Cysticerci of Taenia crassiceps reproduce asexually by exogenous budding in the rodent intermediate host, and can experimentally develop to the adult stage within the small intestine of golden hamsters. In the present study, we report the loss of cysticercus infectivity for hamsters after maintaining the strain for 4 yr by consecutive peritoneal passage in mice. Larval infectivity was restored after a cysticercus from the WFU strain developed into a gravid tapeworm after being passaged through a dog. The eggs of this tapeworm were infective for mice, which subsequently developed cysticerci with renewed capability for infecting experimental hamsters. An in vitro evagination assay was also conducted using eleventh-generation WFU strain cysticerci, as well as second- and fourth-generation dog-derived cysticerci. Significantly higher (P < 0.0001) evagination was observed for 5-mo-old dog-derived and WFU infrapopulations when compared with respective evagination values for 9- and 12-mo-old infrapopulations. The extent of evagination was linked to the capacity of cysticerci to infect hamsters, so that greater evagination resulted in a higher infectivity for hamsters and vice versa.

The degree to which parasites use hosts is fundamental to host–parasite coevolution studies, yet difficult to assess and interpret in an evolutionary manner. Previous assessments of parasitism in eugregarine–host systems suggest high degrees of host specificity to particular host stages and host species; however, rarely have the evolutionary constraints on host specificity been studied experimentally. A series of experimental infections were conducted to determine the extent of host stadium specificity (larval vs. adult stage) and host specificity among 6 tenebrionid host species and 5 eugregarine parasite species. Eugregarines from all host species infected both the larva and adult stages of the host, and each parasite taxa colonized several host species (Tribolium spp. and Palorus subdepressus). Parasite infection patterns were not congruent with host phylogeny, suggesting that host phylogeny is not a significant predictor of host–parasite interactions in this system. However, the 2 host stages produced significantly different numbers of parasite propagules, indicating that ecological factors may be important determinants of host specificity in this host–parasite system. While field infections reflect extant natural infection patterns of parasites, experimental infections can demonstrate potential host–parasite interactions, which aids in identifying factors that may be significant in shaping future host–parasite interactions.

Parasite community structure can change seasonally with shifts in host habitat and in diet. However, anthropogenic activity may influence the natural changes in transmission dynamics of different parasite species. Effects of seasonal and agricultural activity on the parasite communities of newly metamorphosed northern leopard frogs (Rana pipiens) were investigated in July and September 2001 in 5 wetlands, 3 of which were exposed to pesticide runoff from surrounding agriculture. Nineteen parasite taxa were found. Component community richness was consistently high at the pristine reference wetland, whereas the communities at a managed reference wetland remained depauperate. Infracommunity richness increased throughout the season, but more so in frogs resident in agricultural wetlands. Digeneans using frogs as intermediate hosts dominated the communities, although many species were much lower in abundance in September, suggesting mortality of heavily infected frogs. Mean abundance of Haematoloechus spp. was positively related to that of odonate naiads in the frog diet, which appeared to reflect differential second intermediate host availability between reference and agricultural wetlands. Although virtually absent from wetlands in July just after frog metamorphosis, monoxenous nematodes were more prevalent and abundant at agricultural wetlands as the season progressed. Our results suggest that agricultural activity may further facilitate the transmission of monoxenous nematodes as frogs become more terrestrial.

The parasitic trematode Proctoeces lintoni requires 3 hosts (intertidal mussels, keyhole limpets, and clingfish) to complete its life cycle. The densities and size structure of host communities are modified by selective human harvesting. This study examined clutch and egg size of P. lintoni in 3 adjacent sites in rocky intertidal areas of central Chile presenting differences in the levels of human disturbance (i.e., from a fully protected marine reserve to free open-access areas). We found significant differences in parasite fecundity among sites. An increase in number of eggs was observed inside protected marine areas compared with open-access areas, suggesting a plastic response of the parasite reproductive strategies to the host community modification. These results show that host removal by humans in coastal ecosystems can strongly influence parasite life history traits.

In the course of the investigations into the fish parasites in the Tres Palos Lagoon in the State of Guerrero off the Pacific coast of Mexico, the following diplectanid species (Monogenoidea) from the gills of centropomids were found: Rhabdosynochus alterinstitus n. sp. from Centropomus nigrescens; Rhabdosynochus lituparvus n. sp., Rhabdosynochus volucris n. sp., and Rhabdosynochus siliquaus n. sp. from Centropomus robalito (Centropomidae). The apparent synapomorphic character supporting a sister relationship of these diplectanids is a single, sheathlike accessory piece comprising 3 distal branches of the male copulatory organ. The origin of the present diplectanid on centropomids is discussed, and it is suggested that this may be the result of allopatric speciation as a result of the uplift of the Panamanian Isthmus, thereby separating the Pacific and Atlantic Oceans during Pleistocene (3–5 million yr ago).

The prevalence of Toxoplasma gondii was investigated on a poorly managed pig farm in Maryland. Serum and tissue samples from 48 of the 100 pigs on the farm were available for T. gondii evaluation. Serological testing was performed using both ELISA and the modified agglutination test (MAT). Antibodies to T. gondii were detected by ELISA in 12 of 48 animals, while antibodies were detected in 34 of 48 pigs by MAT with titers of 1:10 in 1, 1:20 in 4, 1:40 in 7, 1:80 in 3, 1:160 in 8, 1:320 in 3, 1:640 in 4, and 1:1,280 in 4. Hearts of 16 pigs with MAT titers of 1:10 or higher were bioassayed for T. gondii in cats; 11 cats shed T. gondii oocysts. Hearts of 22 pigs were autolyzed and bioassayed only in mice; T. gondii was isolated from 3 of these 22 pigs. Genetic typing of the 14 T. gondii isolates using the SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico loci revealed 4 genotypes; 10 isolates belonged to type II lineage (genotypes 1 and 2), 3 belonged to genotype 3, and 1 belonged to genotype 4. Genotype 1 and 2 have type II alleles at all genetic loci, except the former has type II allele and the latter has a type I allele at locus Apico. Both genotypes 1 and 2 are considered to belong to the clonal type II lineages. Genotype 3 and 4 are nonclonal isolates. Results document high prevalence of T. gondii in pigs on a farm in Maryland.

