As part of an investigation of the biochemical interactions between the salmon louse Lepeophtheirus salmonis and Atlantic salmon Salmo salar, we characterized protease activity in the skin mucus of noninfected Atlantic salmon and Atlantic salmon infected with L. salmonis and in an L. salmonis whole-body homogenate. Zymography revealed that mucus from infected salmon contained a series of low-molecular-mass (17–22 kDa) serine proteases that were not present in the mucus of noninfected salmon. Based on molecular mass, inhibition studies, and affinity chromatography, the series of proteases was identified as being trypsin-like. Similar proteases were observed in the L. salmonis homogenate and in mucus from noninfected Atlantic salmon following a 1-hr incubation with live L. salmonis. An antibody raised against Atlantic salmon trypsin failed to recognize any proteases in the mucus of noninfected salmon or infected salmon or in the L. salmonis homogenate. Collectively, these findings suggest that the trypsin-like proteases present in the mucus of infected Atlantic salmon were produced by L. salmonis, possibly to aid in feeding and evasion of host immune responses.

In Ascaris suum, muscle glycogen is synthesized during host feeding intervals and degraded during nonfeeding intervals. Glycogen accumulation is up to 12-fold greater than that observed in mammalian muscle. Previous studies have established that many aspects of the parasite glycogen metabolism are comparable with the host, but a novel form of glycogen synthase designated GSII also occurs in the parasite. In this report glycogenin has been identified as the core protein in both mature glycogen and the GSII complex. Digestion of GSII complex glycogen generates discreet intermediates that may correspond to a proglycogen pool, whereas digestion of mature glycogen does not generate these intermediates. Because both GSII complex glycogen and mature glycogen serve as GSII substrates, the GSII complex likely represents an intermediate between glycogenin and mature glycogen. The regulation of glycogenin synthesis or the regulation of GSII activity that converts glycogenin to proglycogen, or both, may account for high levels of polysaccharide accumulation that are essential for A. suum survival.

Differentiation of Trypanosoma cruzi epimastigotes to metacyclic trypomastigotes occurs in the insect rectum, after adhesion of the epimastigotes to the intestinal wall. We investigated the effect of the nutritional stress on the metacyclogenesis process in vitro by incubating epimastigotes in the chemically defined TAU3AAG medium supplemented with different nutrients. Addition of fetal bovine serum induced epimastigote growth but inhibited metacyclogenesis. In this medium, few parasites attached to the substrate. Ultrastructural analysis demonstrated reservosomes at the posterior end of the epimastigotes. Incubation of the cells in TAU3AAG medium containing gold-labeled transferrin resulted in high endocytosis of the marker by both adhered and free-swimming epimastigotes. No intracellular gold particles could be detected in trypomastigotes. Addition of transferrin–gold complexes to adhered epimastigotes cultivated for 4 days in TAU3AAG medium resulted in decrease of both metacyclogenesis and adhesion to the substrate, as compared with parasites maintained in transferrin-free medium. Adhesion to the substrate is triggered by nutritional stress, and proteins accumulated in reservosomes are used as energy source during the differentiation. A close relationship exists among nutritional stress, endocytosis of nutrients, adhesion to the substrate, and cell differentiation in T. cruzi epimastigotes.

Swingleus ancistrus n. sp. (Monogenea: Gyrodactylidae) is described from the skin and fins of Fundulus heteroclitus from Canary Creek Marsh, Lewes, Delaware. Subsequent records from New Jersey, Maryland, Virginia, and North Carolina are reported. Swingleus ancistrus is differentiated from S. polyclithroides, the only other species of the genus, primarily by its size, haptoral morphology, host, and locality. Swingleus ancistrus is larger in almost every dimension and exhibits a distinct notch on the anterior border of the opisthaptor, and several features of the haptoral anatomy and peduncular bar are unique. The prevalence of infection in hosts collected from the type locality in August was 100%. The maximum number of S. ancistrus recovered from a single host was 134. The average intensity of infection was 24 S. ancistrus per host from the type locality.

