has a complex life cycle. Its cysticercus can lodge in the brain, causing neurocysticercosis (NCC), and the adult tapeworm's survival in the intestine results in taeniasis. In this study, the in situ detection of previously described glycoprotein antigens used for serological diagnosis of NCC and the detection of other glycoconjugates was explored in cysticerci and the surrounding porcine tissue to understand their potential role in pathogenesis. Immunohistochemistry with an antiserum specific for glycoprotein antigens rich in N-linked carbohydrates and in situ histochemistry with a battery of lectins that have affinity to a variety of glycoconjugates were performed. The glycoconjugates rich in N-linked carbohydrates were detected in the vesicular fluid and tegument of the vesicular membrane and scolex, where the parasite has direct contact with the host tissues during cysticercosis and taeniasis, respectively. Additionally, as the inflammatory response progressed, the parasite's antigenic glycoproteins were also detected in the cytoplasm of inflammatory cells in the surrounding granuloma. In contrast, the spiral canal tegument, which will be exposed to intestinal enzymes in taeniasis, had N-acetyl-galactosamine–rich mucins. Thus, the differential saccharidic composition in T. solium metacestode structures may be important for the survival of the parasite in different host sites.

Trypanosoma cruzi, the causative agent of Chagas disease, is able to reproduce intracellularly in many host cell types while in the mammalian host. Although cellular immunity is known to be important in resistance to infection, the ability of immune cells to interfere with the completion of the intracellular growth cycle of T. cruzi has not been described. Using a tissue culture system to study the parasite growth cycle, we have found that spleen cells from infected mice are able to decrease the number of parasites released from infected fibroblasts. Spleen cells from mice infected for as few as 14 days and as long as 300 days display this inhibitory ability. Parasite egress from infected cells is inhibited by factor(s) released by immune cells during coculture with infected fibroblasts. Immune cell depletion studies indicate that the inhibitory activity requires the presence of both CD4 T cells and μ B cells. These results suggest a direct ability of immune cells to somehow interfere with the completion of the intracellular cycle, and this ability may play a role in control of this parasite.

The royal burial chamber of what is today the Collegiate-Basilica of St. Isidoro in León, Spain, built and remodeled between the 10th and 13th centuries and in the 20th century renamed the Kings' Pantheon, has 13 royal tombs that were opened in the presence of the Abbot-Prior of the Collegiate to enable a group of researchers to obtain all possible information from the royal remains. Several samples were sent to the Parasitology Unit of the Animal Pathology (Animal Health) Department at the Veterinary Faculty of León (Spain). In all the tombs, eggs and remains of nonparasitic mites were observed. In a piece of linen cloth from the bottom of 1 tomb, an Anoplocephala perfoliata egg was found. Furthermore, 4 mummified bodies were found. In 2 of these, those belonging to Infantes María and Fernando, Ascaris lumbricoides eggs were found and in the latter Trichuris trichiura eggs. We have not found in the literature reviewed any records of studies of this kind carried out in Spain.

The present study was designed to investigate the tolerance to low temperatures of 9 Trichinella isolates in rat muscle tissue. Nine groups of 24 rats were infected with encapsulated Trichinella spiralis, Trichinella nativa, Trichinella britovi, Trichinella murrelli, Trichinella T6, Trichinella nelsoni, and 3 nonencapsulated Trichinella pseudospiralis strains. Six rats from each of the groups were necropsied at 5, 10, 20, and 40 wk postinfection (wpi). Muscle tissues containing Trichinella larvae were exposed to temperatures of −18, −5, and 5 C for 1 or 4 wk, and afterward the reproductive capacity index (RCI) in mice was determined for the 9 individual Trichinella isolates. Only T. nativa muscle larvae were infective after freezing at a temperature of −18 C. At 5 wpi all encapsulated isolates, except for the tropical species T. nelsoni, remained infective after exposure to a temperature of −5 C for both 1 and 4 wk, whereas nonencapsulated T. pseudospiralis survived only 1 wk of exposure. All Trichinella spp. remained infective after exposure to a temperature of 5 C. Muscle larvae for all investigated species remained infective as long as they persisted in live rats during the experiment. Analysis of variance showed a significant effect of age on the temperature tolerance of encapsulated T. spiralis and nonencapsulated T. pseudospiralis. In addition, significant interaction between age of muscle larvae and length of exposure was found. In general Trichinella muscle larvae of medium age (10 and 20 wpi) tolerated freezing better than early and late stages of infection (5 and 40 wpi). This is the first study to demonstrate such a relationship between age of infection and temperature tolerance of Trichinella spp. muscle larvae.