Viable Toxoplasma gondii was isolated by bioassay in mice from tissues of 2 feral cats (Felis domesticus), 2 raccoons (Procyon lotor), a skunk (Mephitis mephitis) trapped in remote locations in Manitoba, Canada, and a black bear (Ursus americanus) from Kuujjuaq, northern Quebec, Canada. Genotyping of these T. gondii isolates using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico revealed 4 genotypes. None of the isolates was clonal archetypal Types I, II, and III found in the United States. These results are in contrast with the Type II genotype that is widespread in domestic animals and humans throughout the United States and Europe. This is the first genotyping of T. gondii isolates from this part of North America.

Henneguya gurlei was isolated from Ameiurus nebulosus captured in North Carolina and redescribed using critical morphological features and 18S small-subunit ribosomal RNA (SSU rDNA) gene sequence. Plasmodia are white, spherical, or subspherical, occur in clusters, measure up to 1.8 mm in length, and are located on the dorsal, pectoral, and anal fins. Histologically, plasmodia are located in the dermis and subdermally, and the larger cysts disrupt the melanocyte pigment layer. The spore body is lanceolate, 18.2 ± 0.3 μm (range 15.7–20.3) in length, and 5.4 ± 0.1 μm (range 3.8–6.1) in width in valvular view. The caudal appendages are 41.1 ± 1.1 μm (range 34.0–49.7) in length. Polar capsules are pyriform and of unequal size. The longer polar capsule measures 6.2 ± 0.1 μm (range 5.48–7.06), while the shorter is 5.7 ± 0.1 μm (range 4.8–6.4) in length. Polar capsule width is 1.2 ± 0.03 μm (range 1.0–1.54). The total length of the spore is 60.9 ± 1.2 μm (range 48.7–68.5). Morphologically, this species is similar to other species of Henneguya that are known to infect ictalurids. Based on SSU rDNA sequences, this species is most closely related to H. exilis and H. ictaluri, which infect Ictalurus punctatus.

Complete sequences of ribosomal and mitochondrial genes of the giant liver fluke Fascioloides magna are presented. In particular, small subunit (18S) and internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene (rDNA), as well as cytochrome c oxidase subunit I (cox1) and nicotinamide dehydrogenase subunit I (nad1) of the mitochondrial DNA (mtDNA), were analyzed. The 18S and ITS sequences were compared with previously published sequences of the liver fluke Fasciola hepatica. Fixed interspecific genetic differences were determined that allow molecular differentiation of F. magna and F. hepatica using either the PCR-RFLP method or PCR amplification of species-specific DNA regions. Additionally, intraspecific sequence polymorphism of the complete cox1 and nad1 mitochondrial genes in geographically distinct F. magna populations was determined. Based on the sequence divergences, short (<500 bp) variable regions suitable for broader biogeographical studies of giant liver fluke were designed.

The prevalence of Toxoplasma gondii in free-ranging chickens (Gallus domesticus) is a good indicator of the prevalence of the parasite's oocysts in soil because chicken feed from the ground. The prevalence of T. gondii in free-range chickens from Ghana, Indonesia, Italy, Poland, and Vietnam was determined using the modified agglutination test (MAT). Antibodies to T. gondii were found in 41 (64%) of 64 chickens from Ghana, 24 (24.4%) of 98 chickens from Indonesia, 10 (12.5%) of 80 chickens from Italy, 6 (30%) of 20 chickens from Poland, and 81 (24.2%) of 330 chickens from Vietnam. Hearts and brains of chickens were bioassayed for T. gondii. Viable T. gondii was isolated from 2 chickens from Ghana, 1 chicken from Indonesia, 3 chickens from Italy, 2 chickens from Poland, and 1 chicken from Vietnam. Toxoplasma gondii isolates from 9 chickens were genotyped using 10 PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. A total of 7 genotypes was identified; the 3 isolates from chickens from Italy were clonal type II, and the others were nonclonal. This is the first report of genetic characterization of T. gondii isolates from animals from these countries.

Toxoplasma gondii is a protozoan pathogen of birds and mammals, including humans. The infective stage, the bradyzoite, lives within cysts, which occur predominantly in cells of the central nervous system and skeletal and cardiac muscles, characterizing the chronic phase of toxoplasmosis. In the present study, we employed for the first time primary mouse culture of skeletal muscle cells (SkMC) infected with bradyzoites, as a cellular model for cystogenesis. The interconversion of bradyzoite and tachyzoite was analyzed by immunofluorescence using 2 stage-specific antibodies, i.e., anti-bradyzoite (anti-BAG1) and anti-tachyzoite (anti-SAG1). After 24 hr of interaction only bradyzoites were multiplying, as revealed by anti-BAG1 incubation; interconversion to tachyzoites was not observed. After 48 hr of infection, 2 types of vacuoles were seen, i.e., BAG1 and SAG1 , indicating the presence of bradyzoites as well as their interconversion to tachyzoites. After 96 hr of infection, BAG1 vacuoles presented a higher number of parasites when compared to 48 hr, indicating multiplication of bradyzoites without interconversion. Using ultrastructural analysis, bradyzoites were found to adhere to the cell membranes via both the apical and posterior regions or were associated with SkMC membrane expansions. During bradyzoite invasion of SkMC, migration of the rough endoplasmic reticulum (RER) profiles to the parasite invasion site was observed. Later, RER profiles were localized between the mitochondria and parasitophorous vacuole membrane (PVM) that contained the parasite. After 31 days of parasite-host cell infection, RER profiles and mitochondria were not observed in association with the cyst wall. Alterations of the PVM, including increased thickness and electrondensity gain on its inner membrane face, were observed 48 hr after infection. Cystogenesis was complete 96 hr after infection, resulting in the formation of the cyst wall, which displayed numerous membrane invaginations. In addition, an electron-dense granular region enriched with vesicles and tubules was present, as well as numerous intracystic bradyzoites. These results show that the in vitro T. gondii model and SkMC are potential tools for both the study of cystogenesis using molecular approaches and the drug screening action on tissue cysts and bradyzoites.