The potential bactericidal activity of the alternative complement pathway of mammalian and reptilian sera to Borrelia burgdorferi sensu stricto (s.s.) was evaluated in vitro. Complement-mediated killing was observed when cultured spirochetes were inoculated into sera from the western fence lizard (Sceloporus occidentalis) and from the southern alligator lizard (Elgaria multicarinata), but not when they were inoculated into serum from either the deer mouse (Peromyscus maniculatus) or from humans. Spirochetes were still alive after 4 hr in lizard serum that had been preheated at 56 C for 30 min to inactivate complement. Furthermore, when lizard serum was chelated with 10 mM ethylenediaminetetraacetic acid to block all complement activation, borreliacidal activity was arrested. When lizard serum was chelated with 10 mM ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid plus 4 mM MgCl₂ to block only classical complement pathway activation, >85% of spirochetes were immobilized within 1 hr. Differences in B. burgdorferi s.s. mortality were not observed when chelators with or without MgCl₂ were added to serum from either deer mice or humans. Proteins comprising the alternative complement pathway are responsible for the borreliacidal activity observed in the blood of S. occidentalis and E. multicarinata.

A new species of trichosomoidid nematodes, Huffmanela mexicana n. sp., is established on the basis of its egg morphology and biological characters. The dark-shelled, embryonated eggs of this histozoic parasite occur in masses in the inner layer of the swimbladder wall of the bullseye puffer fish Sphoeroides annulatus Jenyns, 1843 (Tetraodontidae, Tetraodontiformes) off the Pacific coast of Mexico (Sinaloa State) and are characterized mainly by their shape and size (63–69 × 30–33 µm), an aspinose surface, relatively small polar plugs, and their thick egg wall (4–5 µm). A key to Huffmanela species based on egg morphology is presented.

Various species of cervid deer are the preferred hosts for adult, black-legged ticks (Ixodes scapularis and Ixodes pacificus) in the United States. Although frequently exposed to the agent of Lyme disease (Borrelia burgdorferi), these animals, for the most part, are incompetent as transmission reservoirs. We examined the borreliacidal activity of normal and B. burgdorferi-immune sera from sika deer (Cervus nippon) maintained in a laboratory setting and compared it to that of similar sera from reservoir-competent mice and rabbits. All normal deer sera (NDS) tested killed >90% of B. burgdorferi cells. In contrast, normal mouse and rabbit sera killed ≤22% of the Borrelia. Anti-B. burgdorferi antibodies could not be detected in any normal sera by indirect fluorescent antibody assay (IFA). Sera collected from deer 6 wk after exposure to B. burgdorferi by tick feeding exhibited IFA titers of 1:256, whereas sera from mice and rabbits similarly exposed had titers of >1:1,024. Heat treatment (56 C, 30 min) of NDS reduced borreliacidal activity, with <20% of the B. burgdorferi cells killed, suggesting complement-mediated killing. The chelators EGTA and EDTA were used to block the classical or both the classical and alternative complement pathways, respectively. Addition of 10 mM EGTA to NDS had a negligible effect on borreliacidal activity, with >90% of the cells killed. Addition of 10 mM EDTA reduced the killing to ∼30%, whereas the addition of Mg² (10 mM) restored borreliacidal activity to NDS. The addition of zymosan A, an activator of the alternative pathway, increased the survival of B. burgdorferi cells to ∼80% in NDS. These data suggest that the alternative complement activation pathway plays a major role in the borreliacidal activity of NDS. Additionally, 10 mM EGTA had almost no effect on the killing activity of B. burgdorferi-exposed deer sera, suggesting that the classical pathway is not involved in Borrelia killing, even in sera from B. burgdorferi-exposed deer.

