The acanthocephalan parasite Acanthocephalus dirus develops from the egg to the cystacanth stage inside the freshwater isopod Caecidotea intermedius. We have shown previously that cystacanth-infected male C. intermedius are less likely to initiate mating attempts with females than uninfected males in competitive situations. Here, we used a field-based experiment to examine whether cystacanth-infected males were also less likely to initiate mating attempts with females in noncompetitive situations. We found that infected males were less responsive to females than uninfected males, and we propose that the cystacanth-related change in male mating behavior is mediated by a change in the mating response of males to females rather than male–male competition. We then examined whether cystacanth-related changes in reproductive function, i.e., sperm content and fertilization ability, could explain this variation in male mating behavior. We found that cystacanth-infected males contained both developing and mature sperm and fertilized as many eggs as uninfected males. Thus, we propose that changes in reproductive function are unlikely to explain cystacanth-related variation in male mating behavior in C. intermedius.

We examined the effect of isopod size and age on the success of an acanthocephalan infection and on the effects of that infection on the growth and survival of the isopods. Groups of isopods (Asellus aquaticus) belonging to 4 size classes (juveniles, maturing adults, young adults, and older adults) were exposed to infective acanthors of Acanthocephalus lucii. At the end of the experiment, survival of the isopods, lengths of male and female isopods, and numbers of different developmental stages of A. lucii larvae in infected isopods were assessed. Acanthocephalus lucii prevalence was significantly lower in juvenile isopods than in adults. Intensity of infection increased with the size of isopods at exposure, and cystacanth intensity correlated positively with isopod size at the end of the experiment. Exposed juveniles and maturing adults survived significantly better than unexposed individuals, but the opposite was true of the 2 largest size classes. At the end of experiment, exposed isopods, and, especially, cystacanth-infected isopods, were significantly larger than unexposed isopods in every size class. We suggest that isopod size not only affects the success of A. lucii infection but also affects the ability of A. lucii to affect the survival (and perhaps the growth) of the isopod hosts.

Spermiogenesis and the spermatozoon of Crepidostomum metoecus, an intestinal parasite of brown trout Salmo trutta, were studied by transmission electron microscopy. Spermiogenesis begins with the formation of a differentiation zone in front of 2 centrioles associated by an intercentriolar body. Each centriole is linked to a striated rootlet, and gives rise to a flagellum. The rotation of flagella is greater than 90°; their fusion with the median cytoplasmic extension is proximodistal and asynchronous. The spermatozoon is formed after constriction of arched membranes. The spermatozoon possesses 2 axonemes of the 9 “1” pattern, a nucleus, mitochondria, and glycogen. A major feature is the presence, in the anterior part, of external ornamentation and a lateral expansion associated with spinelike bodies. Another attribute is the presence of 2 mitochondria rather than just 1, as in most of the digenean spermatozoa. To our knowledge, this study is the first undertaken with a species of the Allocreadiidae.

Spermiogenesis and ultrastructure of spermatozoon of Nicolla wisniewskii (Digenea, Opecoelidae), an intestinal parasite of Salmo trutta, were studied by electron microscopy. Spermiogenesis follows the general pattern found in the Digenea. It begins with the formation of a differentiation zone, including striated rootlets associated with 2 centrioles and an intercentriolar body. The flagella undergo a rotation of greater than 90°. Then, their fusion with the median cytoplasmic process is proximodistal and asynchronous. A peculiarity was observed before the fusion of flagella, i.e., the attachment zones joined as 2 pairs by an electron-dense bridge. The mature spermatozoon is characterized by 2 axonemes, cortical microtubules, a nucleus, 2 mitochondria, external ornamentation, and spinelike bodies. At the posterior end of flagella, the spermatozoon is also characterized by the presence of a central element of the axoneme and without the 9 microtubule doublets. These results were compared with those of the other digeneans and, in particular, with other species of Opecoelidae. It appears that the number of cortical microtubules and their localization in the spermatozoon may be an interesting feature of their phylogeny.

In Gregarina niphandrodes, an apicomplexan parasite, the sexual stage of its life cycle begins with the association of 2 gamonts. Here, we describe the ultrastructure of the syzygy junction and the nucleus during the transition from unassociated trophozoites to paired gamonts to gamonts in syzygy. Throughout this process, the folds within the syzygy junction undergo changes that correspond to changes of the epicytic folds. The nucleus goes through dramatic changes from multiple spheres of condensed chromatin in unassociated trophozoites, to mostly uncondensed chromatin in paired gamonts, to a large single sphere of condensed chromatin encasing many smaller spheres in gamonts in syzygy. These differing nuclear ultrastructures reflect the dramatic cellular and transcriptional changes associated with life cycle transitions and are indicative of the numerous cell divisions that follow.

Helminth communities in sympatric black turnstones (Arenaria melanocephala), ruddy turnstones (Arenaria interpres), and dunlin (Calidris alpina) were examined over 4 summers in Bristol Bay, Alaska. The compound community, made up of component communities of all 3 species of hosts for 4 summer seasons (n = 164), consisted of 43 helminth species, with cestodes, especially Anomotaenia clavigera, accounting for 47% of the helminth species and 95% of the abundance. The black turnstone had significantly higher species richness and abundance than either the ruddy turnstone or dunlin. The congeneric black and ruddy turnstone component communities were the most similar, and the dunlin's was the least similar. New helminth species continued to be acquired in all 3 host species during years 2 to 4. There was no significant difference for abundance among sample years for each of the 3 species of host. The 3 component communities all included a predictable suite of helminths with 1 dominant species and 4 to 5 associates, a large number of less-predictable species, and a greater prevalence and abundance of cestode species. Consistencies over time included high diversity, low evenness, low species richness (<5), and continued recruitment of small numbers of helminth species with low prevalence and abundance. There was minimal circulation of helminth species between the dunlin and the 2 turnstone species, indicating a considerable degree of specialization, particularly among species of cestodes.