To establish and persist within a host, Leishmania spp. parasites delay the onset of cell-mediated immunity by suppressing interleukin-12 (IL-12) production from host macrophages. Although it is established that Leishmania spp.-infected macrophages have impaired IL-12 production, the mechanisms that account for this suppression remain to be completely elucidated. Using a luciferase reporter assay assessing IL-12 transcription, we report here that Leishmania major, Leishmania donovani, and Leishmania chagasi inhibit IL-12 transcription in response to interferon-gamma, lipopolysaccharide, and CD40 ligand and that Leishmania spp. lipophosphoglycan, phosphoglycans, and major surface protein are not necessary for inhibition. In addition, all the Leishmania spp. strains and life-cycle stages tested inhibited IL-12 promoter activity. Our data further reveal that autocrine-acting host factors play no role in the inhibitory response and that phagocytosis signaling is necessary for inhibition of IL-12.

Monoclonal antibodies (MAb) were prepared against the 40-kDa capsid protein of Cryptosporidium parvum virus (CPV) by immunizing mice with purified recombinant CPV40 protein. In immunoblotting analysis, MAbCPV40-1 bound to a 40-kDa protein in extracts of C. parvum oocysts. This 40-kDa protein was localized in the sporozoite cytoplasm by immunofluorescence (IFA) staining with MAbCPV40-1. In a dot-blot assay, MAbCPV40-1 was capable of detecting 10² non–bleach-treated and 10²–10³ bleach-treated C. parvum oocysts. MAbCPV40-1 was capable of detecting CPV40 antigen in both soluble and total C. parvum oocyst protein extracts, indicating a potential use for detecting this parasite in environmental samples.

The immune response of a host infected with Trypanosoma brucei is modulated by trypomastigotes. We examined the changes in cytokine production in T. brucei gambiense (Wellcome strain; WS) infected rats and the influence on production of interleukin (IL)-12 by macrophages. The blood concentration of interferon-γ, tumor necrosis factor–α, and IL-10 increased beginning the second day after infection. However, an increase in IL-12p40 was not observed until 4 days after infection. When spleen macrophages and Kupffer cells harvested from uninfected rats and HS-P cells (a rat macrophagelike cell line) were cocultured with WS, IL-12p40 production did not change. When HS-P cells were cultured with WS, transport of nuclear factor– κB into the nucleus increased. Levels of macrophage colony-stimulating factor (M-CSF) and granulocyte macrophage colony-stimulating factor mRNA in the spleens and livers of WS-infected rats were high in comparison with uninfected rats, suggesting that the WS promotes macrophage proliferation. The level of IL-12p40 mRNA in HS-P cells cocultured with WS increased in response to transfection with a small interfering RNA against M-CSF or addition of anti–M-CSF antibody. These results suggest that the WS inhibits IL-12p40 mRNA production by promoting production of macrophage colony-stimulating factor by macrophages.

Feeding polyamine-deficient chow (PDC) to rats decreases blood polyamines, increases the activity of ornithine decarboxylase as an index of polyamine production, and increases resistance to Trypanosoma brucei gambiense (Wellcome strain) (WS) infection. In this study, we investigated the influence on cytokine and nitric oxide (NO) production of feeding PDC to rats infected with WS. At 4 days postinfection with WS, serum concentration of interleukin (IL)-12, tumor necrosis factor-α, interferon-γ, IL-10, and NO increased in PDC-fed rats; however, IL-12 concentration in normal chow (NC)-fed rats did not increase. In spleen cells cocultured with WS, levels of IL-12 and inducible NO synthase (NOS) mRNA expression were higher in PDC-fed rats than in NC-fed rats. Proliferation of WS in coculture with spleen cells from PDC-fed rats was inhibited, but inhibition of WS proliferation was not observed when an NOS inhibitor was added into the culture media. Ornithine decarboxylase (ODC) activity increased in NC-fed rats after WS infection, but decreased in PDC-fed rats. These results show that feeding WS-infected rats PDC influences the production of cytokines such as IL-12 and the regulation of NO and polyamine production, and also leads to an increase in resistance against WS.

Physical or psychological stressors are known to have significant consequences for immune function and the outcome of disease in human and animal models. In mice, cold water stress (CWS) has been shown to delay control of acute infection and reactivation of latent infections. Increased levels of parasite-specific IgG and IgM antibodies are observed when CWS is applied in the chronic phase. The present study examined the effects of a physical stressor, CWS, on tachyzoites antigens of Toxoplasma gondii, with particular emphasis on a low molecular weight antigen, 5 kDa, which seems to be recognized by antibodies from mice subjected to CWS in the chronic phase. This antigen is not recognized by antibodies from infected mice not subjected to CWS. Sera obtained from stressed and infected (CWS INF) mice subjected to CWS during the chronic phase (CWS INF CWS) were used to harvest anti–5-kDa antibodies for immunolocalization studies. Tachyzoite lysate preparations were electrophoretically separated and transferred to nitrocellulose membranes. Strips of nitrocellulose containing tachyzoite antigens in the 4–10-kDa range were used to select for anti–5-kDa antibodies. Harvested anti–5-kDa localized this antigen on the surface of tachyzoites. This antigen was not present in bradyzoite preparations. Treatment with phosphatidylinositol-specific phospholipase C showed this antigen was not anchored to the cell membrane through glycosyl-phosphatidylinositol. Strong antibody responses in stressed animals during the chronic phase are associated with parasite reactivation. The 5-kDa antigen constitutes a unique immunogenic component of T. gondii, with significant diagnostic potential for identifying reactivation of latent infections.

mRNA and protein expression profiles for heat shock protein 70 (Hsp70) of Babesia gibsoni (BgHsp70) exposed to either high or low temperatures, were examined by quantitative real-time reverse transcription-polymerase chain reaction and Western blotting. In the present study, a commercially available anti-human Hsp70 antibody that could recognize recombinant BgHsp70 was used. BgHsp70 was detected in the parasites cultured under standard conditions at 37 C by Western blotting and immunostaining of thin smears of infected erythrocytes. These results suggested that BgHsp70 was expressed constitutively at the erythrocyte stage. BgHsp70 levels were elevated when the parasites were incubated at 42 C for 1 hr. In contrast, its mRNA amount was decreased and its protein amount was unchanged when the parasites were incubated at 32 C for 1 hr. Moreover, the level of parasitemia of B. gibsoni incubated at either 42 C or 32 C was almost the same as that at 37 C. These results indicated that the exposure of B. gibsoni to elevated temperatures might result in increased expression of BgHsp70 and that the exposure of the intraerythrocytic parasites to decreased temperatures might not induce the overexpression of BgHsp70.