The first morphological description is made of all stages of the life cycle of a Paragonimus species infecting humans in Colombia. Larval stages were obtained both in vitro and from field collections. Adult Paragonimus spp. are described. The aquatic snail Aroapyrgus sp. serves as an intermediate host of this species, both naturally and experimentally, yielding rediae and cercariae. Crabs (Hypolobocera bouvieri monticola and H. emberarum) were found to be the natural second intermediate hosts, and individuals of another crab species (Strengeria sp.) were also infected in the laboratory.

Exotic poeciliid fishes introduced into Hawaiian freshwaters are responsible for the introduction of several exotic parasites, of which the most important are Camallanus cotti and Bothriocephalus acheilognathi in terms of potential disease threat to native stream fishes. This roundworm and tapeworm are the most prevalent and abundant freshwater fish helminths in Hawaiian streams. This study examined the seasonal and yearly population structure of C. cotti and B. acheilognathi to determine if the tropical Hawaiian environment characterized by low climatic variability permits continuous opportunities for parasite transmission regardless of time of year. Camallanus cotti displayed seasonal differences in prevalence and mean abundance, whereas B. acheilognathi did not. Camallanus cotti prevalence and mean abundance were higher in the Hawaiian summer (47.7%, 0.79) than in winter (25.8%, 0.36). A seasonal relationship of C. cotti levels is likely explained by extensive rains associated with the Hawaiian winter season, which may act to decrease parasite transmission by flushing infected poeciliid hosts, intermediate copepod hosts, and possibly free-living infective worm stages downstream. Bothriocephalus acheilognathi displayed low prevalence and mean abundance in both summer (4.0%, 0.06) and winter (6.5%, 0.07), and it may be difficult to detect seasonal changes due to these low levels. Camallanus cotti prevalence and mean abundance remained relatively constant from the summer of 1995 to the summer of 1999, indicating that levels of this roundworm are stable in Waianu Stream. Whereas B. acheilognathi prevalence and mean abundance were low during the summer of 1995 and the summer of 1997, a dramatic peak in prevalence and mean abundance was observed in the summer of 1998 (41.2%, 1.06), with levels decreasing sharply in the summer of 1999 (4.4%, 0.07). It appears that B. acheilognathi also is present in stable populations at low levels, even though levels rose sharply during a single year.

The developmentally arrested life stage of the entomopathogenic nematode Steinernema feltiae is exposed to threats of survival, including desiccation. We adopted a comprehensive approach to the study of the molecular mechanisms of desiccation stress tolerance in S. feltiae IS-6. We identified, expressed sequence tags (ESTs) that are differentially expressed during desiccation stress in S. feltiae IS-6 infective juveniles using DNA subtractive hybridization. These ESTs included genes that are known to be stress related, genes that are homologous to hypothetical Caenorhabditis elegans proteins, and novel genes that may be involved in traits specific to S. feltiae. Expression pattern characterization revealed that all analyzed ESTs were induced during 8 and 24 hr of dehydration of S. feltiae IS-6. Our results unveiled some of the components of the genetic networks that are activated in S. feltiae IS-6 during dehydration and suggested a differing pattern of temporal regulation during nematode dehydration.

