Two superoxide dismutases (SOD I and SOD II) were purified from Acanthamoeba castellanii and characterized for several biochemical properties. Analysis of the primary structure and inhibition studies revealed that SOD I is iron SOD (Fe-SOD), with a molecular mass of 50 kDa, and SOD II is copper–zinc SOD (Cu,Zn-SOD), with a molecular mass of 38 kDa. Both enzymes have a homodimeric structure consisting of 2 identical subunits, each with a molecular mass of 26 and 19 kDa for SOD I and SOD II, respectively. The isoelectric points of SOD I and SOD II were 6.4 and 3.5, respectively, and there were no isoenzyme forms detected. Both enzymes show a broad optimal pH of 7.0–11.0. Because no differences were observed in the apparent molecular weight of SOD I after addition of the reducing agent 2-mercaptoethanol, the subunits do not appear to be linked covalently by disulfide bonds. However, the subunits of SOD II were covalently linked by intra- and interdisulfide bonds. Western blot analyses showed that the 2 enzymes have different antigenicity. Both enzymes occur as cytoplasmic and detergent-extractable fractions. These enzymes may be potential virulence factors of A. castellanii by acting both as antioxidants and antiinflammatory agents. These enzymes may be attractive targets for chemotherapy and immunodiagnosis of acanthamoebiasis.

The ability of Schistosoma mansoni to escape oxidative damage from immune system-generated reactive oxygen intermediates has been extensively documented. The limiting step in the parasite's detoxification process appears to be at the level of hydrogen peroxide neutralization. In the present study, the possible role of a novel class of antioxidant enzymes, thioredoxin peroxidase (TPx), in hydrogen peroxide neutralization by schistosomes was investigated. An expressed sequence tag was characterized from the Schistosoma Genome Initiative with high similarity to TPx from other organisms. The gene encodes a polypeptide containing 2 conserved active-site cysteines and flanking amino acids, and 60–70% identity with previously characterized TPx proteins. Recombinant schistosome TPx was enzymatically active and found to have thioredoxin-dependent hydrogen peroxide reducing activity of 4500 nmol hydrogen peroxide/min/mg protein. Native TPx activity was determined to be 48.1 nmol hydrogen peroxide/min/mg protein in adult worm homogenates compared with 46.9 for glutathione peroxidase. TPx activity was precipitated from adult worm homogenates with antibodies prepared against the recombinant protein. Western blotting with antibodies made against recombinant protein showed that TPx was expressed in both male and female adult worms. This is the first demonstration of a TPx activity in schistosomes and our results suggest that TPx plays a significant role in schistosome–host interactions.

The trichostrongylid nematode Haemonchus contortus released a hyaluronic acid–degrading enzyme during in vitro development from the third (L3) to fourth (L4) larval stage. The enzyme did not degrade chondroitin sulfate A. Enzyme activity was optimal between pH 4.0 and 6.0, and the enzyme was inhibited by high concentrations of NaCl; the divalent cations Cu² , Zn² , Ca² , and Mn² were not inhibitory. The hyaluronidase had a molecular mass estimated at 57 kDa by sucrose density gradient centrifugation and at 111 kDa by substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis (reducing and nonreducing conditions), suggesting the formation of a dimer during the electrophoretic separation conditions. The level of hyaluronidase released during in vitro development peaked between 24 and 48 hr in culture and then gradually decreased, with little or no activity present in the 168-hr culture fluid. The enzyme was not detected in culture fluid from 24-hr incubations of either the mid-L4 stage (obtained from sheep 7 days postinfection) or the adult stage (obtained from sheep 30–35 days postinfection). The temporal expression of the hyaluronidase suggested a role for this enzyme in the early stages of the L3–L4 developmental process.

The absence of a self-sustaining in vitro propagation method for Cryptosporidium parvum is a major obstacle for research on this parasite. Conventional cell monolayers are unsuitable for long-term parasite propagation because the level of infection decreases over time and few oocysts, if any, are produced. The interaction between parasite and host cell was studied to identify factors limiting parasite development in vitro. Loss of substrate adherence and death of parasitized host cells was observed in 2 epithelial cell lines. Nuclear morphology, DNA laddering, annexin V binding, and terminal deoxytransferase-mediated dUTP nick end labeling indicated that host cell death occurred by apoptosis. At 6 hr postinfection, only a minority of infected cells remained in the monolayer, and few survived the initial phase of parasite development without losing adherence. Treatment of infected monolayers with caspase inhibitors drastically reduced cell detachment but failed to increase the number of parasites in monolayers. In contrast, cell cultures grown on laminin-coated plates showed a higher proportion of infected cells. These observations indicate that cell detachment and apoptosis in C. parvum-infected cell culture negatively affect parasite survival in vitro.

The composition of the metazoan parasite communities within 2 fishing zones along the Chilean coast were compared to determine the population structure of the jack mackerel Trachurus symmetricus murphyi (Nichols) in these 2 geographical locations. More than 7,780 parasites belonging to 15 taxa were collected between 1990 and 1996 in 71 samples comprised of 3,946 hosts. The same taxa were found in both fishing zones. However, jack mackerel from northern Chile had a higher abundance of cymothoid isopods, Ceratothoa spp., whereas those from southern Chile had more Rhadinorhynchus trachuri, Hysterothylacium sp. larvae, and Anisakis type I larvae. Results were similar at all community levels because all parasites had low prevalence and infracommunities had low total abundance and richness. Analyses at the component community level may be more adequate for stock identification when infracommunities are simple. There were significant differences in composition of parasite communities between years in each fishing zone, presumably as a result of the increase in offshore catches since 1994. These results reinforce the hypothesis that more than 1 ecological stock of the jack mackerel exists in the southeastern Pacific and contradict the current assumption of a single stock in the management of this heavily exploited fish species.

Aggregation of Crataerina melbae flies on breeding adult alpine swifts (Apus melba) was low when compared with other host–parasite systems and varied with sampling date, year, and sex of the flies. Generalized linear models were performed to ascertain which factors, extrinsic and/or intrinsic to the host, explained variability in the number of louse flies present on a single host, i.e., abundance. Overall abundance was unrelated to any host characteristic but varied slightly among years. Abundance of female flies varied among years, but also with date of sampling, the number of females increasing as the breeding season advanced. In contrast, abundance of males decreased as the season progressed, independently of host characteristics. Despite these different patterns, the number of flies of each sex on a given host was strongly intercorrelated. These results suggest that mate attraction may explain aggregation patterns in this louse fly species. Overall sex ratio of louse flies did not differ from unity. However, the proportion of males decreased during the breeding season, as a consequence of the opposite sex-related seasonal patterns in parasite abundance. Sex-ratio variability was not related to host characteristics or to infrapopulation sizes.