The morphology and patterns of distribution of the argentophilic structures of miracidia and cercariae of Philophthalmus distomatosa n. comb. are described. The epidermal plate arrangement of the vast majority (94.2%) of miracidia studied conformed to the formula 6:8:4:2 = 20. The rest (5.8%) displayed the following patterns of arrangement of epidermal plates: 6:7:4:2 = 19; 6:6:4:2 = 18; and 6:4:4:2 = 16. Twenty papillalike structures were observed on the terebratorium. They were arranged along 1 axis, in 3 groups. Most commonly, 16 papillae were present on the body, located between epidermal plates of the first and second rows. The eye spots were located dorsally, near interepidermal space S₁. The numbers, arrangements, and locations of the excretory pores varied. Patterns of distribution of the tegumentary papillae of P. distomatosa n. comb. cercariae are given, including those of the cephalic regions, periacetabular region, and tail. Argentophilic structures of the mentioned larval stages and of other parameters clearly distinguishing between the presently described species and those of P. lucipetus from Israel are summarized. On the basis of comparison of the presently described larval stages and those of other Melanoides tuberculata-associated larval stages of Philophthalmus in Israel and Jordan, the possibility exists that a third species occurs in Israel.

This study was undertaken in order to characterize a Cryptosporidium muris-like parasite isolated from cattle in Hungary and to compare this strain with other Cryptosporidium species. To date, the large-type oocysts isolated from cattle were considered as C. muris described from several mammals. The size, form, and structure of the oocysts of the Hungarian strain were identical with those described by others from cattle. An apparent difference between the morphometric data of C. muris-like parasites isolated from cattle or other mammals was noted, which is similar in magnitude to the differences between Cryptosporidium meleagridis and Cryptosporidium felis or between Cryptosporidium serpentis and Cryptosporidium baileyi. The cross-transmission experiments confirmed the findings of others, as C. muris-like oocysts isolated from cattle fail to infect other mammals. The sequence of the variable region of small subunit (SSU) rRNA gene of the strain was 100% identical with that of the U.S. Cryptosporidium andersoni and C. andersoni-like isolates from cattle. The difference between the SSU rRNA sequence of bovine strains and C. muris is similar in magnitude to the differences between C. meleagridis and Cryptosporidium parvum anthroponotic genotype or between Cryptosporidium wrairi and C. parvum zoonotic genotype. Our findings confirm that the Cryptosporidium species responsible for abomasal cryptosporidiosis and economic losses in the cattle industry should be considered a distinct species, C. andersoni Lindsay, Upton, Owens, Morgan, Mead, and Blagburn, 2000.

Intraspecific heterogeneity was demonstrated in the mini-exon gene localization from Leishmania (Viannia) panamensis and L. (Viannia) guyanensis. Different karyotypes were detected in human isolates circulating in endemic areas of Colombia. The presence of mini-exon gene sequences on chromosomes of different sizes, ranging from 370 to 800 kb in L. (V.) panamensis and from 500 to 800 kb in L. (V.) guyanensis, was observed and was neither strain nor species specific. In some cases, hybridization with 2 chromosomes in the same strain was observed. The variability of chromosomal localization of mini-exon gene sequences of these 2 species highlights the genetic variability of the Viannia subgenus and the potential utility of the mini-exon gene as a molecular epidemiologic marker.

Unionicola formosa is a symbiotic water mite that passes most of its life cycle in the mantle cavity of freshwater mussels. Although mites of this genus are often referred to as parasitic, little is known about their nutritional biology. A few species reportedly pierce the gill of a host mussel and ingest tissue or hemolymph. The present study was undertaken to identify possible sources of nutrition for U. formosa. To determine if mites ingested particulate matter in the mucous strand produced by a mussel during feeding, mussels with resident mites were exposed to a suspension of fluorescent microspheres. There was no evidence that U. formosa ingested the beads. Histochemical staining did, however, indicate a mucous material present in the midgut of the mites. Sodium dodecyl sulfate–polyacrylamide gel electrophoretic assays revealed a high molecular weight component, consistent with a mucopolysaccharide, present both in the mussel gill and the mites. Results from western blots and an immunoaffinity binding assay with antibodies against mussel gill tissue and hemolymph also indicated that mites ingested host tissue. Whereas U. formosa probably does not ingest particulate material acquired by its host's suspension feeding, it is apparent that this mite utilizes host mucus, gill tissue, or hemolymph for at least part of its nutrition.

The Drosophila–parasitic wasp (parasitoid) associations involve integrating adaptations of considerable complexity. This review focuses on some of the factors that influence these interactions including host immunity, nutrition and hormonal changes, and parasitoid virulence and mechanisms of immune suppression.