Previous surveys of wild ungulates indicate that the liver fluke, Dicrocoelium dendriticum, was rare in the Cypress Hills area of southeastern Alberta. However, 41 of 59 wapiti (Cervus elaphus) sampled during the 2003 and 2004 hunting seasons from this region were infected, with 7 hosts containing >1,000 worms. Prevalence and mean intensity were similarly high in sympatric beef cattle and mule deer. Worm abundance in wapiti was age related, with calves containing significantly higher numbers of worms (mean ± SD abundance = 825 ± 1,098) than adults (107 ± 259). This pattern with host age was not evident in beef cattle, although the smaller sample sizes may be a contributing factor. These results indicate that D. dendriticum is now well established in Cypress Hills Park, circulating between at least 3 species of sympatric ungulates, including beef cattle.

Galápagos penguins (Spheniscus mendiculus) and flightless cormorants (Phalacrocorax harrisi) live in small, isolated populations on the westernmost islands of Isabela and Fernandina in the Galápagos Islands, Ecuador. Between August 2003 and February 2005, 4 field trips, 2 in the cool, dry season (August 2003 and August 2004) and 2 in the hot, rainy season (March 2004 and February 2005), were undertaken; 298 Galápagos penguins and 380 cormorants were sampled for prevalence and intensity of hemoparasites. Microfilariae were found in both the penguins and the cormorants. Blood smears were negative for the presence of other species of hemoparasites. Overall prevalence of microfilariae across seasons was 42.0% in cormorants and 13.8% in the penguins. Intensity of infection was generally low (mean = 3.2–31.7 in 25 fields across seasons and species) with the exception of a few individuals with markedly high intensities of parasites (>300 in 25 fields in 1 cormorant). Prevalence of microfilariae increased significantly over the 4 sampling periods for cormorants, but not for penguins. Prevalences were significantly higher in cormorants than in penguins for 3 of the 4 collecting trips. Male penguins had higher prevalences than females; however, there were no gender differences in cormorants. No relation was detected between body mass and either presence or intensity of parasitism. Morphological characteristics of the microfilariae are also described and specimens from each host species were similar in all characters measured. DNA sequence data from the mitochondrial cytochrome c oxidase subunit I gene were consistent with the morphological evidence and together demonstrate that the penguins and cormorants are likely to be infected with the same species of microfilariae.

Male vertebrates are believed to be disproportionately vulnerable to parasites, but empirical support for this contention is mixed. We tested the hypothesis of higher levels of parasitism in males with the use of counts of gastrointestinal helminths in 5 sympatric mammalian carnivores (American badgers, coyotes, red foxes, raccoons, striped skunks) from central Saskatchewan. Parasite burdens for females and males of each host species were compared with the use of prevalence (percentage of hosts infected), intensity (parasites per infected host), and overdispersion (proportion of heavily infected hosts that were male). Of 30 comparisons (13 each for prevalence and intensity, 4 for overdispersion), male bias was detected 8 times (27%), whereas female bias was detected only once (3%), adding some support to the notion that male mammals are more susceptible to parasitism. However, most of the statistical comparisons we undertook revealed no sexual bias (n = 21, 70%), suggesting that differential patterns of infection are not ubiquitous in mammals. Moreover, when detected, the magnitude and direction of bias varied among host species, helminth species, and metrics of infection. We conclude that sympatric and ecologically similar mammal species will not always share the tendency for males to be more susceptible to parasitism, and that studies incorporating multiple parasites and metrics of infection are more likely to detect sex bias.

We examined the role of lizards in the ecology of Lyme disease in New York and Maryland. We collected data on vector tick infestations, measured lizard “realized” reservoir competence for the Lyme disease spirochete Borrelia burgdorferi, and estimated lizard population density. These data were incorporated into a model that predicts a host's ability to influence the prevalence of B. burgdorferi in the tick population, a primary risk factor in the epidemiology of Lyme disease. Published data on other northeastern hosts were included in the model to provide a reference for interpreting the importance of lizards as hosts. The model results indicate that 5-lined skinks (Eumeces fasciatus) are dilution hosts, capable of reducing infection prevalence in the tick population by 10.7–51.5 percentage points, whereas eastern fence lizards (Sceloporus undulatus) are not dilution hosts in the areas studied. Owing to moderate burdens of larval ticks, relatively high population densities, and reservoir incompetence, E. fasciatus may play an important role in the ecology of Lyme disease by reducing vector infection prevalence and associated human risk of infection.

Ectoparasites of an urban population of big brown bats (Eptesicus fuscus) in Fort Collins, Colorado, were investigated during summers 2002, 2003, and 2004. Eleven species of ectoparasites were found (the macronyssid mite Steatonyssus occidentalis, the wing mite Spinturnix bakeri, the myobiid mites Acanthophthirius caudata and Pteracarus aculeus, the chirodiscid mite Alabidocarpus eptesicus, the demodicid mite Demodex sp., the chigger Leptotrombidium myotis, the soft tick Carios kelleyi, the batfly Basilia forcipata, the batbug Cimex pilosellus, and the flea Myodopsylla borealis). Five species were analyzed by prevalence and intensity (C. pilosellus, M. borealis, L. myotis, S. bakeri, and S. occidentalis) based on 2,161 counts of 1,702 marked individual bats over the 3 summer study periods. We investigated 4 factors potentially influencing prevalence and intensity: age class of the host, reproductive status of adult female hosts, roosts in which the hosts were found, and abiotic conditions during the year sampled. The macronyssid mite, S. occidentalis, was the most prevalent and abundant ectoparasite. Adult big brown bats had more ectoparasites than volant juveniles for most of the species analyzed. In a sample of known age bats at 1 large colony, bats of 4 yr of age or greater had higher ectoparasite loads of S. occidentalis and S. bakeri when compared with younger bats. Lactating female bats had the highest prevalence and intensities of most ectoparasites. Annual differences in ectoparasite prevalence and intensity were related to temperature and humidity, which can affect the nidicolous species of ectoparasites. Residents of 2 buildings sprayed insecticides in response to Cimex sp., and this appeared to reduce ectoparasitism of S. occidentalis and C. pilosellus present at these buildings. Intensity of S. occidentalis had no influence on annual survival of big brown bats.