Two new species of Calicotyle (Monocotylidae: Calicotylinae) are described from elasmobranchs in the western Gulf of California. Calicotyle californiensis n. sp. is described from a single specimen collected from a gray smoothhound shark (Mustelus californicus, Carcharhiniformes: Triakidae). It is distinguished from its congeners by the combination of having vaginal pores opening within the intercecal space, distal regions of the vaginae twisting, proximal regions of the vaginae fusing medially to form a kidney bean–shaped structure, and a relatively long male copulatory organ recurving 3 times and passing between the distal penis bulb and the seminal vesicle. Calicotyle urobati n. sp. is described from 16 specimens collected from at least the cloaca and rectum of the round rays Urobatis halleri and Urobatis maculatus (Rajiformes: Urolophidae). It is distinguished from its congeners by the combination of having vaginal pores opening outside the intercecal space and proximal regions of the vaginae terminating at the level of the ceca. Members of Calicotyle have not been reported previously from the eastern Pacific Ocean or from these hosts. In the past, species of Calicotyle have been distinguished based primarily on the shape and length of the male copulatory organ and hamuli. Divisions of the vaginae and the positions of the vaginal pores are also useful in distinguishing members of the genus.

The female of Ergasilus thatcheri n. sp. (Copepoda, Poecilostomatoida, Ergasilidae) is described from the gills of the “jundiá,” Rhamdia quelen (Teleostei, Siluriformes, Pimelodidae), from fish ponds in the State of Rio Grande do Sul, Brazil. The new species has a falciform semipinnate terminal seta on the first exopod, which suggests phylogenetic affinity to 10 other congeners, all from fishes of the Amazon Basin. Based on general morphology, the new species shows great similarity to E. callophysus Thatcher and Boeger, 1984, a parasite of another pimelodid fish Callophysus macropterus. Ergasilus thatcheri n. sp., however, can be easily differentiated from E. callophysus in its possession of a more distal sensillum not located on a cuticular elevation, and a row of spinules on the inner margin of the second antennal segment (first endopodal segment).

Campanulotes flavus (Rudow, 1869) from Phaps chalcoptera is redescribed and illustrated. C. defectus Tendeiro, 1969, erroneously recorded from the extinct passenger pigeon Ectopistes migratorius, is shown to be a junior synonym of C. flavus, thus removing it from the list of lice thought to have gone extinct with the passenger pigeon. C. flavus elegans (Tendeiro, 1978) from P. elegans is elevated to specific rank and redescribed. Finally, C. durdeni n. sp. is described from its type host Ocyphaps lophotes.

Chewing lice, Geomydoecus and Thomomydoecus, coexist on pocket gophers, Thomomys spp. We investigated the spatial distribution of the 2 genera on their hosts and explored possible mechanisms of resource partitioning by chewing lice. Chewing lice appear to partition available host resources spatially, with Geomydoecus occurring primarily on the lateral and dorsal regions of the host, and Thomomydoecus occurring primarily on the lateral and ventral regions. Although spatial partitioning of the host habitat is evident, it does not appear to be explained by hair diameter. Spatial partitioning of the host's body could be the result of some other factor, possibly temperature or humidity gradients of the host's body.

Canine red blood cell-substituted severe combined immune deficiency (Ca-RBC-SCID) mice were prepared for canine Babesia gibsoni infection. The Ca-RBC-SCID mice infected with B. gibsoni developed a high level of parasitemia, and showed clinical symptoms such as anemia and hemoglobinuria, which are similar to those observed in dogs infected with B. gibsoni. The B. gibsoni parasites grown in Ca-RBC-SCID mice showed marked morphological changes, including a significantly larger size of parasites than those in dogs and abundant RBCs containing 4, 8, 16, and 32 parasites. The multiple infection may have resulted from 1 parasite because the posterior end of each parasite in a multiply infected cell was connected. The parasites grown in SCID mice retained their infectivity and virulence to dogs and their morphology was dramatically restored to the original state when they were returned to dogs.

Molecular characterization of Toxoplasma gondii isolates is central for understanding differences in disease transmission and manifestations. Only 3 subgroups (lineages) have been discerned with subtle within-lineage variation, permitting low-resolution classification of isolates. Because proteins, coding sequences, and especially antigen-coding genes have been used extensively in previous studies, we focused on sequence variation in introns of housekeeping genes, which may be more informative for phylogenetic analysis because they evolve under lower selection. We compared sequence variation in introns of 5 housekeeping genes with 2 antigen-coding genes. Introns of housekeeping genes were slightly more polymorphic than coding and noncoding regions of antigen-coding genes and only the former showed intralineage variation. Intragenic linkage disequilibrium was complete, but intergenic linkage, although highly significant, was incomplete, suggesting that genes are partially uncoupled. Six of 7 substitutions found within the region coding for the tachyzoite surface antigen, SAG2, were nonsynonymous, indicating that diversifying selection acts on this locus. Typing isolates on the basis of housekeeping and antigen-coding genes was consistent, but the phylogenetic relationships among the resulting groups was inconsistent. A cougar isolate typed as lineage II using a restriction fragment length polymorphism assay possessed multiple unique polymorphisms, suggesting that it represents a new lineage. We concluded that introns of housekeeping genes are preferred markers for phylogenetic study, and that multilocus genotyping is preferred for typing parasites, especially from feral or unstudied environments.

The rRNA locus of Perkinsus atlanticus from the clam Ruditapes decussatus cultivated on the Atlantic coast of Spain was cloned and sequenced. Sequences of the internal transcribed spacer (ITS) from the rRNA locus were compared to sequences reported earlier for a P. atlanticus isolate from Portugal and to those from other Perkinsus species. The ITS1 sequence of the Spanish P. atlanticus isolate was identical to the Portuguese P. atlanticus sequence and had 76.6% identity to the ITS1 of Perkinsus marinus. The ITS2 sequence had 99.7% identity to the Portuguese P. atlanticus ITS2, 92.5% identity to the P. marinus ITS2, and 99.5% identity to the Perkinsus olseni ITS2. We report for first the time the small subunit (SSU) and nontranscribed spacer (NTS) of P. atlanticus. The P. atlanticus SSU sequence was 99.6% identical to that of an unidentified Perkinsus species from the Australian clam Anadara trapezia and 98.0% identical to that of P. marinus. Further, our results support the proposal that P. atlanticus, P. olseni, and the Perkinsus sp. from A. trapezia constitute a subgroup of Perkinsus species distributed in the Pacific and eastern Atlantic, different from P. marinus that is distributed along the western edge of the Atlantic. Based on the NTS sequence of P. atlanticus from Spain and the differences with P. marinus NTS (62.2% identity), we developed a polymerase chain reaction (PCR)-based diagnostic assay with a lowest limit of detection of 0.01 amol of cloned NTS DNA as assessed on ethidium bromide-stained agarose gels. Specificity of the PCR-based assay was tested with samples from the clams R. decussatus, Ruditapes philippinarum, and Venerupis pullastra collected in P. atlanticus-enzootic areas of Spain. The specificity and sensitivity demonstrated for this NTS-based PCR assay validate its use as a tool for assessment of P. atlanticus in molluscs.