Species of Perkinsus are responsible for high mortalities of bivalve molluscs world-wide. Techniques to accurately estimate parasites in tissues are required to improve understanding of perkinsosis. This study quantifies the number and tissue distribution of Perkinsus marinus in Crassostrea virginica by modern stereology and immunohistochemistry. Mean total number of trophozoites were (mean ± SE) 11.80 ± 3.91 million and 11.55 ± 3.88 million for the optical disector and optical fractionator methods, respectively. The mean empirical error between both stereological approaches was 3.8 ± 1.0%. Trophozoites were detected intracellularly in the following tissues: intestine (30.1%), Leydig tissue (21.3%), hemocytes (14.9%), digestive gland (11.4%), gills (6.1%), connective tissues (5.7%), gonads (4.1%), palps (2.2%), muscle (1.9%), mantle connective (0.8%), pericardium (0.7%), mantle epithelium (0.1%), and heart (0.1%). The remaining 0.6% were found extracellularly. Percentages of trophozoite stages were (mean ± SE): large, log-phase trophonts, i.e., signet rings, 97.0 ± 1.2%; meronts, 2.0 ± 0.9%; clusters of small, log-phase trophonts, i.e., merozoites, 1.0 ± 0.5%. Levels of infection in hemocytes and Leydig tissue were representative of total parasite intensity. These techniques are a powerful tool to follow parasite distribution and invasion, and to further explore mechanisms of Perkinsus spp. pathogenesis in bivalves.

The sequential developmental cycle of Haplosporidium montforti, a recently described species from farmed abalone Haliotis tuberculata (Gastropoda), was studied. Ornamented and operculated mature spores were electron dense. The nucleus of the uninucleated free cell divided successively, giving rise to multinucleate plasmodia, containing up to 100–120 nuclei. Later, the plasmodia developed into sporonts inside sporocysts with irregular contours. Each of their nuclei gave rise to uninucleate sporoblasts. At the next phase of development, a very irregular membranous group of cisternae began to differentiate in the cytoplasm of each sporoblast, surrounding each nucleus and the adjacent cytoplasm. Each sporoblast differentiated into a spore. This process was characterized by the appearance of dense blisters of amorphous material at the periphery that gradually formed the prespore wall and pre-operculum. Simultaneously, in the endosporoplasm, the spherulosome and several haplosporosomes were formed. During the final phase of the maturation process, the spores became gradually denser, and the endosporoplasmic structures were barely visible.

Bottlenose dolphins (Tursiops truncatus) captured in the estuarine waters off the coasts of South Carolina and Florida were examined for the presence of Microsporidia, Cryptosporidium sp., and Giardia sp. DNA extracted from feces or rectal swabs was amplified by polymerase chain reaction using parasite-specific small subunit ribosomal RNA gene primers. All positive specimens were subjected to gene sequence analysis. Of 83 dolphins, 17 were positive for Microsporidia. None was positive for Cryptosporidium or Giardia. Gene sequence data for each of the positive specimens were compared with data in GenBank. Fourteen specimens were found similar to, but not identical to, the microsporidian species Kabatana takedai, Tetramicra brevifilum, and Microgemma tinca, reported from fish, and possibly represent parasites of fish eaten by dolphins. Gene sequence data from 3 other specimens had ∼87% similarity to Enterocytozoon bieneusi, a species known primarily to infect humans and a variety of terrestrial mammals, including livestock, companion animals, and wildlife. It is not clear if these specimens represent a species from a terrestrial source or a closely related species unique to dolphins. There were neither clinical signs nor age- or gender-related patterns apparent with the presence of these organisms.

The present study provides the first record of Trypanosoma chattoni Mathis and Leger, 1911, in a new host, Leptodactylus fuscus Schneider, 1799 (Anura, Leptodactylidae), and the occurrence of Trypanosoma rotatorium–like species in Leptodactylus chaquensis Cei, 1950. The anurans were captured in the State of Mato Grosso, Brazil. Blood samples were obtained by cardiac puncture, and blood smears were examined for the presence of hemoparasites. The Trypanosoma rotatorium–like species in this study refers to a short-bodied trypomastigote that has a conspicuous undulating membrane but lacks a free flagellum; T. chattoni refers to a monomorphic parasite that has a rounded body, a kinetoplast adjacent to the nucleus, and a short flagellum.

A cross-sectional study of 27 cohorts of Tribolium confusum aged 2–78 wk was done to examine effects of host age on exposure to eggs of Hymenolepis diminuta under standardized conditions. Pre-exposure, fasting, and postexposure mortality were low, sex ratio was equal, and fecundity of hosts was high during the first 30 wk, followed by increasing mortality and male bias of the sex ratio, and declining fecundity, in older beetles. These changes in the host were not associated with pronounced changes in infection results. Prevalence of infection was higher in females than males, but was unaffected by age in both sexes. Intensity of infection was similar between sexes in beetles up to 30 wk old, and thereafter declined in females, but not in males. Age-related changes in hosts were gradual, but unexpected levels of short-term variation in infection results suggest that some undetermined proximate factors may override general host age effects on the infection process.