The effects of intraspecific and interspecific interactions on preferred questing sites of ticks, specifically nymphs and larvae of Haemaphysalis longicornis and Haemaphysalis megaspinosa, were examined in Boso Peninsula of central Japan from October 1996 to September 1999. Haemaphysalis longicornis were primarily segregated from H. megaspinosa by season. All stages of the 2 tick species preferred sedges. Three-way contingency tables and log-linear models were used to test for independence of occurrence and to quantify associations between species and stages with similar host ranges. The shifts of questing site from leaves to stem tips and from 40–49 cm to greater heights were observed in both species, which suggests that these sites are more suitable for ticks and that aggregation may serve as protection from severe conditions. In contrast, a shift to a lower height was observed in H. longicornis nymphs and larvae when other species were present, suggesting that they were driven away by other species, especially H. megaspinosa. Heterospecific clusters composed of at least 2 species were formed on stem tips more frequently. It is concluded that questing site was affected by both aggregation pattern and the presence of other species.

During a parasitological survey of Galaxias maculatus (Jenyns, 1842) in the Maullín Basin (Chilean Patagonia), specimens of a new species of Monogenea were collected from the gills. This species is described as the only member of a proposed new genus, Inserotrema n. gen. (Dactylogyridae, Ancyrocephalinae), characterized by similar hooks with 2 subunits, overlapping gonads, coiled cirrus with counterclockwise rings, articulated accessory piece formed by 2 parts, a needlelike sclerite threading the distal part of the MCO, and a sclerotized midventral vagina. This new genus is proposed for dactylogyrids from gills of galaxiids (Galaxiidae). Inserotrema puyei n. sp. infects gills of G. maculatus from Llanquihue Lake, Maullín River, and Maullín Estuary. This is the first species of Ancyrocephalinae described from gills of a galaxiid.

Spermiogenesis and the ultrastructural characters of the spermatozoon of Mesocestoides lineatus are described by means of transmission electron microscopy, including cytochemical analysis for glycogen. Materials were obtained from a golden hamster (Mesocricetus auratus) after experimental infection with tetrathyridia metacestodes obtained from naturally infected lizards (Anolis carolinensis) from Louisiana. Spermiogenesis in M. lineatus is characterized by the orthogonal growth of a free flagellum, a flagellar rotation, and a proximodistal fusion. The zone of differentiation contains 2 centrioles associated with striated rootlets and a reduced intercentriolar body. The mature spermatozoon of M. lineatus lacks a mitochondrion, and it is characterized by the presence of (1) a single, spiraled, crested body 150 nm thick; (2) a single axoneme of the 9 ‘1’ pattern of trepaxonematan Platyhelminthes; (3) a parallel and reduced row of submembranous cortical microtubules; (4) a spiraled cordon of glycogen granules; and (5) a spiraled nucleus encircling the axoneme.

The complete mitochondrial (mt) genome of the neuropathogenic bird schistosome Trichobilharzia regenti was fully sequenced in order to develop molecular markers for future diagnostic, molecular ecological, population, and phylogenetic studies. The genome was 14,838 bp in length, with a 68.4% AT bias in protein coding regions. A repeat element (3 × 184 bp) between trnV and trnW distinguished a single short noncoding region. As 9 of 14 genera of schistosomes parasitize birds, future characterization of their mt genomes is desirable for species-specific and strain- or population-specific diagnostic markers; this concerns not only the nasal representatives, e.g., T. regenti characterized in this study, but also numerous species within the predominant group of visceral (blood dwelling) bird schistosomes.

Tritrichomonas foetus is the cause of trichomoniasis in cattle. Severe infection is often associated with heavy neutrophil and macrophage accumulation, although it is not known how this response protects during early parasite colonization. The goal of this study was to examine the effects of an early host response upon initial T. foetus colonization within the murine reproductive tract. Mice depleted of neutrophils before T. foetus infection had a significantly higher parasite burden within the reproductive tract compared with mock-depleted control mice. Additionally, gp91^phox−/−/iNOS^−/− , and iNOS^−/− mice had substantially larger parasite burdens than C57BL/6 control mice, whereas gp91^phox−/− mice had similar parasite burden to C57BL/6 control mice. Interestingly, phorbol 12-myristate 13-acetate-stimulated neutrophils and macrophages isolated from all groups of mice were unable to kill T. foetus in vitro. However, macrophages isolated from gp91^phox−/− and C57BL/6 mice stimulated with interferon-γ and lipopolysaccharide were able to kill T. foetus in vitro, whereas macrophages isolated from gp91^phox−/−/iNOS^−/− and iNOS^−/− mice were unable to kill T. foetus, suggesting the ability of macrophages to produce reactive nitrogen species but not reactive oxygen species (ROS) is critical for parasite killing during early infection in vivo and in vitro. Additionally, neutrophils seem to control early dissemination of T. foetus throughout the reproductive tract, although production of ROS is not critical for this process.