The growth and emigration of Hystherothylacium aduncum in laboratory-reared herring larvae Clupea harengus was studied. Experimental infections of 36-day-old herring larvae resulted in 126 hosts infected with 306 H. aduncum larvae. Regression analyses showed a significant worm emigration from the rectum to the head of the fish, accompanied by an increase in worm body length. The emigration was independent of worm intensity, which suggests an ontogenetic process. Some worms departed from this pattern by moving posteriorly or by penetrating into the muscle, and in 5 cases, the larvae were observed to leave living fish. This individual variation has not been observed in previous studies and might be explained by host signals related to condition or development stage. Indirect evidence suggested parasite-induced mortality in the tanks due to the emigrations because only 4 of the 126 infected fish survived 8 days postinfection; the emigration of H. aduncum affected vital organs, such as the heart and brain, and the larvae penetrating or leaving the host's tissues can cause extensive damage to the delicate herring larvae.

Sarcocystis neurona is the most important cause of a neurologic disease in horses, equine protozoal myeloencephalitis (EPM). The complete life cycle of S. neurona, including the description of sarcocysts and intermediate hosts, has not been completed until now. Opossums (Didelphis spp.) are definitive hosts, and horses and other mammals are aberrant hosts. In the present study, laboratory-raised domestic cats (Felis domesticus) were fed sporocysts from the intestine of a naturally infected opossum (Didelphis virginiana). Microscopic sarcocysts, with a maximum size of 700 × 50 µm, developed in the muscles of the cats. The DNA of bradyzoites released from sarcocysts was confirmed as S. neurona. Laboratory-raised opossums (D. virginiana) fed cat muscles containing the sarcocysts shed sporocysts in their feces. The sporocysts were ∼10–12 × 6.5–8.0 µm in size. Gamma interferon knockout mice fed sporocysts from experimentally infected opossums developed clinical sarcocystosis, and S. neurona was identified in their tissues using S. neurona-specific polyclonal rabbit serum. Two seronegative ponies fed sporocysts from an experimentally-infected opossum developed S. neurona-specific antibodies within 14 days.

The ribosomal RNA (rRNA) gene promoter was used to construct plasmid vectors that simultaneously express multiple exogenous genes in Trypanosoma cruzi. Vector pBSPANEO expresses neomycin phosphotransferase, and pPAGFPAN expresses both green fluorescent protein and neomycin phosphotransferase from a single promoter. Both vectors require the presence of the rRNA promoter for stable transfection; epimastigotes transfected with pPAGFPAN strongly fluoresced due to green fluorescent protein expression. Intact plasmids were rescued from the T. cruzi-transfected population after >8 mo of culture, indicating stable replication of these vectors. Vectors were integrated into the rRNA locus by homologous recombination and into other loci, presumably by illegitimate recombination. Parasites bearing tandem concatamers of plasmids were also found among the transfectants. Transfectants expressing green fluorescent protein showed a bright green fluorescence distributed throughout the cell. Fluorescence was also detected in amastigotes after infection of mammalian cells with transfected parasites, indicating that the rRNA promoter can drive efficient expression of these reporter genes in multiple life-cycle stages of the parasite. Expression of the heterologous genes was detected after passage in mice or in the insect vector. These vectors will be useful for the genetic dissection of T. cruzi biology and pathogenesis.

The kinetoplast DNA (kDNA) minicircle molecules of 14 Brazilian stocks of Trypanosoma evansi were studied by morphological approaches (Giemsa and 4′-6′-diamidino-2-phenylindole staining and transmission electron microscopy) and molecular approaches (probing with an oligonucleotide complementary to the minicircle origin of replication and polymerase chain reaction amplification of a minicircle sequence). All methods indicated the absence of both a typical kinetoplast and kDNA minicircles, even in a very small number of parasites of a single stock or in small numbers of copies of molecules per cell. We did not detect any altered kDNA molecules. There were no kDNA molecules in either old or new stocks of T. evansi maintained by successive passages in mice. Similarly, no kDNA minicircles were detected in trypanosomes in blood smears from naturally infected domestic and wild animals. Thus, the total absence of kDNA in Brazilian stocks of T. evansi from both domestic and wild mammals is probably the natural state of Brazilian T. evansi.