Ribosomal subunit protein 9 (rps9) is a nuclearly encoded protein that resides in the apicoplast organelle of Toxoplasma gondii. Two cis-acting regions within the rps9 transit domain (amino acids 38–49 and 79–86), when combined with the rps9 signal sequence, were necessary and sufficient for apicoplast targeting. To investigate proteins interacting with the rps9 leader sequence, parasites expressing rps9 leader constructs fused to a glutathione S-transferase (GST) reporter were prepared, and proteins associated with the leader constructs were purified from extracts by affinity chromatography. In addition to GST-containing peptides, proteins with apparent masses of 92, 90, 86, and 160 kDa were purified. Mass spectrometry data suggested that the 92- and 90-kDa polypeptides appear to be subtilisin-like proteins, whereas the 86-kDa polypeptide was identified as the molecular chaperone BiP of T. gondii.

The virulence of the oocysts of 7 Czech Toxoplasma gondii isolates was tested. The oocysts were obtained by experimental infection of cats with the tissue cysts of T. gondii isolates from dogs, cats, and rabbits. The cats shed the oocysts in feces, with prepatent periods of 3–5 days postinfection (PI); the patent period was 7–18 days. The number of oocysts shed varied between 0.94 million and 47 million, with 0.66 million–39 million oocysts found in the daily samples of excrement. The cats ceased oocyst production at 11–22 days PI. Sporulated oocysts were used to prepare infective doses of 1 to 10⁵ oocysts for oral infection of 10 mice. Deoxyribonucleic acid isolated from 4 T. gondii isolates was used in polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) for amplification of the ROP1 gene and restriction of the product of amplification by restriction endonuclease DdeI. On the basis of their biological characteristics, all 7 isolates belonged to the group of “avirulent” strains. In the PCR-RFLP tests, 2 isolates, K9 and K19, showed an “avirulent” strain pattern.

Spinitectus macrospinosus n. sp., a parasite of the channel catfish Ictalurus punctatus, from the Red and Assiniboine rivers in southern Manitoba, Canada, is characterized by spines reaching a maximum length of 21.8–26.8 μm (means for males and females, respectively) in the anterior esophageal region, arranged in 4 sectors in the anterior region of the body. Spines in this region of the body increase in size and decrease in number (from a maximum of 6 to 3 per sector). The combination of spine length, number, and arrangement and the position of the excretory pore (between spine rows 6 and 7) readily distinguishes it from all its North American congeners, including the species it is most similar to, S. carolini. It is different from S. gracilis and S. acipenseri in having a posteriorly directed vagina, an excretory pore between spine rows 6 and 7, a longer stoma, a right spicule with a terminal ventral barb, and a heart-shaped caudal mucron. Spinitectus macrospinosus is similar to other North American species of Spinitectus, e.g., S. carolini, S. micracanthus, S. mexicanus, S. osorioi, and S. humbertoi, in having the anterior rows of spines arranged in 4 sectors. It is also similar to S. micracanthus and S. carolini in possessing a relatively long stoma, a posteriorly directed vagina, a right spicule with a terminal ventral barb, and a heart-shaped caudal mucron. Furthermore, it is similar to S. carolini in possessing an “area rugosa” with 2 rows of precloacal cuticular cleats. In southern Manitoba, S. macrospinosus appears to mature only in the channel catfish. Reexamination of museum specimens revealed that the nematode also is found in I. furcatus in Kentucky Lake (Kentucky–Tennessee) and in I. lacustris in Lake Texoma (Oklahoma–Texas).