Nonindigenous parasite introductions and range expansions have become a major concern because of their potential to restructure communities and impact fisheries. Molecular markers provide an important tool for reconstructing the pattern of introduction. The parasitic castrator Loxothylacus panopaei, a rhizocephalan barnacle, infects estuarine mud crabs in the Gulf of Mexico and southeastern Florida. A similar parasite introduced into Chesapeake Bay before 1964, presumably via infected crabs associated with oysters from the Gulf of Mexico, was identified as L. panopaei. Our samples of this species during 2004 and 2005 show that the introduced range has expanded as far south as Edgewater, Florida, just north of the northern endemic range limit. The nonindigenous range expanded southward at a rate of up to 165 km/yr with relatively high prevalence, ranging from 30 to 93%. Mitochondrial DNA sequences from the cytochrome oxidase I gene showed that these nonindigenous L. panopaei are genetically distinct from the endemic parasites in southeastern Florida and the eastern Gulf of Mexico. The genetic difference was also associated with distinct host spectra. These results are incompatible with an eastern Gulf source population, but suggest that unrecognized genetic and phenotypic population structure may occur among Gulf of Mexico populations of Loxothylacus.

Gregarina cubensis is an apicomplexan parasite that infects the intestinal lumen of the death's head cockroach (Blaberus discoidalis). This study evaluated the effects of 3 temperatures on the development and viability of G. cubensis. Three groups of B. discoidalis were inoculated with G. cubensis oocysts and maintained at 15, 27, and 40 C. The alimentary canal was removed from 3 cockroaches in each group every 24 hr until mature gametocysts were found in the rectum or feces, and prepared for histological examination. Gregarina cubensis establishment and development were more rapid at 15 C than at 27 C. Development of G. cubensis at 40 C did not progress beyond the intracellular stage.

Previous studies have successfully shown evidence for parasitic infections in human remains from various archaeological sites. However, in the case of Korea, since there have been very few paleoparasitological reports published, pre-20th century parasitic infection patterns remain obscure. Therefore, in order to partly fill this gap, we are reporting on a case of paleoparasitic infection from the feces of a 15th century child mummy from Yangju, Korea. In the course of the present study, we found the eggs of Clonorchis sinensis, Ascaris lumbricoides, and Trichuris trichiura in the feces of the mummy. Trichuris trichiura eggs were found in far greater numbers than other parasite eggs; in fact, intact bipolar plugs were clearly observed and even the larvae were still visible in some eggs. The eggs of C. sinensis and A. lumbricoides were also well preserved, though not in as great a number. Since we could find a number of well-preserved larvae-containing eggs, we are encouraged that successful extraction, amplification, and sequence determination of ancient DNA from the paleoparasite eggs might be possible in future studies. With additional paleoparasitological investigation using feces from Korean mummies, we hope that a history of parasite infection in Korea will be reconstructed.

Metacercariae survival patterns and their distribution in second intermediate odonate hosts were examined for 4 species of frog lung flukes. Surveys of aquatic larvae and recently emerged teneral dragonflies and damselflies indicated that prevalence and mean abundance of Haematoloechus spp. metacercariae were significantly lower in teneral dragonflies than larval dragonflies, while there was no significant difference in prevalence or mean abundance of Haematoloechus spp. metacercariae among larval and teneral damselflies. Experimental infections of dragonflies indicated that metacercariae of Haematoloechus coloradensis and Haematoloechus complexus were located in the head, thorax, and branchial basket of dragonflies, whereas metacercariae of Haematoloechus longiplexus and Haematoloechus parviplexus were restricted to the branchial basket of these hosts. Metacercariae of H. coloradensis, H. complexus, and H. longiplexus infected the head, thorax, and abdomen of damselflies, but these insects were resistant to infection with H. parviplexus. Subsequent metamorphosis experiments on experimentally infected dragonflies indicated that most metacercariae of H. longiplexus were lost from the branchial basket during metamorphosis, but most metacercariae of H. coloradensis, H. complexus, and H. parviplexus survived dragonfly metamorphosis. These observations suggest that the observed ecological host specificity of H. longiplexus in semiterrestrial leopard frogs may be due to few metacercariae of H. longiplexus reaching these frogs in a terrestrial environment. Because of the uncertain validity of Haematoloechus varioplexus as a distinct species from its synonym H. parviplexus, their morphological characters were reevaluated. The morphological data on H. varioplexus and H. parviplexus indicate that they differ in their acetabulum length and width, ovary shape, testes length, and egg length and width. Experimental infections of plains leopard frogs, northern leopard frogs, and bullfrogs with worms from bullfrogs indicate that the synonymy of H. parviplexus with H. varioplexus is not warranted, and that these flukes are distinct species, i.e., H. parviplexus in bullfrogs and H. varioplexus in plains leopard frogs and northern leopard frogs.

Dicyemid mesozoans (Phylum Dicyemida) are endoparasites (or endosymbionts) that typically are found in the renal sac of benthic cephalopod mollusks such as octopuses and cuttlefishes. Adult dicyemids likely adhere to the renal appendage of hosts via cilia of calotte peripheral cells. These cilia seem to be continuously worn away in the interaction between the dicyemids and the epidermal cells of host renal appendages. We cloned 4 cDNAs and genes, alpha-tubulin, beta-tubulin, tektin B, and tektin C, which are thought to play a key role in ciliogenesis, from Dicyema japonicum, and studied expression patterns of these genes by whole-mount in situ hybridization. We detected coexpression of these genes in the calotte peripheral cells, but not in the trunk peripheral cells. This suggests that regeneration and turnover of cilia continuously occur in the calotte. In vermiform and infusoriform embryos, we also detected coexpression patterns of these genes, which might correlate with ciliogenesis during the embryogenesis. We also predicted the secondary structure and the coiled-coil regions of dicyemid tektins.