Clinical coccidiosis was diagnosed in wild-caught and captive raccoons. Eimeria procyonis-like oocysts were seen in 15 of 15 captive raccoons. In 6 of 6 juvenile wild-caught raccoons examined at necropsy, endogenous coccidian stages were seen in the small intestine. Two types of schizonts (large and small) were identified. Large schizonts were up to 110 µm long, contained 10-μm-long merozoites, and were in crypt glandular epithelial cells. Smaller schizonts were 10 µm long, contained 5-μm-long merozoites, and were at the tips of the villi. Only a few gamonts and no oocysts were seen in sections. These stages were thought to be of E. procyonis.

Crepidostomum opeongoensis Caira, 1985 (Trematoda: Allocreadiidae) is redescribed from fish hosts Hiodon alosoides and Hiodon tergisus in southern Manitoba. The redescription adds details regarding the surface morphology and reproductive structures of the parasite not described previously. These include the characteristic tegumental papillae around the oral opening, paired papillae on the ventral and dorsal surfaces of the body, and tegumental bosses on the dorsal surface of the fore-body. Crepidostomum opeongoensis co-occurred with its hypothesized sister species, Crepidostomum illinoiense, in all host individuals harboring the former, and both species are characteristic of hiodontids. The association of C. opeongoensis with hiodontids and the absence of hiodontids in Lake Opeongo where C. opeongoensis was originally reported indicate that the Algonquin Park region may have had hiodontids in the past and that the life cycle may be completed at present without the original fish host.

Pterygodermatites (Mesopectines) senegalensis n. sp. (Nematoda: Rictulariidae) is described from Mastomys huberti in Senegal. It differs from other species of the subgenus by the morphology of the head, which exhibits 4 simple cephalic papillae and a nearly axial buccal opening, the length of the spicules, the number of precloacal cuticular formations, and the absence of cephalic apophyses.

Two new species of coelomic filarioid belonging to Litomosoides are described from sigmodontine murids from the Rio de La Plata marshland, Argentina. Litomosoides bonaerensis n. sp., a parasite of Oligoryzomys delticola, belongs to the carinii group and is close to L. silvai, which differs by the head and tail papillae, buccal capsule and cavity, area rugosa, and morphology of the microfilaria. Litomosoides oxymycteri n. sp., from Oxymycterus rufus, belongs to the sigmodontis group. Differential diagnosis is based on the morphology of the buccal capsule, the head and tail papillae, and microfilaria. The ectoparasitic gamasid Ornithonyssus bacoti, in which several Litomosoides species develop, has been recovered from sigmodontines trapped in the study.

The phylogenetic relationships of 14 species of Stilestrongylus were analyzed using the comparative morphology of 21 characters. We obtained 2 shortest trees of 50 steps, with a consistency index of 0.540 and 25 apomorphic character states. Ingroup monophyly was supported on these trees by 2 and 3 synapomorphies, respectively, and the ingroup was defined by the following characters: greater number of ventral ridges relative to the number of ridges dorsally, asymmetric bursa, and externodorsal rays differing in size. Sister-group relationships among Stilestrongylus and the other genera designated as outgroups are relatively consistent with those postulated by Durette-Desset's in a classification of the Nippostrongylinae. The ancestor of the species groups comprising Hassalstrongylus, Guerrerostrongylus, and Stilestrongylus originated and diversified in murids in the Neotropical region. Species of Hassalstrongylus occur in rodents between southeastern North America and the eastern part of South America (Brazil and Argentina), whereas species of Guerrerostrongylus and Stilestrongylus diversified exclusively in rodents in the Neotropics.