The traditional classification of the ligulid tapeworms into 2 genera, Ligula Bloch, 1782 and Digramma Cholodkovsky, 1914, remains controversial. Molecular data of sequences for the 5′ end of the nuclear 28S ribosomal ribonucleic acid (rRNA) gene, the mitochondrial cytochrome c oxidase subunit I (COI) gene, and the nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) gene, as well as the first internal transcribed spacer (ITS1) of the nuclear ribosomal deoxyribonucleic acid (DNA), were used to characterize Digramma and to investigate its relationship with Ligula. Digramma spp. exhibited identical sequences with Ligula intestinalis both in the 28S rRNA and the COI gene and differed from L. intestinalis by 0.7% in the ITS1 region and 7.4% in the ND1 gene, respectively. A high degree of genetic conservation within 28S ribosomal DNA, COI, ITS1, and even ND1 genes, was found in Ligula and Digramma. The low genetic divergence in the 4 genes between Ligula and Digramma indicates that Digramma is probably not an independent genus. Therefore, it is proposed that Ligula and Digramma should be considered as 2 species within the genus Ligula and the tapeworms of Digramma collected from diverse localities in China belong to the same species. The present study also suggests that ITS1 and ND1 sequences can act as useful genetic markers to distinguish Ligula and Digramma.

The entire life cycle of the Brazilian Echinostoma luisreyi n. sp., of the 37–collar spine E. revolutum group, has been observed under experimental conditions. The snail Physa marmorata serves as its first natural intermediate host. This species and the planorbid Biomplalaria glabrata act as experimental second intermediate hosts. The worm recovery rate was 51.3 and 0.6% for mice and hamsters, respectively, but the infection did not develop in the quail Coturnix coturnix. The natural vertebrate host is not known. The most important morphological character of the new species separating it from the other Echinostoma species studied is the oral corner spines that increase in size from the latero-oral to the ventro-oral regions. The ratio of the sizes of the smaller oral spines and the larger aboral ones was 1.7:1.0. Also, the excretory pore is radially wrinkled and dorsally subterminal. Images obtained by scanning electron microscopy confirmed the characters that differentiate the new species and the most closely related species.

Cryptosporidium galli Pavlasek, 1999, described from the feces of birds, is redescribed with additional molecular and biological data. Oocysts are ellipsoidal, are passed fully sporulated, lack sporocysts, and measure 8.25 × 6.3 μm (range 8.0–8.5 × 6.2–6.4 μm) with a length–width ratio of 1.30 (n = 50). Oocysts are structurally similar to those of Cryptosporidium baileyi described from chickens, but in addition to being considerably larger than oocysts of C. baileyi, these oocysts infect the proventriculus in a variety of birds and not the respiratory tract. Oocysts were successfully transmitted from chickens to chickens, and morphologically similar oocysts also were observed in a variety of exotic and wild birds (Order Passeriformes, Phasianidae, Fringillidae, and Icteridae). Molecular and phylogenetic analyses at the 18S rRNA, HSP70, and actin gene loci demonstrate that this species is genetically distinct from all known species and genotypes of Cryptosporidium and, thus, was named C. galli.

A new species of digenean found in the intestines of the steely-vented hummingbird Amazilia saucerrottei and the yellow-olive flycatcher Tolmomyias sulphurescens from the Area de Conservación Guanacaste, Guanacaste, Costa Rica, resembles members of the Prosthogonimidae in having a highly lobate ovary; an elongate cirrus sac containing the cirrus, pars prostatica, and internal seminal vesicle; no external seminal vesicle; 2 fields of extracecal vitelline follicles restricted to the area between the intestinal bifurcation and testes; and uterine loops occupying all available space in the hind body. The new species differs from all other members of the family in having genital pores opening laterally to the cecum, immediately anterior to the acetabular level, and markedly oblique rather than symmetrical testes. Consequently, we propose the new genus Whallwachsia for the species. Preliminary phylogenetic assessment suggests that the species is the sister group of all other prosthogonimids.