Cryptosporidium parvum is an intracellular protozoan parasite of the mammalian intestine. In rats, C. parvum infection is age related; infants are susceptible, whereas adults are resistant. The transition from susceptibility to resistance usually takes place around the age of weaning. In the present study, infant rats were orally inoculated with a preparation of intestinal scrapings taken from adult rats or cows. Infant rats received the scrapings daily from 3 to 14 days of age, were inoculated with C. parvum oocysts at 9 days of age, and killed at 15 days of age. Fecal samples and intestinal tissues were examined for the presence of C. parvum. Significantly fewer rats were infected in the groups that received intestinal scrapings compared with controls. In addition, infected rats in the treatment groups shed significantly fewer oocysts than those in the control group. Scrapings from the intestinal mucosa of adult cows were also able to protect infant rats from infection, whereas scrapings from intestines of calves were not protective. In sum, these data indicate the presence of a factor in the intestines of adult rats and cows that can transfer protection against C. parvum infection to susceptible infant rats.

Immune responses to Cowdria ruminantium, an intracellular organism that causes heartwater in domestic ruminants, were characterized in a DBA/2 mouse model. Immunity induced by infection and treatment was adoptively transferable by splenocytes and could be abrogated by in vivo depletion of T cells but not by inhibition of nitric oxide synthase using N^G-monomethyl-l-arginine. IgG2_a and IgG2_b C. ruminantium-specific responses were detected in immune mice. Culture supernatants of splenocytes from immune DBA/2 mice, which were stimulated with crude C. ruminantium antigens or recombinant major antigenic proteins 1 or 2, contained significant levels of interferon (IFN)-γ and interleukin (IL)-6, but insignificant levels of IL-1α, IL-2, IL-4, IL-5, IL-10, IL-12, tumor necrosis factor-α (TNF), and nitric oxide. A similar response was detected during primary infection, although IFN-γ levels decreased significantly during clinical illness and then increased following natural or antibiotic-aided recovery. These data support the conclusion that protective immunity to C. ruminantium in DBA/2 mice is mediated by T cells and is associated with a polarized T helper 1 type of immune response. This murine model could be utilized to screen for protective C. ruminantium antigens that provoke Th1 type immune responses and for evaluation of these antigens in recombinant vaccines against heartwater.

The Schistosoma mansoni soluble adult worm antigen (SAWA) bands of 62/60 kDa were found to contain immunodominant T-cell immunogen(s) in irradiated cercariae-immunized Swiss and C57BL/6 mice. In the present study, spleen T cells of BALB/c mice immunized twice with ultraviolet light–irradiated cercariae proliferated and produced interleukin 4 in response to the 62/60-kDa SAWA bands in T-cell western assays. To characterize the 62/60-kDa bands, an adult S. mansoni worm cDNA expression library constructed in λgt11 was immunoscreened with serum of mice immunized with the 62/60-kDa antigens, and the immunoreactive cDNA inserts were sequenced. Purified 62- and 60-kDa proteins were used for amino acid microsequencing and for immunization studies in Swiss, C57BL/6, and BALB/c mice and rabbits. Taken together, the data indicated that the 60-kDa molecules are poorly immunogenic in mice and rabbits, whereas the 62-kDa species identified as S. mansoni calreticulin, is a good T- and B-cell antigen and represents a potential vaccine candidate.

We are attempting to design a simpler assay based on synthetic or recombinant antigens to replace the labor-intensive enzyme-linked immunoelectrotransfer blot (EITB-C), which is currently used to diagnose Taenia solium cysticercosis. From the lentil lectin-bound fraction of cyst glycoproteins (the LLGP fraction used in the EITB-C), we previously identified and purified 2 related polypeptides of 14- and 18-kDa that demonstrated diagnostic usefulness. Using degenerate oligonucleotide primers corresponding to amino acid sequences of these polypeptides and a cDNA library prepared from T. solium cysticerci, we amplified cDNA clones that represent the 14- and 18-kDa polypeptides. These clones share sequence homology at the nucleotide and amino acid levels. Synthetic polypeptides that represented the full-length, mature proteins (sTS14 and sTS18) were assessed for serologic potential using an ELISA. sTS14, but not sTS18, demonstrated utility as a diagnostic antigen. sTS14 was recognized by antibodies in a majority of the sera from patients with cysticercosis and none of the sera from persons with other helminth infections or uninfected human sera. Furthermore, polyclonal antibodies to sTS14 reacted with 6 discrete proteins present in the LLGP cyst fraction, suggesting that TS14 is a subunit of other previously described antigens used for diagnosing cysticercosis.

Unionicola poundsi and U. lasallei are recognized as closely related, morphologically distinct species of water mites living in symbiotic association with the mussels Villosa villosa and Uniomerus declivus, respectively. However, results of a transplant experiment suggested that the morphological characters used to separate these species are plastic and are influenced by the host species in which these mites metamorphose. These results indicate that U. poundsi and U. lasallei are variants of the same species. To test the validity of these contrasting notions, the genetic structure of mite populations from Uniomerus declivus and V. villosa was compared. An examination of allozyme variation at 9 enzyme loci revealed a high degree of genetic differentiation between these host-associated populations, with mites from U. declivus and V. villosa being fixed for different alleles at 3 loci and exhibiting significant allele heterogeneity at 71% of their polymorphic loci. Coefficients of genetic similarity and genetic distance for mites from U. declivus and V. villosa were 0.36 and 0.95, respectively. The results of this study suggest that mite populations from U. declivus and V. villosa are genetically distinct and complement morphological data recognizing them as valid species.