Parasites are often found in a milieu that requires extensive preparation and labor-intensive cleaning before they are suitable for use in analytical procedures. Application of modern techniques in immunology and molecular biology demands pure yields of parasites. To purify first-stage (L1) larvae of Elaphostrongylus cervi, fecal suspensions from an infected red deer were processed by the Baermann method and embedded in a gel matrix with the objective of selectively trapping fecal debris. About half the number (50.9%) of embedded larvae migrated out of the gel within a 24-hr period and were collected as clean parasite suspensions, virtually free from fecal debris. The numbers of L1 emigrating from gels were inversely proportional to the fecal debris content and the thickness of the gel. Removal of fecal debris from Baermann fluid by sieving prior to gel embedment enhanced the yield of pure L1.

Identification of the transmission routes of the trematode parasite Paragonimus miyazakii into different intermediate hosts would help to explain the natural distribution of the parasite. The behavior of P. miyazakii cercariae released from snails into water and in the presence of a living host or a whole crab leg was observed by stereoscopic or light microscopy at various times after exposure started. On encountering a crab leg or cheliped, the cercariae became entangled with the host via mucoid strands arising from the cercariae. Within 3 hr, most cercariae were attached to the host; cuticular penetration took between 5 and 6 hr, after which cercariae were found in the cavity of the leg. Crabs examined 102–149 days after exposure to the cercariae contained fully developed metacercariae. The metacercariae were fed to 2 rats, and the rats were killed 83 or 111 days later. Some of the metacercariae had reached maturity in the rats. That the cercariae were not ingested by the crabs but penetrated the crabs percutaneously (through hard as well as soft tissue) means that transmission can occur even in areas in which crabs and the host snails do not coexist, as they would if the usual route were oral (when the crabs ate infected snails).

The comparative susceptibilities of colonized species A, B, and C of Anopheles culicifacies complex and Anopheles stephensi were determined for 2 rodent malaria parasites Plasmodium vinckei petteri and Plasmodium yoelii yoelii. All the 3 members of the complex were found to support complete sporogony with varying success. Controls, A. stephensi, become readily infected, with >70% developing oocysts. Of the test groups, species A had the highest percentage of mosquitoes with oocysts (>25%) and sporozoites (>15%). Anopheles culicifacies species B were least susceptible; less than 10% had oocysts and sporozoites in the salivary glands. The results demonstrate that A. culicifacies species A is most susceptible and species B is least susceptible to infections with both the parasites.

Studies on several aspects of the life cycle of hookworms (Uncinaria spp.) in the California sea lion (Zalophus californianus) and northern fur seal (Callorhinus ursinus) were conducted on material collected on San Miguel Island (SMI), California and at The Marine Mammal Center, Sausalito, California in 1997, 1998, and 1999. Examination of Z. californianus intestines for adult hookworms and feces for eggs revealed that longevity of these parasites in pups is about 6–8 mo, and infections are probably not present in older sea lions. Parasitic third-stage larvae (L₃) were recovered from the ventral abdominal tissue of Z. californianus, suggesting transmammary transmission. Callorhinus ursinus pups had no hookworm eggs in their feces or adult worms (except for 1 probable contaminant) in their intestines in the fall and early winter, revealing that adult Uncinaria spp. are spontaneously lost at <3 mo of age of the pups. Sand samples from rookeries, used by both Z. californianus and C. ursinus, on SMI were negative for free-living, L₃ in summer months but positive in fall and winter months, indicating seasonality occurred.

The Cryptosporidium “human” genotype was identified in a paraffin-embedded tissue section from a dugong (Dugong dugon) by 2 independent laboratories. DNA sequencing and polymerase chain reaction/restriction fragment length polymorphism analysis of the 18S ribosomal RNA gene and the acetyl CoA synthethase gene clearly identified the genotype as that of the Cryptosporidium variant that infects humans. This is the first report of the human Cryptosporidium genotype in a nonprimate host.