This study presents the first characterization of a WD40 repeat-containing myosin identified in the apicomplexan parasite Gregarina polymorpha. This 222.7 kDa myosin, GpMyoF, contains a canonical myosin motor domain, a neck domain with 6 IQ motifs, a tail domain containing short regions of predicted coiled-coil structure, and, most notably, multiple WD40 repeats at the C-terminus. In other proteins such repeats assemble into a β-propeller structure implicated in mediating protein-protein interactions. Confocal microscopy suggests that GpMyoF is localized to the annular myonemes that gird the parasite cortex. Extraction studies indicate that this myosin shows an unusually tight association with the cytoskeletal fraction and can be solubilized only by treatment with high pH (11.5) or the anionic detergent sarkosyl. This novel myosin and its homologs, which have been identified in several related genera, appear to be unique to the Apicomplexa and represent the only myosins known to contain the WD40 domain. The function of this myosin in G. polymorpha or any of the other apicomplexan parasites remains uncertain.

Microsporidia are obligate intracellular, eukaryotic parasites that are known to infect a variety of invertebrate and vertebrate species and have been reported to include a broad range of host specificities for various cell types. Although it is clear that some species of microsporidia have the ability to disseminate, causing multiorgan infections, it is not understood how dissemination occurs. One hypothesis suggests that mononuclear phagocytes engulf the pathogen and migrate to various organs while the parasite persists and proliferates. This implies that microsporidia have developed methods by which to escape intracellular degradation and can, instead, use the host as a source of nourishment and a vehicle for dissemination. In our study, we investigated the infection kinetics of 2 Encephalitozoon spp. known to cause disseminated disease in humans. Using fluorescence and scanning electron microscopy, it was determined that spore adherence to the host was rapid (3–6 hr), as was the uptake and organization of internal parasitophorous vacuoles (24 hr). Furthermore, replication was shown to occur within macrophages at 72 hr, as measured by the bromodeoxyuridine proliferation assay, and the production of mature spores occurred in host cells at 120 hr. Parasitic replication could be reduced by pretreatment of macrophages with interferon-gamma and bacterial lipopolysaccharide.

A new species of Allocreadium, Allocreadium danjiangensis n. sp., is described from the intestine of several species of freshwater fish, including Abbottina rivularis (Basilewsky, 1855), Sarcocheilichthys nigripinnis nigripinns (Günther, 1873), Gnathopogon argentatus (Sauvage et Dabry 1874), Opsariichthys uncirostris bidens (Gunther, 1873), and Erythroculter mongolicus mongolicus (Basilewsky, 1855) (Cyprinidae) from the Danjiangkou Reservoir in central China. The main morphological characters of the new species are as follows: vitelline follicles numerous, extending from the level of acetabulum to posterior extremity, distributed over both sides around the ceca; cirrus sac relatively large, developed, lying obliquely anterior to the acetabulum, extending from the level of the intestinal bifurcation to the central level of acetabulum, and overlapping left or right cecal; and ovary much smaller than testes, generally close to or even overlapping the anterior border of anterior testis. Observation by scanning electron microscopy shows only 2 kinds of tegumental formations, i.e., papillae and tubercles, instead of 3 types of tegumental formations, i.e., papillae, bosses, and minute sensor receptors observed on other species of the Allocreadiidae. The tegumental striations of the present species vary on the different parts of the body. In addition, a new structure, identified as the “groove” with a tonguelike tubercle, was observed on the inner wall of acetabulum.

Adults of 2 new species in 2 orders of acanthocephalans obtained from the intestines of terrestrial amphibians and reptiles collected between 1998 and 2004 in Vietnam are described here. Pseudoacanthocephalus nguyenthileae n. sp. (Palaeacnthocephala: Echinorhynchidae) was collected from 5 species of terrestrial amphibians: (1) the common Sunda toad Bufo melanostictus Schneider (Bufonidae); (2) Paa verucospinosa (Bourret); (3) Gunther's Amoy frog Rana guentheri Boulenger; (4) Taipei frog R. taipehensis Denburgh (Ranidae), and (5) the Burmese whipping frog Polypedates mutus (Smith) (Racophoridae); as well as from the Chinese cobra Naja atra Cantor (Reptilia: Elapidae) and house gecko Hemidactylus frenatus Dumeril and Bibron (Reptilia: Gekkonidae). Sphaerechinorhynchus maximesospinus n. sp. (Plagiorhynchidae: Sphaerechinorhynchinae) was isolated from a king cobra Ophiophagus hannah (cantor) (Reptilia: Elapidae). Cystacanths of Porrorchis houdemeri (Joyeux and Baer, 1935) Schmidt and Kuntz, 1967 (Plagiorhynchidae: Porrorchinae) obtained from the mesenteries of banded krait Bungarus fasciatus (Schneider) (Reptilia: Elapidae), a paratenic host, are reported for the first time. Keys to the species of Pseudoacanthocephalus and Sphaerechinorhynchus are included. Characteristic features distinguishing the new species from related taxa include: P. nguyenthileae has 15–19 (usually 16–18) proboscis hook rows, each with 5–6 hooks that progressively increase in length and size posteriorly. The largest, intermediate, and smallest proboscis hooks of S. maximesospinus are the middle, anterior, and posterior hooks, respectively; the proboscis and neck are enclosed in a membrane. Morphometric characteristics of P. nguyenthileae show host-related variability.

Parathelandros allisoni n. sp. from the intestines of the frog Nyctimystes trachydermis (Hylidae) from Central Province, Papua New Guinea is described and illustrated. It is the 12th species assigned to the genus and the ninth species of Parathelandros to be reported from the Australo-Papuan region. Parathelandros allisoni n. sp. is most similar to Parathelandros australiensis in that the vulva occurs near the esophageal bulb, but differs in that males lack the spherical postcloacal lobe characteristic of P. australiensis. The description of P. andersoni is amended to include operculate eggs and rosette papillae on the tail of the male.