Cryptosporidium spp., enteropathogens of humans and other animals, are members of the Apicomplexa. In parasites belonging to this phylum, proteases have been shown to play a key role in the invasion of host cells, organelle biogenesis, and intracellular survival. The subtilases constitute a family of serine proteases present in prokaryotes, eukaryotes, and viruses. The C. parvum subtilase gene, CpSUB1, encodes a transcript of 3,972 base pairs (bp) and 1,324 amino acids. Using homologous polymerase chain reaction primers, a similar gene, ChSUB1, which has 98% (4,007 bp/4,050 bp) identity to CpSUB1, was found in C. hominis. The alignment of the CpSUB1 and ChSUB1 nucleotide sequences identified primarily silent substitutions, consistent with the absence of diversifying selection. The catalytic domain of CpSUB1 is very similar to that of other Apicomplexa (>38% amino acid identity and >57% similarity) and to the bacterial subtilisin BPN from B. subtilis (36 and 47%). Transcriptional up-regulation during merozoite development was observed in cell culture, and a predicted 76-bp intron located near the 3′ end of the open reading frame was confirmed experimentally. Cryptosporidium parvum infection in cell culture was significantly inhibited by subtilisin inhibitor III and other serine protease inhibitors, emphasizing the importance of the parasite's subtilase for intracellular development and the enzyme's potential as a drug target.

We examine the charts of 408 malaria-naïve neurosyphilis patients given malaria therapy at the South Carolina USPHS facility, with daily records encompassing at least 93% of the duration of infection, and focus on the 152 patients infected with the St. Elizabeth strain of Plasmodium vivax, 82 with the McLendon strain of Plasmodium falciparum, 36 with the USPHS strain of Plasmodium malariae, and 15 with the Donaldson strain of Plasmodium ovale in whom gametocytes appeared before drug, or other, intervention. In P. vivax infections, fever and parasitemia were higher after gametocytes were first detected than before; in P. malariae infections, parasitemia was higher. In P. ovale infections, fever and parasitemia were similar before and after. In P. falciparum infections, fever, parasitemia, and fever frequency were lower after gametocytes were first detected than before. Parasitemia and temperature correlated in P. vivax infections, before and after gametocytes were first detected; parasitemia and temperature at first fever were not correlated in infections with any species. Gametocyte density correlated with parasitemia in P. malariae and sporozoite-induced P. falciparum and P. vivax infections. Fevers and detected gametocytemia coincided more often than expected by chance with P. vivax and P. ovale; fever temperature and gametocyte density were not correlated in infections with any species.

The mucus gel layer overlying the gastrointestinal epithelium plays an important role in host–pathogen interactions. The initial interaction between the coccidian parasite Eimeria tenella and host cells of the intestinal epithelium must occur across this mucus interface. In this study, we examined the relationship between E. tenella and avian mucin, in particular the effect of purified intestinal regional mucin on parasite adherence and invasion in vitro. Secreted mucin from the chicken duodenum and cecum was purified by density gradient centrifugation and gel chromatography. Parasite invasion studies were performed in the Madin-Darby bovine kidney cell model. Eimeria tenella adherence to chicken duodenal mucin was detected, whereas adherence to cecal or bovine mucin was not shown. Parasite invasion into epithelial cells was not influenced by bovine mucin, whereas chicken mucin purified from the duodenum and cecum significantly inhibited invasion. Inhibition of E. tenella invasion into cells by mucin from the duodenum was marginally greater than that of the cecum, but this was not significant. This study demonstrated E. tenella interaction with native chicken intestinal mucin, which in turn inhibited parasite invasion into epithelial cells in vitro.

Two species of Africana Tavassos, 1920 (Africana kinixysae n. sp. and Africana congoensis n. sp.) are described from Kinixys erosa (Schweigger, 1812) in the Democratic Republic of Congo. Light microscopy and scanning electron microscopy studies revealed morphological differences in the structure of the male caudal end and cephalic end, which helped to differentiate these new species from the other species of the genus, and from each other. A key for determination of all species in the genus is provided.

A new nematode species, Cucullanus angeli n. sp., is described from specimens recovered from the intestine of Vieja intermedia (Günther, 1862) from the Lacantun River, State of Chiapas, Mexico. It is characterized largely by having an unusual distinct unpaired median papilla present on anterior cloacal lip in the male and the situation of phasmids (close to eighth pair of papillae); it is further characterized by equal spicules (length 175–475 μm), distribution of caudal papillae, a preanal sucker surrounded by first and second pair of papillae, and conical tail in both sexes, ending in small terminal digitiform process (being more conspicuous in the female).

A new genus and species of Seuratiinae is described based on adults recovered from the Juan Fernández and Kermedec Petrels (Pterodroma externa and P. neglecta) from the Juan Fernández Islands of Chile (south Pacific). Navonia pterodromae n. gen., n. sp. is distinguished from the other genera and species in Seuratiinae by the combination of the following characters: (1) cordons arising from commissures of buccal lips originating a finely denticulate collarette divided into 2 lateral lobes, and detached from underlying cuticle; (2) inconspicuous simple to bicuspid deirids; (3) presence of area rugosa; and (4) presence of a large left spicule in the male. It is also distinguished from some genera and species in this subfamily by the presence of a lateral furrow. Seuratia shipleyi was also recorded from P. externa. These are the first records of helminths in these 2 hosts and the first records of Seuratiinae from Chile.