Digenetic trematodes usually show a high degree of specificity for their molluscan intermediate hosts. A panel of 4 digenean species (Echinostoma paraensei, E. trivolvis, Schistosoma mansoni, and Schistosomatium douthitti) and 5 snail species (Biomphalaria glabrata, Helisoma trivolvis, Lymnaea stagnalis, Stagnicola elodes, and Helix aspersa representing 3 gastropod families) was used to assess the relative contributions of miracidial behavior, host plasma osmolality, and host plasma factors in dictating specificity. Additional experiments were undertaken with a fifth digenean, Echinoparyphium sp. Expected patterns of compatibility were first confirmed; each parasite species produced patent infections in its known snail host, but not in the other snail species. One exception was S. douthitti, which unexpectedly did not infect L. stagnalis. As judged by direct observation and by noting their disappearance after exposure to snails, miracidia were generally less likely to attach to or penetrate incompatible than compatible hosts. However, over half of the miracidia of each parasite species attached to or attempted penetration of both compatible and incompatible hosts, suggesting that under the experimental conditions used, miracidial host location and attachment behaviors were not of overriding importance in dictating observed patterns of specificity. For each digenean species, the percentage of larvae that became immobile, rounded, showed tegumental damage, or died over a 6-hr interval in plasma of the various snails was assessed. In no case was plasma from a compatible host harmful to sporocysts or rediae. In contrast, in 8 of 16 (50 %) incompatible combinations, snail plasma had a significant negative effect on sporocyst condition. In 4 of 12 (33%) incompatible combinations, plasma had a significant negative effect on rediae. In 9 of 10 combinations tested, lymnaeid plasma was toxic for the parasites of planorbid snails and in 2 of 4 combinations, planorbid plasma was toxic for the parasites of lymnaeid snails. Toxicity was not attributable to differences in plasma osmolality between snail species. The ability of plasma from incompatible snails to affect viability of both sporocysts and rediae was surprisingly strong, suggesting that humoral factors play a greater role in dictating patterns of digenean–snail specificity than previously appreciated.

A panel of 4 digenetic trematode species (Echinostoma paraensei, E. trivolvis, Schistosoma mansoni, and Schistosomatium douthitti) and 5 snail species (Biomphalaria glabrata, Helisoma trivolvis, Lymnaea stagnalis, Stagnicola elodes, and Helix aspersa) was examined to determine if known patterns of host specificity could be explained by the tendency of digenean larvae to be bound by snail hemocytes, or by the ability of larvae to influence the spreading behavior of hemocytes. In short-term (1 hr) in vitro adherence assays, there was no overall pattern to suggest that sporocysts were more likely to be bound by hemocytes from incompatible than compatible snails. Compared with the other parasites, sporocysts of E. paraensei were less likely to be bound by hemocytes from any of the snail species tested. All rediae examined, including those of another species Echinoparyphium sp., were also remarkably refractory to binding by hemocytes from any of the snails. Of all the larvae examined, only sporocysts and young daughter rediae of E. paraensei caused hemocytes to round up in their presence. This was true for hemocytes from the compatible species B. glabrata and the incompatible lymnaeid species S. elodes and L. stagnalis. The patterns of host specificity shown by this particular panel of parasites and snails were not predicted by either the extent of hemocyte adherence to digenean larvae or by the ability of larvae to affect hemocyte spreading behavior. The results of this study suggest that a role for hemocytes, although likely, may require different assays, possibly of a more prolonged nature, for its detection. Also, different parasite species (notably E. paraensei) and intramolluscan stages have distinctive interactions with host hemocytes, suggesting that the determinants of specificity vary with the host–parasite combination, and with the parasite life cycle stage.

Balb/c mice were examined to determine whether or not they transmitted rodent malaria, Plasmodium berghei, to their fetuses. On the 15th day of pregnancy, mice were inoculated with approximately 3 × 10⁶ P. berghei-infected erythrocytes by peritoneal injection. The blood from 27 adult females and 196 neonates was examined using a sensitive polymerase chain reaction (PCR) method with a detection level of approximately 1 parasite/μl blood. The average parasitemia of females at delivery was 8.1%, ranging from nondetectable to 37.1%. In 12 females, nested PCR established the presence of blood parasite DNA. Malaria parasites were microscopically confirmed in 2 of the 12 neonates. Maternal parasitemia at the time of delivery was not correlated with the incidence of vertical infection (6.1%), which was higher in this study than that found in previous studies. Although the combination of balb/c mice and P. berghei has not been used to examine vertical transmission of malaria, our report showed that this model may be used for this purpose.

Blood smears of 1,353 free-ranging mammals (35 species) and 112 reptiles (31 species) from French Guiana were examined for hemoparasites. Parasites from 3 major groups were recorded: Apicomplexa (including hemogregarines, piroplasms, and Plasmodium spp.), Trypanosomatidae, and Filaroidea. Fifty percent of the individuals (86% of the species) were infected by parasites from at least 1 group. Hemogregarines, identified as Hepatozoon sp., infected numerous snakes with high prevalences (30–100%); infection is reported for the first time in 5 host genera of snakes: Clelia, Oxybelis, Pseustes, Rhinobotryum, and Bothriopsis. Infections were also observed in 4 marsupial species and 1 rodent. Hepatozoon spp. recorded in Didelphis albiventris (Marsupialia) and Coendou prehensilis (Rodentia) may be new species. Plasmodium sp. were observed in 2 snake species, Dipsas indica (Colubridae) and Bothrops atrox (Viperidae). Plasmodium brasilianum was recorded in all 5 primate species examined. Piroplasms were observed in all mammal orders except primates. Large terrestrial rodents were the main hosts of members of the Babesidae; 42% of Myoprocta acouchy, 36% of Dasyprocta agouti, and 44% of Agouti paca were infected. Trypanosomes were common in mammals and were recorded in 70% of the examined genera. Trypanosoma cruzi-like infections were reported in 21 mammal species, including sloths, rodents, carnivores, and primates. Microfilariae were also widespread, with higher prevalences in sloths, anteaters, and porcupines (>40% of the individuals infected) and in tamarins (95% infected). This survey highlights some potential anthropozoonotic risks due to the recent further evidence of Plasmodium brasilianum and P. malariae as a single species and to the increased diversity of hosts for Trypanosoma cruzi.

The course of infection of a malaria parasite (Plasmodium mexicanum) is highly variable in its host, the fence lizard (Sceloporus occidentalis). However, a seasonal trend is superimposed on this variation such that gametocyte production is intensified during mid- to late summer. Host testosterone levels follow a similar seasonal fluctuation and are variable among individual lizards. We sought to determine if testosterone levels affect seasonal and among-host variation in 11 P. mexicanum life history traits: rate of increase in level of infection (3 measures), peak parasitemia (3 measures), duration of increase (3 measures), time to detectable infection, and timing of production of gametocytes. We followed the course of infection in 125 male S. occidentalis, each randomly assigned to 1 of 4 treatment groups: castrated, castrated and implanted with exogenous testosterone, sham implanted, and unmanipulated controls. Median values for the 11 life history traits did not differ among treatment groups, and variances were homogeneous among the treatment groups for 10/11 traits. However, elevated testosterone significantly reduced the variation in timing of the onset of gametocyte production. Therefore, testosterone does not appear to be a primary regulator of P. mexicanum life history, yet testosterone may have some effect on when gametocytes first become detectable.