Tritrichomonas foetus and Trichomonas vaginalis are protozoan parasites that cause sexually transmitted diseases in cattle and humans, respectively. There is a need for new antimicrobial agents to treat or prevent trichomoniasis because there are currently no approved chemotherapeutic agents against T. foetus and resistance of T. vaginalis to metronidazole does occur. Therefore, we evaluated the effect of a novel antimicrobial peptide, d-hecate, on the viability of 6 isolates of T. foetus and T. vaginalis in vitro. Tritrichomonas foetus and T. vaginalis were grown to mid log phase (24 hr) or late log/stationary phase (48 hr). Parasites at 10⁶/ml were mixed with equal volumes of d-hecate to final concentrations of 10 µM, 20 µM, and 40 µM of d-hecate. Controls had minimal essential medium (MEM) alone. The numbers of viable parasites were determined microscopically after 10, 20, and 30 min of incubation at 37 C with d-hecate or MEM. Our results show that d-hecate killed all 6 isolates of T. foetus and T. vaginalis evaluated. The killing effect was dependent on the concentration of the peptide, incubation time, and phase of growth of the parasites. Ultrastructural studies of parasites treated with 10 µM of d-hecate revealed extensive damage to the plasma membrane of most T. foetus and T. vaginalis cells, while a few cells were distorted but remained intact. d-Hecate may be a useful chemotherapeutic agent for the treatment of trichomoniasis.

Sanguinicola occidentalis (Trematoda: Sanguinicolidae) infected 58 (48%) of 120 yellow perch collected in 1997 and 40 (50%) of 80 yellow perch collected in 1998 from Silver Creek in lower Michigan. The mean intensity and mean abundance of this blood fluke were higher in 1998 than in 1997. The fluke was found in the bulbous arteriosus of the perch heart, free in the petri dishes when the gill filaments were teased apart, and in the body cavity washings, and 1 individual was associated with an eye. Of the 269 S. occidentalis examined from perch, none had eggs. Most perch examined and infected were only 1 yr in age. Spearman's correlation coefficients between S. occidentalis intensity and host length in 1997 and 1998 were not significant. An additional 25 yellow perch (0 yr in age) collected in 1998 were not infected. Thirty-seven (33%) of 113 snails (Campeloma decisum) examined in July and August 1999 from Silver Creek were infected with S. occidentalis.

Five murine monoclonal antibodies (MAbs) were developed against purified sporozoites of Hammondia hammondi. Despite a large antigenic similarity between the 2 closely related coccidia, H. hammondi and Toxoplasma gondii, these MAbs only reacted with H. hammondi. Three MAbs, 1D3, 3F2, and 4C9-7, recognized antigens of 38 kDa localized in rhoptries (1D3), in rhoptries and in oocyst and cyst walls (3F2), and in rhoptries and the apical region (4C9-7). Another MAb, 4C9-10, reacted with a 27-kDa antigen in dense granules of sporozoites and tachyzoites, and MAB 11B3 labeled an antigen of >94 kDa located in the pellicular membrane of the 3 stages of the parasite. These MAbs could be used for a rapid discrimination of the 2 coccidia in epidemiological studies or for diagnostic purposes in tissues.

Neospora caninum is a recently described coccidial parasite that was first isolated from a dog in 1988 and has subsequently been shown to infect a wide range of mammals. Neospora hughesi, a new species of this genus, has recently been isolated from the spinal cord of horses showing clinical signs of equine protozoal myeloencephalitis. The random amplified polymorphic DNA polymerase chain reaction technique is capable of differentiating between N. caninum and N. hughesi.

The primary structure of the ribosomal DNA internal transcribed spacers (ITS-1 and ITS-2) and 5.8S rRNA gene were used to characterize and identify 2 monogenean species of Gyrodactylus living externally on the threespine stickleback (Gasterosteus aculeatus). The ITS region was amplified by PCR from freshwater, brackish, and marine isolates of Gyrodactylus arcuatus and G. branchicus, and the ends of the coding regions were identified by comparative alignment. No intraspecific and very low interspecific variation were observed in the 5.8S rRNA gene; high inter- and low intraspecific variation were revealed in the ITS-1 and ITS-2 regions. The morphological species identification was in all cases confirmed by the molecular identification. Intraspecifically, samples from 2 locations in the North Sea could be differentiated, but the Baltic sample resembled North Sea genotypes. Our approach offers perspectives for a multimetric genetical, morphometrical, and ecological taxonomy of the genus Gyrodactylus.