A previously unrecognized microsporidian (Kabatana newberryi n. sp.) is described from the musculature of Eucyclogobius newberryi (Gobiidae) in Big Lagoon, Humboldt County, California. Spores are ovoid, ranging in size from 2.8 ± 0.3 μm in total length and 1.9 ± 0.4 μm in width (measurements of 30 spores made by calculation from micrograph). The polar filament has 9–10 coils in 1–2 rows. Development occurs in direct contact with host muscle cell cytoplasm, without xenoma or sporophorous vesicle. Phylogenetic analysis of the new species and of 35 other microsporidians known to infect fish using 1,115 base pairs of aligned 16S rRNA gene indicate the new species is most closely related to Kabatana takedai. However, the new species differs by 11% sequence divergence from K. takedai. Divergence in morphology and genetic data allow for diagnosis from all other fish-infecting microsporidia and supports recognition of a new species of microsporidian, Kabatana newberryi n. sp., presently known only from a suspected specific host, the endangered tidewater goby Eucyclogobius newberryi.

We describe a new species of Dipetalonema occurring in the body cavity of Ateles chamek (Humboldt, 1812) from north-central Bolivia. Morphologic characters serving to separate Dipetalonema yatesi n. sp. from known forms include a vagina vera with a simple tube and thin walls and a left spicule, which possesses a handle shorter than the lamina (ratio 2.7); the latter displays an anterior membranous alae similar in length to the terminal flagellum, a distal extremity of the left spicule within a simple hook and a membrane, phasmids at the basis of the lappets, and heterogeneous muscles occupying the whole cavity. Dipetalonema yatesi n. sp. can be separated from Dipetalonema robini, Dipetalonema gracile, and Dipetalonema graciliformis, between other characters, in having a simple vagina vera instead of a sinuous one, and from Dipetalonema caudispina and Dipetalonema freitasi in having the lamina of the left spicule divided in a membranous alae and a terminal flagellum.

Cryptosporidium sp. cervine genotype could possibly emerge as an important human pathogen because current evidence suggests this genotype has a wide host range and zoonotic potential. However, there is confusion in the taxonomy of the Cryptosporidium sp. cervine genotype because different names have been used to refer to this genotype in previous studies and in sequences submitted to GenBank. Consequently, we lack a clear understanding of the epidemiology of this genotype. In the present study, the Cryptosporidium sp. cervine genotype was identified in sheep, was characterized using 3 genes (18S rDNA, COWP, and HSP-70), and was compared with data from all previous reports. Intragenotypic variations were found at the 18S rDNA gene in Cryptosporidium sp. cervine genotype with 3 different subtypes (cervine 1–3). No sequence variation at HSP-70 and COWP genes were observed. Sheep should be considered as an important reservoir for this zoonotic genotype of Cryptosporidium sp.

Haemoproteus mesnili (Bouet 1909) Wenyon 1926 is redescribed from the spitting cobra, Naja nigricollis nigricollis, of Tanzania. Mature gametocytes in the acute phase of infection averaged 17.7 × 7.3 μm, with LW 128.1 μm², and L:W ratio 2.52. Nuclei were visible in both sexes. Both sexes were heavily pigmented, with 31–62 black granules dispersed in macrogametocytes; 20–46 granules were often clumped or concentrated near ends of microgametocytes. The halteridial form was present in 28% of active-phase gametocytes, but in only 8% of those in chronic phase. A few large, possibly first generation, meronts were present in cardiac muscle; uninucleate parasites within parasitophorous vacuoles in splenic cells produced small rounded or ovoid meronts, 12.2 × 9.6 μm, with 12–16 deeply basophilic, square-to-rectangular cytomeres. Meronts with 17–32 cytomeres were 16.9 × 11.9 μm. Meronts, 20 × 16 to 26 × 22 μm, contained 51–57 cytomeres. Mature meronts were ovoid, 13.7 × 11.5 μm, with many rounded merozoites. Haemoproteus balli n. sp, found in an Egyptian cobra, Naja haje haje of Kenya, differs from H. mesnili in average gametocyte dimensions, 10.8 × 7.7 μm; LW, 83.2 μm²; L/W ratio, 1.42; absence of halteridial forms; sparse pigmentation (3–10 granules); and presence of a broad peripheral band, apparently chromatin, along one side of microgametocytes.

Haemoproteus spp., with circumnuclear gametocytes and tentatively belonging to Haemoproteus belopolskyi, are widespread and prevalent in warblers belonging to the Sylviidae, with numerous mitochondrial cytochrome b (cyt b) lineages detected among them.We sampled the hemoproteids from 6 species of warblers adjacent to the Baltic Sea. Parasites were identified to species based on morphology of their gametocytes, and a segment of the parasite's cyt b gene was sequenced. Sixteen mitochondrial cyt b lineages of hemoproteids with circumnuclear gametocytes were recorded. Two clades of lineages (clade A in species of Acrocephalus and Hippolais and clade B in species of Sylvia) with sequence divergence between their lineages >5% are distinguished in the phylogenetic tree. Within the clades A and B, the genetic distance between the lineages is ≤3.9 and ≤2.8%, respectively. We compared the morphology of gametocytes of 3 lineages (hHIICT1, hMW1, and hSYAT2) in detail. The lineages hHIICT1 and hMW1 (clade A) belong to the morphospecies H. belopolskyi. Parasites of the lineage hSYAT2 (clade B) are described as a new species Haemoproteus parabelopolskyi, which can be readily distinguished from H. belopolskyi by the significantly smaller nuclei of its macrogametocytes. Lineages closely related to H. belopolskyi and H. parabelopolskyi are identified. The sequence divergence between lineages of these 2 morphospecies ranges between 5.3 and 8.1%. It seems probable that avian Haemoproteus spp. with a genetic differentiation of ≥5% in mitochondrial cyt b gene might be morphologically differentiated at the stage of gametocytes. This study establishes the value of both PCR and morphology in identification of avian hemoproteids.