We propose Tinamutrema as a new genus for Brachylaima centrodes (Braun, 1901) Dollfus, 1935 and for T. canoae, as a new species inhabiting tinamus in the Area de Conservación Guanacaste, Costa Rica. Specimens from Costa Rica resemble B. centrodes in having an elongate body, pretesticular genital pore and terminal genitalia, intercecal uterine loops occupying all available space between the anterior testis and the intestinal bifurcation, an oral sucker width:pharynx width ratio of approximately 1:0.55, an oral sucker:ventral sucker width ratio of approximately 1:1, and vitelline follicles extending into the forebody closer to the pharynx than to the anterior margin of the ventral sucker and by living in the cloaca. They differ from B. centrodes in having vitelline follicles that do not extend as far anteriorly as those in B. centrodes, which extend anteriorly to the level of the anteriormost extent of the cecal “shoulders,” dense tegumental spination as opposed to sparse or no spination, relatively smaller cirrus with fewer spines, longer and more sinous pars prostatica, and forebody averaging 36% of total body length (TBL) as opposed to 42% TBL. Both species differ from other members of the Brachylaimidae in possessing a spinose cirrus and a cirrus sac containing both the cirrus and the pars prostatica. Preliminary phylogenetic assessment suggests that these traits are plesiomorphic, and thus the species are basal to the rest of the Brachylaimidae, whose diagnosis we emend accordingly.

We describe 2 new species of leucochloridiid-like brachylaimoid digeneans parasitizing a variety of birds in the Area de Conservación Guanacaste, Costa Rica, each of which we assign to a new genus. According to Pojmanska's (Pojmanska, T. 2002a. Superfamily Brachylaimoidea Joyeux & Foley, 1930. In Keys to the Trematoda, D. I. Gibson, A. Jones, and R. A. Bray [eds.]. CAB International and The Natural History Museum, London, U.K., p. 31–36.) key for the Brachylaimoidea, we are unable to place either species in any family. One species most closely resembles members of Leucochloridium by having well-developed suckers, lacking an esophagus, and having cecal shoulders, gonads at the posterior end, and the genital pore at posterior end of body but differs by having symmetrical testes, a posttesticular ovary, and a terminal genital pore; thus, we propose the genus Bakkeius for it. The second new genus resembles members of Michajlovia by having ventral genital pores but differs by having extracecal uterine loops in the forebody, a cirrus sac containing the pars prostatica and seminal vesicle, and gland cells surrounding the genital pore; thus, we propose Pojmanskia for it. These new genera must currently be treated as incertae sedis according to Pojmanska (op. cit.); however, we feel that future phylogenetic analyses will require emendation of the family diagnosis for Leucochloridiidae to include those taxa with terminal and ventral genital pores and with preovarian testes.

Specimens of a species of cyclocoelid digenean inhabiting Jacana spinosa from the Area de Conservación Guanacaste, Costa Rica, most closely resemble Haematotrephus facioi (Brenes and Arroyo, 1962) Yamaguti, 1971, in the same host from Aranjuez, Puntarenas Province, Costa Rica, in having confluent vitelline follicles posteriorly, diagnostic of Neohaematotrophus, and in pharynx length, ovary width, and cirrus sac on the sinistral side. The new species is also highly similar in appearance to H. gendrei Dubois, 1959, also inhabiting a jacanid (from West Africa), which has vitelline follicles confluent posteriorly, and extending anteriorly to the intestinal bifurcation and genital pore opening immediately posterior to the anterior margin of the pharynx. Like H. facioi, H. gendrei has a relatively much shorter and broader cirrus sac than does the new species. Examination of the holotype and paratype of H. facioi confirmed that the specimens from Guanacaste differ in having a longer body, a larger ovary and eggs, and smaller testes. They also have the ovary on the sinistral rather than the dextral side of the body, genital pore anterior to the pharynx rather than at or posterior to the level of the posterior margin of the pharynx, longer and thinner cirrus sac, and eggs without eyespotted miracidia. Half the eggs in both specimens of H. facioi have well-developed eyespotted miracidia, whereas the typical condition for cyclocoelids is for virtually all eggs to exhibit eyespotted miracidia. Both H. facioi and H. gendrei are transferred to Neohaematotrophus, along with the new species.