The life history of Ascarophis marina in Argentina is proposed on the basis of morphometric features of larval and adult specimens, as well as of trophic relationships among invertebrate and vertebrate hosts in the same geographic area. Adult and larval specimens of A. marina were found in the alimentary tract of the teleost fishes Parona signata (Carangidae) and Urophycis brasiliensis (Gadidae). Third-stage larvae ocurred in the shrimps Peisos petrunkevitchi (Sergestidae) and Artemesia longinaris (Penaeidae).

Southern flying squirrels (Glaucomys volans) from southeastern Georgia were examined for ectoparasites and gastrointestinal helminths. Ten species of ectoparasites were recovered, including 3 species of sucking lice (Hoplopleura trispinosa, Microphthirus uncinatus, and Neohaematopinus sciuropteri), 1 species of flea (Orchopeas howardi), 2 species of ticks (Amblyomma maculatum and Ixodes scapularis), 3 species of mesostigmatid mites (Androlaelaps casalis, A. fahrenholzi, and Haemogamasus ambulans), and 1 species of chigger (Leptotrombidium peromysci). Only the sucking lice and fleas were common on this host. M. uncinatus is reported for the first time from eastern North America. The 2 most commonly collected ectoparasites, N. sciuropteri (prevalence = 63%) and O. howardi (prevalence = 47%), have previously been shown to be vectors of the rickettsial zoonotic agent that causes sporadic epidemic typhus. Also, 3 nematodes (Citellinema bifurcatum, Strongyloides robustus, and Syphacia thompsoni), 1 unidentified cestode, and 1 acanthocephalan (Moniliformis clarki) were found in flying squirrel gastrointestinal tracts. With the exception of S. thompsoni, which was common and relatively abundant in the cecum (prevalence = 94%, intensity = 51 ± 12), both the prevalence and intensity of helminth parasites were low. The nematode S. thompsoni and the acanthocephalan M. clarki are new state records for tree squirrels in Georgia.

Intensity-dependent development of Ornithodiplostomum ptychocheilus metacercariae was studied in fathead minnows (Pimephales promelas) exposed to 0, 20, or 120 cercariae. Subsamples of hosts were necropsied at 2-wk intervals to monitor parasite recruitment, growth, and time to encystment. The complex development of metacercariae within the cranium of minnows involved growth, encystment, and consolidation phases, each of which were affected by intensity. At the end of the growth phase, metacercariae from low-dose fish were 20% longer than those from high-dose fish and the latter took 2–4 wk longer to encyst. At the end of the postencystment consolidation phase (6–8 wk postinfection), the size of metacercariae decreased by approximately 50%. The rate of consolidation was slower in high-dose fish. Our results show that time, intensity, and temperature affect development of O. ptychocheilus. Because metacercariae development and differentiation are linked to infectivity, events occurring in intermediate hosts can potentially impact the structure and size of trematode suprapopulations.

Aspects of the migratory life cycle and pathogenesis of Elaphostrongylus cervi were studied in red deer (Cervus elaphus) using 2 farmed calves experimentally infected with 450 third-stage larvae killed 40 and 45 days postinfection and using 3 wild calves and 3 wild yearlings with natural infections killed during autumn hunting. A full necropsy was carried out on the experimental calves, but only the head, eviscerated carcass, and lungs were examined from the naturally infected animals. Histological examination included extensive studies of the central nervous system (CNS), spinal nerve roots, and lungs. The experimental calves had prepatent infections, with many immature adult nematodes in the CNS, whereas the wild calves showed CNS lesions indicating a very recent E. cervi infection. The yearlings had patent infections, with many mature E. cervi in their skeletal muscles, reflecting acquisition of infection during the previous summer. Our findings showed that E. cervi develop to the adult stage in the CNS (subarachnoid spaces) and subsequently migrate into the skeletal muscles, where the mature nematodes live in reproductive pairs and groups. In the nervous system, the nematode caused encephalomyelitis, focal encephalomalacia and gliosis, meningitis, radiculitis, ganglionitis, and perineuritis.

Oxidative stress as a mediator of hepatic tissue damage concurrent with Leishmania (L.) chagasi infection was investigated. Chemiluminescence in liver supernatant of hamsters infected with Leishmania (L.) chagasi showed a ratio of 1.53/mg protein and 2.10/liver weight 90 days after infection when compared with the control. The malondialdehyde (MDA) levels also increased significantly both with and without addition of Fe³ /ascorbic acid in the reaction mixture, with a ratio of 2.12 and 1.55/mg protein or 2.91 and 2.12/liver weight, respectively. The parasite burden in the spleen, as a measure of infection severity, was 9.1 ± 1.33 × 10⁸ parasites/organ. On the 10th day of infection, the chemiluminescence also was significantly higher in infected hamsters than in the controls (ratio = 1.36/mg protein or 1.34/liver weight); however, the MDA levels were not different from those of controls. After 90 days of infection, significant correlations were observed between chemiluminescence and MDA concentration with and without the presence of Fe³ /ascorbic acid (r = 0.54, P = 0.0001; r = 0.56, P = 0.0001; respectively). The high infection/control ratio of both chemiluminescence and MDA concentration and the significant correlation between those events strongly indicate the occurrence of oxidative stress and lipid peroxidation as a mechanism of liver damage in cases of chronic infection by L. chagasi. The significant increase in chemiluminescence at 10 days of infection demonstrates that oxidative stress occurs very early, first consuming the antioxidants and then inducing lipid peroxidative damage later in the chronic stage of this disease.

Mice with targeted disruptions in the T-cell receptor α gene (TCRα^−/−) spontaneously develop inflammatory intestinal lesions with extensive B-cell lamina propria infiltrates. Cryptosporidium parvum infection accelerates intestinal lesion formation in TCRα^−/− mice. In the present study, TCRα^−/− mice were crossed with JH^−/− (B-cell-deficient) mice and challenged with C. parvum to determine if B cells are required for intestinal lesion development. TCRα^−/− × JH^−/− mice challenged with C. parvum, either as neonates or adults, became persistently infected, whereas TCRα^−/ × JH^−/ heterozygote control mice cleared the parasite. Cryptosporidium parvum colonization of TCRα^−/− × JH^−/− mice was heaviest in the distal ileum, with fewer parasites detected in the cecum and distal colon. Despite persistent infection, TCRα^−/− × JH^−/− mice did not develop inflammatory or hyperplastic intestinal lesions as detected in C. parvum-infected TCRα^−/− mice. These findings demonstrate that B cells are a necessary component for the development of inflammatory intestinal lesions of C. parvum-infected TCRα^−/− mice.