The first line drugs for the treatment of leishmaniasis are antimonial derivatives. Poor clinical response may be credited to factors linked to the host, the drug, or the parasite. We determined the sensitivity of Leishmania sp. promastigotes and amastigotes by counting parasites exposed to increasing concentrations of meglumine antimoniate (Glucantime). Leishmania braziliensis promastigotes were significantly more sensitive than those belonging to other species. The sensitivity of L. braziliensis isolates from patients with unfavorable clinical outcome, such as therapeutic failure or relapse, was significantly lower than those from patients who had clinical cure. Poor clinical response to therapy (therapeutic failure or relapse) was also associated with inadequate antimonial therapy. We also found a significant and positive correlation between promastigotes and intracellular amastigotes with regard to their in vitro susceptibilities to meglumine antimoniate. Our data provide evidence for an association between the sensitivity of promastigotes to antimonials in vitro and clinical response to therapy in American tegumentary leishmaniasis. The high sensitivity of the local L. braziliensis to meglumine antimoniate in vitro provides an explanation for the good clinical response of cutaneous leishmaniasis in the municipality of Rio de Janeiro, Brazil, even when low-dose regimens are employed.

Toxoplasma gondii is a well-recognized cause of disease in congenitally infected and immunocompromised individuals. Histone deacetylases (HDAC) comprise a family of enzymes that participate in the regulation of chromatin structure, gene expression, and cell signaling in eukaryotes. Toxoplasma gondii expresses a HDAC Class I enzyme homologous to human hdac3. Previous work showed that the histone deacetylase inhibitors (HDI) apicidin and valproic acid inhibit T. gondii infections in vitro. The present study compares the activity of hydroxamic-acid histone deacetylase inhibitors against the RH strain of T. gondii growing in HS68 human foreskin fibroblast cells. Nanomolar concentrations of suberoylanilide hydroxamic acid (SAHA), suberic bishydroxamic acid (SBHA), scriptaid, and trichostatin A (TSA) inhibited T. gondii tachyzoite proliferation. Scriptaid was the most potent hydroxamic acid inhibitor (IC₅₀ = 39 nM). In comparison, the carboxylate histone deacetylase inhibitors sodium valproate, sodium butyrate, and 4-phenylbutyrate were less potent (IC₅₀ range 1–5 mM). All of the inhibitors tested, except SBHA, completely protected the HS68 monolayers from T. gondii at concentrations 3–6 times greater than their respective IC₅₀. In contrast, nicotinamide, an inhibitor of NAD -dependent Class III HDAC, had minimal activity against T. gondii in our in vitro assays. We conclude that the hydroxamic acid class of histone deacetylase inhibitors exhibit potent anti–T. gondii activity in vitro.

The evolution of the humoral responses of IgG and IgM against 29–35-kDa Toxoplasma gondii fractions from experimentally infected goats were studied and compared by ELISA with the use of whole T. gondii soluble extracts and 29–35-kDa electroeluted proteins as antigens. The IgM response to electroeluted proteins was detected from wk 1 to wk 3 postinfection, showing a similar evolution to that observed when T. gondii crude extracts were used as antigens. These results suggest that this group of proteins could be used for a more specific detection of anti–T. gondii IgM. In the same way, the IgG response was equivalent in both cases, although when 29–35-kDa T. gondii fractions were used as antigens, the level of specific IgGs reached a peak 2 wk before than when T. gondii crude extract was used. The detection by ELISA of anti–T. gondii IgM in goats does not seem to be affected by the presence of specific IgG in serum samples when 29– 35-kDa protein fractions were used as antigens.

We describe 35 microsatellite markers from the human parasitic nematode Ascaris lumbricoides. We found 7 sex-linked markers and demonstrate that 26 autosomal loci can be scored reliably. These markers have high genetic variability and provide the tools to address multiple questions concerning the epidemiology, fine-scale genetic structure, host specificity, and mating systems of this parasite.

During a collection of ticks from vegetation in March 2006, a single adult male Rocky Mountain wood tick, Dermacentor andersoni (Stiles, 1908), was collected that exhibited unique morphological anomalies, including the absence of a leg on the right side of the body. Coxa IV on the right side also was missing in this specimen. Such teratological changes have not been reported previously for D. andersoni.

Toxoplasma gondii infection in marine mammals is of interest because of mortality and mode of transmission. It has been suggested that marine mammals become infected with T. gondii oocysts washed from land to the sea. We report the isolation and genetic characterization of viable T. gondii from a striped dolphin (Stenella coeruleoalba), the first time from this host. An adult female dolphin was found stranded on the Pacific Coast of Costa Rica, and the animal died the next day. The dolphin had a high (1:6,400) antibody titer to T. gondii in the modified agglutination test. Severe nonsuppurative meningoencephalomyelitis was found in its brain and spinal cord, but T. gondii was not found in histological sections of the dolphin. Portions of its brain and the heart were bioassayed in mice for the isolation of T. gondii. Viable T. gondii was isolated from the brain, but not from the heart, of the dolphin. A cat fed mice infected with the dolphin isolate (designated TgSdCo1) shed oocysts. Genomic DNA from tachyzoites of this isolate was used for genotyping at 10 genetic loci, including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico, and this TgSdCo1 isolate was found to be Type II.