Adults of 2 new species and 2 new genera of acanthocephalans in class Archiacanthocephala, collected between 1998 and 2004 in Vietnam from the intestines of mammals, are described, i.e., Cucullanorhynchus constrictruncatus n. gen., n. sp. (Oligacanthorhynchidae) from a leopard Panthera pardus (Linnaeus) (Mammalia: Felidae) and Paraprosthenorchis ornatus n. gen. n. sp. (Oligacanthorhynchidae) from the Chinese pangolin Manis pentadactyla (Linnaeus) (Mammalia: Manidae). Adult Sphaerechinorhynchus macropisthospinus Amin, Wongsawad, Marayong, Saehoong, Suwattanacoupt, and Sey, 1998 (Plagiorhynchidae) are described for the first time from 2 females collected from a tiger Panthera tigris (Linnaeus) (Mammalia: Felidae) and from 1 male from a water monitor Varanus salvator Laurenti (Reptilia: Varanidae). Characteristic features distinguishing the new species or genera from related taxa are as follows. The trunk of C. constrictruncatus has an anterior hood in both sexes and a posterior constriction in females. The anterior trunk of P. ornatus has many small festoons and proboscis hooks are inserted in elevated papillae separated by beady, near hexagonal, ornate grids.

Sandonella sandoni (Lynsdale, 1960) is the type and only species of the Sandonellinae, a cestode subfamily of unclear phylogenetic position. It is redescribed here on the basis of a re-examination of its syntypes, voucher specimens from museum collections, and freshly collected material from the intestine of Heterotis niloticus (Osteoglossiformes: Arapaimidae) from Benin, Nigeria, Senegal, and the Sudan. The species possesses several unique morphological characters, such as (1) a vitellarium formed by 2 compact, but deeply lobulated, postovarian masses near the posterior margin of proglottids; (2) a scolex with a highly modified apical structure formed by 4 muscular retractile lappets; (3) a well-developed circular musculature, which is external to the inner longitudinal muscles; (4) a dilated, vesicle-like proximal part of the external sperm duct; (5) the unique morphology of the uterus and its development, which represents an intermediate form between the 2 basic types recognized in the Proteocephalidea; (6) the growth of eggs during their development within the uterus; and (7) the complex proglottization with intermingled smaller and larger (wider) proglottids. The morphology of S. sandoni, including the form and distribution of microtriches, was studied by scanning electron microscopy for the first time, and the lectotype and paralectotypes of S. sandoni are designated. Sequences of the 28S rRNA gene of 4 specimens (2 from the Sudan and 2 from Senegal) were identical, which confirms conspecificity of geographically distant samples. Sandonella sandoni sequences have also shown that it actually belongs among the Proteocephalidea, being a sister taxon of a relatively derived clade of Palaearctic proteocephalideans, containing Glanitaenia osculata and Paraproteocephalus parasiluri from catfish and Palaearctic species of the Proteocephalus aggregate.

Members of the Polymorphidae (Acanthocephala) are distributed worldwide as endoparasites of marine mammals, fish-eating birds, and waterfowl. The family contains 10 genera, with approximately 127 species. Polymorphids are characterized by having a spinose trunk with a bulbous proboscis, double-walled proboscis receptacle, long lemnisci, and 4 tubular cement glands. The taxonomic position of several genera within Polymorphidae has been controversial when considering morphological and ecological characters. The mitochondrial coding gene cytochrome-c oxidase representing species of 5 genera of polymorphids (Corynosoma, Lühe, 1904, Hexaglandula Petrochenko, 1950, Southwellina Witenberg 1932, Polymorphus Luhë, 1911, and Profilicollis Meyer, 1931) were sequenced to determine the sister-group relationships among 2 particular genera, i.e., Hexaglandula, and Profilicollis. Maximum likelihood and Bayesian analyses showed that Polymorphidae is a monophyletic assemblage, and that 3 major clades are present. Our results provide support for the idea that Hexaglandula represents an independent lineage, whereas, in the case of Profilicollis, there is no conclusive evidence that they are not members of Polymorphus. The analyses also confirm that Polymorphus is paraphyletic, suggesting that the genus represents a complex of species that should be re-examined and reclassified using morphological, ecological, and molecular data. Our observations suggest that decapods (intermediate hosts for the 2 genera under study) were independently colonized at least twice during the evolutionary history of the group.

A new species of Myxosporea, Myxobolus neurotropus n. sp., is described from the brain and spinal cord of rainbow trout (Oncorhynchus mykiss) from Duncan Creek, Owyhee County, Idaho. Spores are oval, have 2 pyriform polar capsules, and possess a thick spore wall (sutural rim) with a short intracapsular offshoot. The mean spore dimensions are length 11.8 μm, width 10.8 μm, and thickness 8.8 μm. This myxozoan is compared to other described Myxobolus species found in cranial tissues of salmonids in terms of spore morphology and phylogenetic analysis. Because it is found in brain and spinal cord, it is encountered while performing screening tests for Myxobolus cerebralis, the causative agent of salmonid whirling disease. Where chronic inflammation and granulomatous lesions are associated with M. cerebralis, histological examination shows no host response to M. neurotropus n. sp. A diagnostic polymerase chain reaction (PCR) test is included as an aid in properly identifying the species.

A new species of dicyemid mesozoan is described from Sepiella japonica Sasaki, 1929, collected from the eastern area of Inland Sea, Osaka Bay, and Kii Strait in Japan. Dicyema sepiellae n. sp. is a large species that reaches about 4,000 μm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 24–29 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells, but to the base of propolar cells in vermiform embryos. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid. This is the first description of a dicyemid in any species of Sepiella.