Ditrachybothridium piliformis is a new species from the spiral intestine of a cat shark, Galeus sp., from the southern Pacific Ocean. This is only the second species assigned to Ditrachybothridium. It differs from the type species D. macrocephalum in lacking spines on the scolex, a character originally used to diagnose the genus. The diagnoses of the Ditrachybothridiidae and of Ditrachybothridium have been revised to reflect this difference. This new species is further differentiated from the type species in its possession of pectinate spinitriches on the tegument of the scolex. The holdfast structures of this species are weakly muscularized, with no membrane-bound layer of radial muscles, indicating that the holdfast structures are bothria rather than bothridia as described in the most recent literature. Several reports for other species have indicated the same situation in other diphyllideans. The diagnosis of the order has been revised to reflect this finding.

A new species of Tylocephalum is described from the spiral intestine of the cownose ray Rhinoptera bonasus taken off Venezuela. Tylocephalum brooksi n. sp. is the third species described parasitizing this host species, along with T. pingue and T. bonasum. T. brooksi is similar to congeners in the possession of a scolex with a fungiform pars apicalis, craspedote segmentation, circummedullary vitellarium, external seminal vesicle, bilobed ovary, and medioventral uterus. However, this species differs from all previously described species of Tylocephalum in having a collar around the base of the pars apicalis densely covered by large digitiform processes (microtriches?), and an internal seminal vesicle. The internal seminal vesicle is a feature currently used to differentiate Tetragonocephalum from Tylocephalum. Its presence in T. brooksi indicates that it should be considered a specific instead of a generic character. An emended diagnosis of Tylocephalum is proposed excluding this feature, along with distribution of the testes in the preovarian field and circummedullary distribution of vitelline follicles. Species of Tylocephalum and Tetragonocephalum are reviewed on the basis of original descriptions, and 2 distinct morphological groups of species defined. Species of the Tetragonocephalum group possess a combination of an acraspedote strobila, segments much longer than wide, testes limited to the region anterior to the cirrus sac, vitellarium of lateral bands, conspicuously enlarged genital atrium and pore, cirrus sac dorsal to genital atrium, and bisaccate uterus. Species of the Tylocephalum group differ from Tetragonocephalum in possession of a craspedote strobila, segments more quadrangular, testes distributed throughout the preovarian field, inconspicuous genital atrium, uniformly saccate uterus, cirrus sac lateral to genital atrium, and probable circummedullary vitellaria in all species.

The pattern of longitudinal ridges (synlophe) on the external cuticular surface of trichostrongylid nematodes has been shown to be of value for distinguishing species and determining relationships among higher taxa. In the process of studying Mecistocirrus digitatus, the large stomach worm of bovids of Asia that has been imported and established in the Americas, we observed remarkably similar synlophe patterns to those described for 3 species of Haemonchus and to those we examined in a species of Ashworthius. In all 3 genera, the synlophe is absent from the posterior part of the body. Only in Haemonchus does the synlophe extend beyond midbody. In both M. digitatus and Ashworthius sidemi, the synlophe extends posteriorly only about ¼ of body length. In all 3 genera, the synlophe consists of about 30 ridges in the region of the esophagus with variation among species in specific areas, including additional pairs of subventral and subdorsal ridges and different lengths of sublateral ridges. This information is useful for identifying species and determining relationships among these large stomach worm parasites of cattle, sheep, goats, and farmed and wild cervids.

California sea lions (Zalophus californianus) and northern fur seals (Callorhinus ursinus) are each believed to host distinct hookworm species (Uncinaria spp.). However, a recent morphometric analysis suggested that a single species parasitizes multiple pinniped hosts, and that the observed differences are host-induced. To explore the systematics of these hookworms and test these competing hypotheses, we obtained nucleotide sequences of nuclear ribosomal DNA (D2/D3 28S, D18/D19 28S, and internal transcribed spacer [ITS] regions) from 20 individual hookworms parasitizing California sea lion and northern fur seal pups where their breeding grounds are sympatric. Five individuals from an allopatric population of California sea lions were also sampled for ITS-1 and D18/D19 28S sequences. The 28S D2/D3 sequences showed no diagnostic differences among hookworms sampled from individual sea lions and fur seals, whereas the 28S D18/D19 sequences had one derived (apomorphic) character demarcating hookworms from northern fur seals. ITS sequences were variable for 7 characters, with 4 derived (apomorphic) states in ITS-1 demarcating hookworms from California sea lions. Multivariate analysis of morphometric data also revealed significant differences between nematodes representing these 2 host-associated lineages. These results indicate that these hookworms represent 2 species that are not distributed indiscriminately between these host species, but instead exhibit host fidelity, evolving independently with each respective host species. This evolutionary approach to analyzing sequence data for species delimitation is contrasted with similarity-based methods that have been applied to numerous diagnostic studies of nematode parasites.

During a long-term study carried out between 1981 and 1996 on the biological and fishery aspects of octopuses inhabiting the Gulfs of San Matías, San José, and Nuevo, Patagonia, Argentina, 2 new species of Aggregata (Apicomplexa: Aggregatidae) were found in the digestive tracts of Octopus tehuelchus d'Orbigny, 1834 (prevalence 72%) and Enteroctopus megalocyathus (Gould, 1852) (prevalence 77%). Both species can be distinguished from other congeners on the basis of their hosts, the diameters of sporocysts, and number and length of sporozoites. Despite overlap in the distributions of the two hosts species in the area covered in this study, both Aggregata new species exhibited high host specificity.

Halipegus eschi n. sp. is described from the esophagus of Rana vaillanti from Guanacaste Province, Costa Rica. The new species differs from other known species of Halipegus on the basis of relative testis size, lateral extent of the uterus, vitelline follicle arrangement, egg size, and polar filament length.

An unexpected activity of β-cyclodextrin, an excipient used in pharmaceutical technology, was observed against Cryptosporidium parvum. The viability and infectivity of purified oocysts, exposed for 24 hr to β-cyclodextrin (2.5% suspension), were evaluated by inclusion/exclusion of 2 fluorogenic vital dyes and a suckling murine model, respectively. Results of the viability assay showed a high proportion of nonviable oocysts (81.5%). The intensity of experimental infection, determined 7 days postinoculation by examination of intestinal homogenates, was significantly lower (P < 0.05) than in the control litters. The preventive and curative efficacies of β-cyclodextrin suspension were also evaluated in experimentally infected neonatal mice. Infection was prevented when the suspension was administered 2 hr before inoculated oocysts and on days 1 and 2 postinoculation.