The conventional methods for identification and typing of Leishmania species depend on previous culture isolation of the parasites. Not infrequently, culture is unsuccessful and may result in misrepresentation of the heterogeneity of the original isolate. Thus, more reliable and precise identification of genotypes of Leishmania spp. is important for a better clinical and epidemiological understanding of the disease. We evaluated the potential of LSSP-PCR targeting kDNA minicircles in discriminating different variants of the parasite with the use of clinical samples directly or cultivated parasites. The 1st step of this procedure consists of the amplification of the minicircles by conventional PCR; the 2nd step is low-stringency amplification of the minicircles previously amplified, with the use of 1 of the primers. Although LSSP-PCR produced complex and distinct kDNA signatures for isolates representing different species, further experiments demonstrated that the approach had the potential for discriminating intraspecific variants of L. braziliensis. Thus, the generated profiles were too variable to be useful as markers for species identification. Moreover, we demonstrated that the approach can be directly applied to clinical samples. In conclusion, LSSP-PCR targeting kDNA minicircles produces profiles that reflect polymorphisms of the predominant classes of minicircles, and can be useful for studies aimed at discriminating Leishmania braziliensis genotypes without the need for previous cultivation of the parasite.

The surface architecture of oocysts produced by Gregarina niphandrodes (Eugregarinorida) from Tenebrio molitor adults (Coleoptera: Tenebrionidae) as revealed by scanning electron microscopy is reported. Gametocysts were allowed to dehisce on 15-mm, round cover glasses; the cover glasses with their oocysts chains were then mounted on stubs without further processing, and sputter-coated with 20-nm gold–palladium. Scanning electron microscopy was performed at 10– 15 kV with a Hitachi 3000N SEM. Oocysts retained their characteristic shapes as reported in the original species description but showed longitudinal ridges of relatively uniform height, width, and spacing, in separate fields on either side of a central equatorial bulge in the oocysts. There was no ultrastructural evidence of an enclosing external sheath holding the oocysts in a chain. Oocyst ends were flared slightly, and the chain itself was twisted, with adjacent oocysts offset slightly from one another. This article now provides an additional set of structural characters potentially useful in gregarine systematics.

In total, 21 Hudsonian godwits, Limosa haemastica (Charadriiformes: Scolopacidae), were examined for helminths, 10 from Bristol Bay, Alaska, and 11 from Churchill, Manitoba. Seventeen species of helminths (9 trematodes, 6 cestodes, 2 nematodes) were collected, but only 1 trematode species, Plagiorchis elegans, was found in common between the 2 sample sites. All 17 species are new records for this host and 2 cestodes, Capsulata edenensis and Malika limosa, are new records for North America. In general, both prevalence and intensities were low, and species richness ranged from 1 to 6 (mean = 2.4). Most of the differences in the helminth faunas between the 2 sites were attributed to difference in habitats, freshwater in Manitoba versus saltwater in Alaska.

Ixodes (Pholeoixodes) gregsoni Lindquist, Wu, and Redner, a species of hard tick described in 1999 in Canada, was recovered from a harvested fisher (Martes pennanti Erxleben) and a domestic cat (Felis silvestris catus Ragni and Randi) in Vermont in 2001 and from harvested mink (Mustela vison Schreber) in Maine in 2003. These samples are the first records of this species within the United States. Although knowledge of this tick's natural history and distribution are still preliminary, these records indicate a possible greater distribution for I. gregsoni than initially perceived. Although its status as a disease vector is presently unknown, natural resource professionals should be aware of the possibility of this tick's occurrence in the northeastern United States.

Ixodes philipi ticks were collected from the nest burrows of streaked shearwaters, Calonectris luecomelas, on 3 different islands of Japan (Awashima: 38°45′N, 139°24′E; Mikurajima: 33°52′N, 139°36′E; and Omorijima: 36°8′N, 133°10′E). The mitochondrial cytochrome oxidase subunit I (COI) gene sequence was determined for each tick. The COI sequences of 9 other ixodid tick species also were determined, and they were used for taxonomic positioning of I. philipi. A metastriata tick, Amblyomma triguttatum, was used as an outgroup reference for the analysis. Phylogenetic examination indicated that the I. philipi ticks are on the branch with Ixodes turdus and Ixodes acutitarsus weakly, and the bootstrap value of this branching was low. Three different analyses, maximum parsimony, genetic distance, and maximum likelihood, support this conclusion. To further refine this analysis, 2,761 base pairs (bp) of sequence, which included the genes for tRNA^Met, NADH dehydrogenase subunit 2 (ND2), tRNA^Trp, tRNA^Cys, tRNA^Tyr, and COI, were determined and compared for 6 I. philipi ticks from the 3 different collection sites. Although a base substitution (T to C in the ND2 gene for an Awashima tick) and 2 transitions (G to A in the COI gene for 1 Omorijima tick) have occurred, the overall sequences were highly conserved. Preserved mitochondrial sequences in the ticks from 3 widely separated locations suggest the possibility of gene flow, which was probably accomplished by migratory seabirds.

Parasitological examination of feces was carried out for 55 patients with diagnosed colorectal cancer before chemotherapy. Except for Cryptosporidium sp., no other intestinal parasites were found in the specimens; moreover, only the patients with watery diarrhea were Cryptosporidium sp.–positive by enzyme immunoassay. Prevalence of infection in the group of patients with diarrhea (23 persons) was 43.5%, whereas it was 18% for the entire group of patients under study. Coproantigens of this parasite were detected primarily in the patients with tumors located on the left side (in the sigmoid and descending colon).

Schistosoma mansoni has a genome of 270 Mb contained on 8 pairs of chromosomes. C-banding has been a useful technique in identifying the 7 autosomal and sex chromosomes. However, even with C-banding, S. mansoni chromosomes 5, 6, and 7 are difficult to discriminate from each other, because of their small sizes, morphological similarity, and poor banding patterns. We have identified probes that specifically paint chromosomes 5, 6, and 7 of S. mansoni with the use of chromosome microdissection and the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). Exact chromosome identification is required for accurate chromosome mapping of genomic clones and genetic elements, which is an essential component of the schistosome genome project.