Abomasal nematodes (Ostertagiine: Trichostrongyloidea) representing a previously unrecognized genus and species are reported in African buffalo (Syncerus caffer caffer) from Kenya, Uganda, and South Africa. Africanastrongylus buceros gen. nov. et sp. nov. is characterized by a symmetrical tapering synlophe in the cervical region and a maximum of 60 ridges in males and females. Bursal structure is 2–2–1, with subequal Rays 4/5, massive Rays 8, and Rays 9/10, and a massive dorsal lobe that is reduced in length, laterally and dorsally inflated, and positioned ventral to externodorsal rays. Spicules are tripartite, and the gubernaculum is broadly alate in the anterior. A proconus is present. Among ostertagiines with a 2–2–1 bursa (Cervicaprastrongylus, Hyostrongylus, Mazamastrongylus, Sarwaria, Spiculopteragia, and Teladorsagia) specimens of Africanastrongylus are differentiated from respective genera based on the structure of the cervical synlophe, patterns of dorsal, externodorsal, lateral, and ventral rays, and configuration of the genital cone, gubernaculum, and spicules. Among 13 genera of the Ostertagiinae in the global fauna, 3 are entirely limited in distribution to Africa, including Africanastrongylus, Longistrongylus, and Pseudomarshallagia. Another 5 genera including Cervicaprastrongylus, Hyostrongylus, Marshallagia, Ostertagia, and Teladorsagia are represented as mosaics, with diversity centered in Eurasia or the Holarctic. Genera not represented in the African fauna include Camelostrongylus among Caprinae and some Antelopinae from Eurasia, Mazamastrongylus and Spiculopteragia in Cervidae from the Holarctic and Eurasia, respectively, Orloffia in Cervidae and Bovidae from the Holarctic, and Sarwaria among Tragulidae and Bovinae in southern Asia. The diverse nature of the ostertagiine fauna, with a disproportionate number of endemic genera relative to other regions of the northern hemisphere, may reflect the timing of episodic expansion events for artiodactyls into Africa from Eurasia during the Tertiary and Quaternary.

In this study, 107 fecal specimens were collected from 40 sampling sites in Taiwan livestock and avian farms to test for Cryptosporidium spp. oocysts. Ten of 107 samples analyzed by enzyme-linked immunosorbent assay showed the presence of Cryptosporidium spp., among which 6 samples were simultaneously confirmed by immunofluorescence assay and polymerase chain reaction. Nucleic acid sequencing of the 18S rRNA gene identified 3 clusters of Cryptosporidium spp. Three Cryptosporidium parvum isolates were from cattle and sheep feces. One Cryptosporidium andersoni isolate was detected from pig feces. The other 2 novel Cryptosporidium genotypes were not similar to any known Cryptosporidium spp. according to the DNA sequences of the 18S rRNA gene.

A new species of Crassicutis Manter, 1936 is described from the Sinaloan cichlid Cichlasoma beani (Jordan) (Osteichthyes: Cichlidae) in the upper Río Santiago basin. Crassicutis choudhuryi n. sp. differs from most of the other nominal species by having testes located in a symmetrical position. The only other species of the genus that includes some specimens exhibiting this trait is Crassicutis intermedius (Szidat 1954), a species found in 5 species of siluriforms and 1 species of characiform in South America. However, this species differs from Cr. choudhuryi n. sp. by having testes almost half of the size, and vitelline follicles extending anteriorly to the region between the acetabulum and the intestinal bifurcation. The new species is morphologically very similar to Crassicutis cichlasomae Manter, 1936, but clearly differs from this species because of the constantly symmetrical position of the testes. Additionally, Cr. choudhuryi n. sp. is found in the Santiago River basin on the Pacific slope of Mexico, parasitizing specifically the endemic Ci. beani that does not co-occur with any other cichlid. Cr. cichlasomae exhibits more hosts (about 25 species of cichlids only in Mexico) and a wider distribution range that extends from northeastern Mexico southward to Central America, Cuba, and Brazil. To corroborate that our specimens were not conspecific with Cr. cichlasomae, sequences of the internal transcribed spacer (ITS1) and the 28S ribosomal RNA genes of individuals from several populations (recently collected in southeastern Mexico) were obtained and compared to the species described herein. Sequence divergence (1.3% for the 28S and 4.0% for the ITS1) gives further support to the erection of a new species.

Oswaldocruzia belenensis n. sp. (Strongylida: Molineidae) from the small intestine of Chaunus marinus (L.) from Belém, Pará State, Brazil is described and illustrated by light and scanning electron microscopy. Oswaldocruzia belenensis n. sp. is a neotropical species of this genus, harboring caudal bursa Type II, spicules divided in 3 branches, i.e., a blade, shoe, and fork. The blade is divided in 4 points, of which at least 2 are bifurcated. Cervical alae are absent; there is a simple cephalic vesicle and synlophe with low ridges perpendicular to the body without chitinous supports. The most closely related species are O. bonsi and O. lescurei. Oswaldocruzia belenensis n. sp. differs from O. lescurei and O. bonsi by the number and location of cephalic papillae, rays 2–3 and 5–6 running parallel and slightly separated, ray 6 not overlapping ray 8, and body structure morphometry. Oswaldocruzia belenensis n sp. also differs from O. lescurei by the discontinuity of the longitudinal ridges, the number of subdivisions of the blade, and the absence of extra processes at the bifurcation level of the fork of the spicules. The new species differs from O. bonsi by male and female body dimensions, the symmetry of the caudal bursa, dimension and subdivisions of the spicules without extra processes of the fork, 2 extra processes at the distal division of the blade, and location of ray 7 at the anterior margin of the cloacal aperture. Oswaldocruzia belenensis n. sp. represents the 82nd species assigned to the genus.

The myxozoan genus Davisia Laird 1953 is preoccupied by Davisia Del Guercio 1909 (Insecta: Hemiptera). Here, Myxodavisia nomen novum is proposed to replace the preoccupied name, and a new species is described. Myxodavisia sebastisca n. sp. was found in the urinary bladder of Sebastiscus marmoratus, collected from coastal waters off Xiamen in the East China Sea. The parasite is characterized by a disporous trophozoite; spherical to subspherical spore, 13.1 ± 0.3 (12.7–13.6) by 12.3 ± 0.9 (10.9–13.5) μm in size; curved sutural line; 2 shell valves each with a long lateral appendage 119.4–335.2 μm in length; and 2 spherical or subspherical polar capsules, equal in size, 4.6 ± 0.6 (3.2–4.6) μm in diameter. Traditionally, Myxodavisia is distinguished from Ceratomyxa, Sinuolinea, and Sphaerospora spp. by having spores that possess a distinct central chamber and lateral appendages. A review of the literature reveals that the presence or absence of a clear septum between these spore components is open to interpretation. Indeed, in immature spores of M. sebastisca n. sp., there was an indication of a demarcated appendage, but in some mature spores, no clear separation was apparent. Our findings suggest that future revision of this genus is warranted, particularly once DNA sequence data become available.