An immunoelectrotransfer blot assay (IETB) using excretory–secretory products of muscle larvae of Trichinella spiralis (ML-ESP) and the avidin–biotin system was developed in order to characterize reactivity against ML-ESP in sera from patients with acute and chronic trichinellosis. A complete pattern of up to 13 bands was developed by sera from individuals with trichinellosis where doublets, triplets, or single bands were shown to have molecular weights of roughly 66, 55, 45, 36, 29, 24, and 14 kDa. The bands at ∼55, 36, 29, and 14 kDa proved specific for T. spiralis. The band at ∼55 kDa was present in all trichinellosis sera, whereas the ∼14-kDa band was present in only a small percentage of sera. The development of ∼36- and 29-kDa bands suggests a modulation of the reactivity against ML-ESP over time. IETB proved more sensitive for the population of chronic trichinellosis under study than a conventional diagnostic enzyme-linked immunosorbent assay, allowing negative or borderline serum samples to be determined. Thus, this technique, when applied for human trichinellosis surveillance, should provide a useful tool in endemic areas.

A detailed temporal examination was made of alterations induced by artemether in the tegument of adult Schistosoma mansoni worms using scanning electron microscopy (SEM). Mice infected with S. mansoni cercariae 42 days previously were treated intragastrically with artemether at a single dose of 400 mg/kg. Groups of 3 mice were killed at 24 hr, 72 hr, and 7 days after treatment; the worms were collected by perfusion and examined by SEM. Twenty-four hours after artemether treatment, focal damage to the tubercles on the tegumental surface of male worms was seen. In both male and female worms, there was focal swelling and fusion of tegumental ridges, and sometimes peeling. After 72 hr, the damage to the tegument had increased, especially in female worms, with extensive swelling, fusion, and peeling of the tegumental ridges. In the most severely damaged worms, host leukocytes were seen to be adhered to the damaged tegument. Damage to the oral sucker was also occasionally seen in both male and female worms. Seven days after treatment, the appearance of the tegument had returned to normal in some male and female worms, whereas others still showed apparent damage. The results demonstrate that artemether damages the tegument of adult S. mansoni, and the intensity of damage is more severe in female worms than in males.

To elucidate the pathogenic potential of Toxocara cati, we observed the ophthalmologic changes of the fundi in Mongolian gerbils, Meriones unguiculatus, after oral inoculation of 17 embryonated eggs/g body weight. Ophthalmic conditions in 8 T. cati-infected gerbils were monitored using an ophthalmoscope from day 0 to day 156 and were compared with those of 57 T. canis-infected gerbils. The results showed that T. cati larvae migrated into the eye of the gerbil and then elicited ophthalmic changes, including retinal (25%) and vitreous (50%) hemorrhaging, vasculitis (37.5%), and exudative lesions (25%). Lesions were less prevalent, however, in T. cati-infected than in T. canis-infected gerbils. Unlike in T. canis-infected gerbils, the hemorrhagic lesions did not reappear in T. cati-infected gerbils after they were absorbed. These findings suggested that T. cati larvae are a potentially hazardous pathogen for ocular toxocariasis and that Mongolian gerbils infected with T. cati may be a useful model for the study of human ocular toxocariasis caused by T. cati. This is the first study to report that T. cati larvae can induce ophthalmic lesions in the retina of gerbils.

Amblyomma marmoreum and A. sparsum ticks were collected from tortoises imported into Florida from Africa and were tested for Cowdria ruminantium infection using a C. ruminantium-specific pCS20 polymerase chain reaction assay. In 1 shipment imported from Zambia, 15 of the 38 A. sparsum male ticks collected from the leopard tortoises (Geochelone pardalis) were found to be positive for infection with C. ruminantium. In contrast, all 148 A. marmoreum tested were negative for C. ruminantium infection. This is the first reported evidence of the introduction of heartwater-infected ticks into the United States, but there were no opportunities to confirm isolation of C. ruminantium from the ticks by either culture or transmission studies.

Schistosoma mansoni egg antigens are mostly responsible for the granulomatous pathology in human intestinal schistosomiasis. Several previous studies have indicated that the induction of an immune response against some parasite enzymes may protect against pathology. The present work was designed to identify enzyme activities present in a standard soluble egg antigen (SEA) preparation. Simple colorimetric analyses were performed incubating SEA with 2-naphthyl, 2-naphthylamide (2NA), or p-nitrophenyl substrates at different pHs in the absence of added effectors. Results showed prominent acid phosphatase (pH 5.4), alkaline phosphatase (pH 8.5), and N-acetyl-β-glucosaminidase (pH 5.4) activities. Relevant peptidase activities were also detected at pH 6.5–7.5 against 2NA derivatives of (1) aliphatic (α-Ala > β-Ala > Leu > Met > S-benzyl-Cys), polar (Ser > Gln), basic (Arg > Lys > ornithine), and acidic (Glu) amino acids; (2) dipeptides: X-Ala (X = Gly > Leu > Lys > Asp), X-Arg (X = Ala > Arg > Phe > Gly > Pro > Asp), Ser-Met, and Phe-Pro; and (3) tripeptides (Ala-Phe-Pro > Phe-Pro-Ala). The data demonstrated that S. mansoni SEA contains a rich set of hydrolases with different specificities that might play a role in the egg physiology and possibly also in the host–parasite relationships.

The genetic influence on host susceptibility to Neodiplostomum seoulense infection and fatality of the host was studied in 3 inbred strains of mice (BALB/c [H-2d], C3H/He [H-2k], and C57BL/6 [H-2b]). The survival of the mice, worm expulsion kinetics, worm size, number of eggs produced per day (EPD), and number of uterine eggs were observed from day 1 to day 40 postinfection (PI) with 100 or 200 metacercariae per mouse. Infection with N. seoulense was highly lethal to all 3 strains of mice, but the lethality was dose-dependent and varied according to the genetic backgrounds of the mice. The C3H/He mice exhibited the highest mortality, the lowest worm burdens and EPD, and the quickest expulsion of worms. It is suggested that different genetic backgrounds of mice appear to affect the host's capacity to expel N. seoulense and the fatality of the hosts themselves.