In this article, we describe the findings obtained using immunoblot analysis in the diagnosis of canine visceral leishmaniasis (VL) and its correlation with serological titer and clinical status. We found that all animals bearing amastigote forms recognized antigens with 29 and 32 kDa and that this pattern can be exploited for diagnostic and epidemiological purposes. The recognition of the 29- and 32-kDa antigens was verified even in seronegative dogs and preceded seroconversion in periods ranging from several months to 2 yr. We found a correlation between serological titer and parasite burden. Although no correlation between antigenic recognition pattern and clinical status was observed, immunoblot analysis proved to be a reliable test to detect antibodies against Leishmania sp. antigens in dogs from areas with endemic VL.

Inhalation of water contaminated with Naegleria fowleri may lead to a potentially fatal infection of the central nervous system known as primary amebic meningoencephalitis (PAM). Amphotericin B (AMB), an antifungal drug, is the only agent with established clinical efficacy in the treatment of PAM, though therapy with this drug is not always effective and has been associated with adverse effects on the kidneys and other organs. We investigated the activity of various therapeutic agents against N. fowleri in an attempt to identify other useful agents for treating PAM. Several of these agents exhibited in vitro activity against the Lee (M67) strain of N. fowleri. The minimum inhibitory concentrations of these agents were 0.1 μg/ml (ketoconazole), 1 μg/ml (liposomal AMB), and 10 μg/ml (minocycline, quinupristin–dalfopristin, and trifluoperazine). Other agents had a minimum inhibitory concentration >10 μg/ml (linezolid) or >100 μg/ml (rifampin). In a mouse model of PAM, none of the untreated control mice survived, whereas the survival of treated animals was 50% (quinupristin–dalfopristin), 30% (ketoconazole and liposomal AMB), 20% (trifluoperazine), and 10% (linezolid and minocycline). Further studies are needed to ascertain whether these agents have synergistic activity with AMB in vitro and in vivo.

Current methods for the serodiagnosis of sheep fascioliasis show suboptimal sensitivity, specificity, or both. With the aim of developing an improved method, we fractionated native Fasciola hepatica excretory–secretory antigens (ESAs) by size-exclusion FPLC (fast protein liquid chromatography) on a Superdex™ 75 HR 10/30 column and then tested the serodiagnostic value of the antigens contained in each one of the 4 peaks obtained (peaks I–IV). Serodiagnostic value was assessed using sera from sheep naturally infected with F. hepatica (group A); sera from the individuals of a fluke-free herd (most of which also had other intestinal nematodes, lung nematodes, Moniezia spp., and/or Cysticercus tenuicollis) sera from a fluke-free herd (group B); sera from lambs experimentally infected with 10–40 F. hepatica metacercariae (group C); and sera from uninfected control lambs (group D). Enzyme-linked immunosorbent assay (ELISA) with peak I or II as target antigens (and to a lesser extent with peak III as target) showed reactivity with negative sera, so that it was not possible to establish cutoff values discriminating infected and uninfected animals. In contrast, when peak IV was used as target, a low cutoff value of 0.235 optical density units (mean 4 SD) discriminated infected and uninfected animals, with 100% sensitivity and 100% specificity. ELISA with peak IV as a target identified infected animals (even animals that had received only 10 metacercariae) within 3–5 wk of infection and subsequently throughout the rest of the 14-wk monitoring period. In Western blotting analysis, again only the antigens contained in peak IV (range 7–40 kDa, under reducing conditions) were specific for diagnosis of infected animals. These results indicate that molecular sieving of F. hepatica ESAs by this procedure is a fast, simple, reproducible way of obtaining antigens useful for serodiagnosis of sheep fascioliasis.