This paper evaluates the effects of certain herbicides on Leishmania spp., their mechanism of action, and the evolutionary origin of the relevant susceptible leishmanial targets. We demonstrated that a relatively nontoxic herbicide, fenarimol, successfully interferes with a leishmanial target, which is probably a relic of an ancient ancestor. Fenarimol impairs the function of leishmanial 14α-sterol demethylase, a key enzyme in the sterol biosynthetic pathway. Therefore, fenarimol or its derivatives may be candidates for development of anti-leishmanial drugs. Of the herbicides that have the capability to act as potential inhibitors of the metabolism of Leishmania spp., fenarimol was found as the most active substance against both promastigotes and amastigotes in culture. In addition, it ameliorated lesions caused by Leishmania major in mice. Light microscopy demonstrated rounding of the parasite shape. Increase of osmophilic vacuoles and autophagosomal structures were observed by transmission electron microscopy. Biochemical studies demonstrated that fenarimol inhibited sterol biosynthesis. Docking of fenarimol to the modeled catalytic binding site of 14α-lanosterol demethylase of L. major showed a geometrical fit. Fenarimol is stabilized via hydrophobic interactions with the residues that surround it and interactions with the heme ring. These results provide support to the hypothesis that fenarimol inhibits leishmanial sterol biosynthesis. Overall, the findings suggest an additional source of substances for development of anti-leishmanial drugs.

Saimiri boliviensis monkeys were infected via sporozoites with the Salvador I strain of Plasmodium vivax that had been stored frozen for periods ranging from 12 to 5,312 days. Prepatent periods ranged from 16 to 53 days.

In total, 9 endohelminth species were found to parasitize 7 fish species (2 cyprinids, 4 goodeids, and 1 poeciilid) from La Mintzita Reservoir, Michoacán, in central Mexico; 5 were larvae, including 3 allogenic species (Clinostomum complanatum, Tylodelphys sp., Posthodiplostomum minimum) and 2 autogenic species (Serpinema trispinosum, Spiroxys sp.). Four were enteric autogenic adults, i.e., Margotrema bravoae, Bothriocephalus acheilognathi, Proteocephalus longicollis, and Rhabdochona lichtenfelsi. The metacercariae of P. minimum reached the highest levels of prevalence and mean abundance among host species. Our results confirm the depauperate nature of the helminth communities of freshwater fishes from central Mexico. On the basis of this data set, we estimated the total endohelminth species richness for each component community by using 7 nonparametric estimators whose performance was evaluated with the unscaled measures of bias, precision, and accuracy. We found that Chao1 and Bootstrap are the most precise and least biased methods for the 7 component communities; however, species richness was consistently underestimated. The underestimation was an unavoidable consequence of the patchy distribution of helminth species among different component communities, particularly at the small sample size used in our study.

To estimate isotopic changes caused by trematode parasites within a host, we investigated changes in the carbon and nitrogen isotope ratios of the freshwater snail Lymnaea stagnalis infected by trematode larvae. We measured carbon and nitrogen stable isotopes within the foot, gonad, and hepatopancreas of both infected and uninfected snails. There was no significant difference in the δ¹³C and δ¹⁵N values of foot and gonad between infected and uninfected snails; thus, trematode parasite infections may not cause changes in snail diets. However, in the hepatopancreas, δ¹⁵N values were significantly higher in infected than in uninfected snails. The ¹⁵N enrichment in the hepatopancreas of infected snails is caused by the higher ¹⁵N ratio in parasite tissues. Using an isotope-mixing model, we roughly estimated that the parasites in the hepatopancreas represented from 0.8 to 3.4% of the total snail biomass, including the shell.

Ticks have never been reported in archaeological analyses. Here, we present the discovery of a tick from a coprolite excavated from Antelope Cave in extreme northwest Arizona. Dietary analysis indicates that the coprolite has a human origin. This archaeological occupation is associated with the Ancestral Pueblo culture (Anasazi). This discovery supports previous hypotheses that ticks were a potential source of disease and that ectoparasites were eaten by ancient people.

Cryptosporidium parvum (HNJ-1 strain, genotype 2) merozoites were released from oocysts directly during an incubation and excystation procedure without bleach treatment. They were polymorphic, mostly spindle-shaped; others were bean shaped, actively motile, and underwent division. Merozoites survived for short time–period in an in vitro culture system, but could not be established in a subsequent cultivation effort in RPMI medium.

Parasitological analysis of 5 sediment samples from San Bernardino, California latrine deposits spanning the time period from about 1880 to the 1930s are presented. Two sediment samples are from a latrine used by European-Americans. Three sediment samples are from latrines used by Chinese-Americans on the property of Wong Nim, an important member of the Chinese community. Two of the Chinese latrines were positive for human parasites. The human parasites encountered include the human whipworm (Trichuris trichiura), the giant intestinal roundworm (Ascaris lubricoides, c.f.), and the Chinese liver fluke (Clonorchis sinensis). Evidence of the liver fluke is especially important. This parasite cannot complete its life cycle outside of its endemic range in Asia because suitable intermediate hosts are not present in the American continents. Its presence signals that at least some of the Chinese-Americans who used the latrines were immigrants who were infected in Asia and then sustained infections while in the Americas.

Neospora caninum is one of most important causes of abortion in cattle worldwide, and dogs are an important risk factor for N. caninum infection in cattle. Antibodies to N. caninum were determined in 184 (126 rural, 58 urban) dogs from the Punjab State, India, using commercial monoclonal antibody-based competitive ELISA and found in 16.8% of the animals. The prevalence of N. caninum antibodies was significantly higher in rural dogs (21.4%, 27 of 126) than city dogs (6.9%, 4 of 58). To our knowledge this is the first report of N. caninum infection in canines from India.