IR162 is a rat monoclonal IgE antibody (mAb). In the investigation of rat IgE production, the antigens recognized by IR162 (IR-Ags) were found to be expressed by a variety of helminthic parasites. By western blot analysis, IR162 detected bands in crude extracts of Anisakis simplex larvae, Clonorchis sinensis, Paragonimus westermani, plerocercoids of Spirometra mansoni, and Toxocara canis. Excretory–secretory material from A. simplex larvae also contained a protein recognized by IR162. IR162 mAb obtained from both Serotec and Zymed recognized identical bands of A. simplex larvae. Sandwich enzyme-linked immunosorbent assay analysis indicated that both IR-Ags of A. simplex larvae (IR-As) and C. sinensis (IR-Cs) were important antigens with respect to induction of specific IgG in human infections with these 2 organisms. Even though minimal cross-reaction between IR-As and IR-Cs was observed using sera from infected individuals, these results indicate that IR-Ags are found in the parasites examined, that IR-As and IR-Cs are the antigens that induce specific antibody in natural infection, and that the epitope that binds IR162 is shared by A. simplex larvae and C. sinensis.

A Frenkelia microti tissue cyst was found in the brain of a chinchilla (Chinchilla laniger) used for biomedical research. This is the first report of Frenkelia infection in this animal in the United States.

In the central nervous system of 2 raccoons, 1 cat, 1 pony, 2 mink, and 1 skunk, protozoa previously thought to be Sarcocystis-like reacted positively to Sarcocystis neurona-specific antibodies in an immunohistochemical test. In addition, S. neurona was identified in the brain of another skunk. These observations indicate that S. neurona is not confined to opossums and horses.

A species of Gyrodactylus collected from 2 species of anguillid eels (Anguilla australis Richardson, 1841 and Anguilla reinhardtii Steindachner, 1867) from Australia is identified as Gyrodactylus anguillae Ergens, 1960. The morphology of sclerites of G. anguillae specimens from Australia is in accordance with previous descriptions of specimens collected from A. anguilla (Linnaeus, 1758) from Europe and A. anguilla imported into Japan. Gyrodactylus anguillae was previously thought to be a parasite specific to A. anguilla, an eel that is native to freshwater catchments throughout Europe. Information on eel translocations and host and parasite biology is reviewed and it is hypothesized that G. anguillae is a naturally occurring parasite in Australia and not an introduction.

From June 1993 through June 1996, 2,260 adult, 4,426 nymphal, and 2,178 larval lone star ticks Amblyomma americanum (L.) were collected in Missouri from vertebrate hosts and by dragging a cloth over vegetation. Prevalence, mean intensity, and relative abundance of each stage varied among hosts. The relative abundance of adult lone star ticks was highest on white-tailed deer, but this stage was also collected from raccoons, opossum, red fox, coyotes, and wild turkey. Nymphs were collected from gray squirrels, eastern cottontail rabbits, opossums, red fox, Carolina wren, and bobwhite quail, but the highest relative abundance occurred on wild turkey, white-tailed deer, and raccoons. Eastern cottontail rabbits, white-tailed deer, raccoons, and squirrels had the highest relative abundance of larval lone star ticks, but they were also found on opossums and wild turkey. The activity of adult lone star ticks was greatest from May through July. The activity for nymphs was highest from May through August, and for larvae, July through September.

Grossly visible sarcocysts were seen in the skeletal muscles of 1 of 12 13-lined ground squirrels, Spermophilus tridecemlineatus tridecemlineatus, collected in Nebraska. The tissue cyst wall was up to 5.0 μm thick and contained spikelike projections. Transmission electron microscopy of tissue cysts revealed they were similar to Sarcocystis campestris Cawthorn, Wobeser, and Gajadhar, 1983, previously known only from experimental infections in Richardson's ground squirrel Spermophilus richardsonii. Prominent electron-dense bodies were observed lining the microfilaments present in the spikelike projections of the sarcocyst wall. This is the first report of S. campestris in a natural intermediate host and the first report of this parasite outside of Saskatoon, Canada.

High-performance thin-layer chromatography was used to analyze the neutral lipids in the rediae, cercariae, and encysted metacercariae of the paramphistomid trematode Zygocotyle lunata. Visual observations of the chromatograms showed that the most abundant lipid fractions were free sterols and free fatty acids in all larval stages and triacylglycerols in the metacercariae and rediae. The weight of free sterols (x̄ ± SE) was 120 ± 20 ng/cercaria, 56 ± 3.8 ng/redia, and 5.9 ± 1.5 ng/encysted metacercaria; the weight of triacylglycerols was 13 ± 0.88 ng/encysted metacercaria, 6.3 ± 0.063 ng/redia, and was not detectable in the cercaria; the weight of free fatty acids was 160 ± 17 ng/cercaria, 76 ± 9.1 ng/redia, and 4.2 ± 0.46 ng/encysted metacercaria. Oil red O staining of whole larvae showed the presence of neutral lipids in the rediae but not in the cercariae or encysted metacercariae. A dramatic reduction was seen in the quantity of free sterols and free fatty acids in the encysted metacercariae as compared with the cercariae, suggesting that these neutral lipids are used in some way during the transformation from cercaria to metacercaria.

Specimens of Thubunaea dactyluris Karve, 1938 sensu Fabio and Rolas, 1974 from the lizard Ameiva ameiva of Brazil and types of Physalopteroides venancioi (Lent, Freitas, and Proença, 1946) from the toad Bufo paracnemis of Paraguay were re-examined. The male caudal papillae and the tooth arrangement demonstrated that these specimens were conspecific and are all assigned to P. venancioi. The occurrence of Physalopteroides dactyluris (Karve, 1938), a nematode parasite of lizards of India and Turkmenistan, is invalidated for Brazil.

To further validate the observation of the existence of host-adapted strains of Cryptosporidium parvum, we genetically characterized an isolate of Cryptosporidium parasite from a black bear. Sequence analysis of the ribosomal RNA small subunit and the 70-kDa heat shock protein (HSP70) showed that this parasite represents a new genotype of C. parvum and is related to the C. parvum dog genotype. This finding is helpful for clarifying Cryptosporidium taxonomy.

Genomic DNAs from 3 species of Metagonimus in the Republic of Korea, M. yokogawai, M. miyatai, and M. takahashii, were amplified by simple sequence repeat anchored polymerase chain reaction. Primers of 3′ or 5′ termini of the (CA)_n repeats were used; 3 kinds of 3′ anchored primers (CA)₈RG, (CA)₄RG, and (CA)₈RY, and 2 kinds of 5′ anchored primers BDB(CA)₇C and DBDA(CA)₇. The results showed distinguishable banding patterns among the 3 species of Metagonimus, suggesting that they have different genotypes.