Capybaras (Hidrochoeris hidrochoeris) are 1 of the largest rodents used for meat in South and Central America. Prevalence of anti–Toxoplasma gondii antibodies in 149 feral H. hidrochoeris from the state of São Paulo, Brazil, was evaluated using the indirect immunofluorescent antibody test (IFAT) and the modified agglutination test (MAT). Using IFAT, antibodies (>1:16) were found in 104 (69.8%) and with the MAT, antibodies (>1:25) were found in 63 (42.3%) capybaras. This is the first report of prevalence of T. gondii antibodies in this host.

Most isolates of Toxoplasma gondii can be grouped into 3 genetic lineages. In the present study, 67 isolates of T. gondii were obtained by bioassay in mice inoculated with brains and hearts of 96 asymptomatic chickens from an area highly endemic to human infection in Rio de Janeiro, Brazil. Of the 48 isolates genotyped using the SAG₂ locus, 34 (70%) were of type I and 13 (27%) were of type III. No isolate of type II was recovered. Isolates from 1 chicken contained a type I and type III mixed infection, indicating natural multiparasite infection in the same animal. Cats fed mice infected with 11 type I strains shed 19–535 million oocysts in their feces, indicating that type I isolates can circulate in the environment.

Laboratory dogs were vaccinated intramuscularly with a recombinant fusion protein (expressed and isolated from Escherichia coli) formulated with the Glaxo SmithKline Adjuvant System 02 (AS02). The fusion protein encoded Ac-MTP-1, a developmentally regulated astacinlike metalloprotease secreted by host-stimulated Ancylostoma caninum third-stage larvae (L3). Control dogs were injected intramuscularly with an equivalent amount of AS02 adjuvant alone. The vaccinated and control dogs were then challenged by s.c. injection of 500 L3 of the canine hookworm A. caninum. The vaccinated dogs developed prechallenge immunoglobulin G2 (IgG2) antibody responses specific to anti–Ac-MTP-1-fusion protein with titers ranging between 1:40,000 and 1:364,000, whereas they developed antigen-specific immunoglobulin E antibody responses with titers ranging between 1:500 and 1:1,500. By immunoblotting, canine sera obtained from the vaccinated dogs recognized a protein of the estimated apparent molecular weight of Ac-MTP-1 in activated L3 secretory products. Spearman rank order correlations between the canine intestinal adult hookworm burden and quantitative egg counts at necropsy and anti-Ac-MTP-1 IgG2 antibody titers revealed a statistically significant inverse association (r = −0.89; P = 0.04), suggesting that this molecule offers promise as a recombinant vaccine.

The present study was conducted to determine the effect of multiple low doses of methylmercury (MeHg) on the course of a chronic Toxoplasma gondii infection. Four groups of 6-wk-old female CBA/J mice either were fed 25 T. gondii tissue cysts of the ME-49 strain or were vehicle control. Six weeks later, half of each group was orally gavaged with 8-mg/kg body weight doses of MeHg on days 0, 2, 4, 7, 10, and 13, totaling 4 experimental groups. Mice were killed on day 17 or 18 after MeHg exposure. Flow cytometric analysis of lymphocyte subpopulations in the thymus demonstrated a significant increase in the percentage of CD4⁻CD8 T-cells in mice exposed to MeHg with a concurrent T. gondii infection. Groups of mice exposed to MeHg showed a decrease in total thymic cellularity and cellularity of all T-cell subpopulations when compared with control mice, but viability of these cells was unaffected. Splenic cell viability was decreased in mice exposed to MeHg, but alterations in T-cell subpopulations were not noted. These data indicate that multiple low doses of MeHg may not exacerbate chronic toxoplasmosis, but MeHg-induced effects on the immune system were evident.

A Pacific harbor seal (Phoca vitulina richardsii) was found on the central California coast with neurologic signs and labored breathing, which were unresponsive to treatment. Necropsy revealed a nonsuppurative necrotizing meningoencephalitis, a multilocular thymic cyst, and nonsuppurative cystitis and renal pyelitis. Microscopic examination revealed protozoans in the brain, thymic cyst, and bladder mucosa. Ultrastructurally, the protozoal tachyzoites were different from those of Neospora caninum, Toxoplasma gondii, and Sarcocystis neurona; the rhoptries were small and had electron-dense contents, and the organism divided by endodyogeny. Specific antibodies were not detected in serum using agglutination (N. caninum, T. gondii) and immunoblot assays (S. neurona). Immunohistochemistry for these organisms was negative. Polymerase chain reaction on brain tissue using specific primers did not amplify T. gondii deoxyribonucleic acid. The meningoencephalitis in this seal thus appears to have been caused by a novel protozoan.

The whelk Cominella glandiformis is an important predator–scavenger of New Zealand intertidal ecosystems; a few whelks can quickly eat all the soft tissues of recently dead crabs. In this study, we demonstrate that whelks can also ingest and act as paratenic hosts for at least 4 helminth species that use crabs as intermediate hosts: metacercariae of the trematode Maritrema sp. and of another unidentified trematode, larval acuariid nematodes, and cystacanths of the acanthocephalans Profilicollis spp. Large whelks ingest disproportionately more helminth larvae than small whelks, but the survival of parasites during their short stay in the whelks is not affected by whelk size. The majority of metacercariae and nematodes are passed out in whelk feces within 3 days of ingestion, whereas the few cystacanths found did not leave whelks until after that time; no parasite was left in whelks 5 days postingestion. Survival of all 4 helminth species was generally very high, though it decreased day by day in 2 species. Given that the avian definitive hosts of all 4 helminths also eat whelks, our results indicate that alternative transmission pathways exist and that parasites can take routes through food webs that are too often ignored.

This is the first report of blue autofluorescence as a useful characteristic in the microscopic detection of Toxoplasma gondii, Hammondia hammondi, Hammondia heydorni, Neospora caninum, Besnoitia darlingi, and Sarcocystis neurona oocysts or sporocysts. This autofluorescence is of sufficient intensity and duration to allow identification of these oocysts from complex microscopic sample backgrounds. As with the autofluorescence of related coccidia, the oocysts glow pale blue when illuminated with an ultraviolet (UV) light source and viewed with the correct UV excitation and emission filter set.

The nematode parasites Wuchereria bancrofti, Brugia malayi, and B. timori cause a disease in humans known as lymphatic filariasis, which afflicts approximately 120 million people worldwide. The parasites enter the human host from the mosquito either as L3 or as infective larvae and subsequently differentiate through 2 molts. In this article, we show that B. malayi depends on an exogenous source of vitamin C to complete the L3 to L4 molt, a critical morphogenic step in its life cycle. Brugia malayi apparently belongs to a small group of living organisms that depend on an exogenous source of vitamin C. This group includes only primates (including man) and guinea pigs among mammals.

Antibodies to Neospora caninum and Toxoplasma gondii were assayed in sera of 396 opossums (Didelphis marsupialis) from the city of São Paulo, Brazil. Antibodies to N. caninum were assayed using the indirect immunofluorescent antibody test (IFAT). Antibodies (IFAT, ∼1:25) to N. caninum were found in 84 opossums (D. marsupialis) in titers of 1:25 in 46, 1:50 in 20, 1:100 in 17, and 1:400 in 1. Antibodies to T. gondii were assayed with the modified agglutination test (MAT) and the IFAT. Antibodies to T. gondii (MAT, ∼1:25) were found in 82 (20.4%) of the 396 opossums, in titers of 1:25 in 24, 1:50 in 26, 1:100 in 18, 1:200 in 13, and 1:800 in 1. The IFAT antibodies to T. gondii were found in 148 of 396 opossums, in titers of 1:16 in 41, 1:32 in 23, 1:64 in 13, 1:128 in 6, 1:256 in 20, 1:512 in 17, 1:1,024 in 10, 1:2,048 in 10, 1:4,096 in 7, and 1:8,192 in 1. This is the first report of N. caninum and T. gondii infections in D. marsupialis.