The work presented here deals with the status of glutathione-S-transferase (GST; E.C. 2.5.1.18), the major enzyme of the phase II detoxification pathway, in bovine filarial worms Setaria cervi. GST activity was determined in various subcellular fractions of bovine filarial worms S. cervi (Bubalus bubalis Linn.) and was found to be mainly associated with cytosolic and microsomal fractions. The respective specific activities of the enzyme from cytosolic and microsomal fractions of S. cervi females were determined to be 0.122 ± 0.024 and 0.010 ± 0.0052 μmol/min/mg protein, respectively. Cytosolic enzyme was found to possess optimal activity between pH 6.5 and 7.5, whereas the microsomal enzyme showed a broad pH optima, centered at pH 6.0. Kinetic studies on the cytosolic and microsomal forms of the enzyme revealed significant differences between them, thereby indicating that microsomal GST from S. cervi is quite distinct to the cytosolic protein catalyzing the same reaction.

The presence of trehalase and trehalose phosphorylase in L3 and L4 larvae of Anisakis simplex was demonstrated. The activity of trehalase and trehalose phosphorylase in L3 larvae was 6 and 10 times higher, respectively, than in L4 larvae. This suggests that trehalose metabolism is more important for L3 than L4 larvae. Trehalases of L3 and L4 differ in their characteristics. The enzyme of L3 was present mainly in the lysosomes and cytosol, whereas in L4 the highest enzyme activity was measured in the lysosomal fraction. Trehalase activity was increased by 29% in L3 and 55% in L4 with the addition of Mg² (0.1 mmol). Tris inhibited trehalase in L3 larvae by 42% and in L4 by 25%. The enzymes differed in their reaction to EDTA, CaCl₂, ZnCl₂, and CH₂ICOOH (all 0.1 mmol). High activity of trehalase from L₃ larvae was measured within the pH range of 5.0 to 6.5, with an optimum pH of 6.1. The trehalase was a thermally tolerant enzyme from 25 C to 60 C. The enzyme lost half of its activity after preincubation without substrate above 75 C. The paper also discusses the similarities and differences in characteristics of trehalase from A. simplex larvae and presents the comparison to enzymes from other nematodes.

The ultrastructural organization of the mature spermatozoon of the trypanorhynch cestode Parachristianella trygonis is described by transmission electron microscopy. The spermatozoon is a long and filiform cell, tapered at both ends, lacking both mitochondrion and crested bodies. Its cytoplasm contains 2 axonemes of the 9 ‘1’ pattern of the Trepaxonemata longitudinally displaced, parallel cortical microtubules, the nucleus and glycogen in form of both α-glycogen rosettes and β-glycogen particles. The anterior extremity of the spermatozoon is characterized by the presence of an arclike row of up to 10 parallel cortical microtubules that partially surround the first axoneme. The present study emphasizes the ultrastructural similarity between mature spermatozoa of all 4 trypanorhynchs that have been studied to date. Nevertheless, several features, i.e., the characteristics of spermatozoa extremities, the absence of crested bodies, and the possible presence of an arclike layer of cortical microtubules, need a more thorough analysis or confirmation in some of these species.

Schistosoma mansoni occurs in tropical regions where levels of ultraviolet B (UVB; 290-320 nm) light are elevated. However, the effects of UVB on parasite transmission are unknown. This study examines effects of UVB on the miracidia and sporocysts of S. mansoni, focusing specifically on intramolluscan development, infectivity, and the ability to photoreactivate (repair DNA damage using visible light). Histology revealed that miracidia irradiated with 861 J·m⁻² underwent abnormal development after penetrating Biomphalaria glabrata snails. Total number of sporocysts in snail tissues decreased as a function of time postinfection (PI), among both nonirradiated and irradiated parasites; however, this decrease was greater in the latter. Moreover, whereas the proportion alive of nonirradiated sporocysts increased PI, that of irradiated sporocysts, i.e., derived from irradiated miracidia, decreased. Irradiation of miracidia with UVB resulted in decreased prevalence of patent infection (defined by presence of daughter sporocysts) in a dose-dependent manner, and no infections occurred at a dose of 861 J·m⁻². Like many aquatic organisms, including the snail host, parasites were able to photoreactivate if exposed to visible light following UVB irradiation, even subsequent to penetrating snails. These photoreactivation results suggest cyclobutane–pyrimidine dimers in DNA as the primary mechanism of UVB damage, and implicate photoreactivation, rather than nucleotide excision, as the main repair process in S. mansoni.

Charlie's Pond (North Carolina) harbors a diverse community of trematodes that infect the planorbid snail Helisoma anceps. Research at the Pond began in 1984 and serves as a foundation on which to investigate long-term changes in trematode communities. In 2002, 2005, and 2006 average size and fecundity of H. anceps were calculated each month, and seasonal trends analyzed with randomization tests. Concomitantly, trematode infections were recorded, and the community composition compared to those from previous studies. Helisoma anceps in 2002, 2005, and 2006 were smaller and less fecund than snails in 1984. The trematode community was consistently diverse, with 11 species recovered in 2006 versus 7 in 1984. However, the prevalence of Halipegus occidualis was much lower than previously observed (60% in 1984) and never exceeded 20% during the latter years. The decline of emergent vegetation is likely contributing to these changes. Aquatic macrophytes increase the surface area for growth of periphyton, the food source of these snails. Limited food supplies result in lower snail growth rates and fecundity. Similarly, emergent vegetation creates foci of transmission for H. occidualis between the frog definitive host and the snail intermediate host. When these areas are lost from the Pond, probability of transmission is reduced, and prevalence in the snail declines.

Many trematode cercariae show distinct behavioral features, which have commonly been used in species identification in combination with morphological characteristics. However, information regarding cercariae behavior has often not been quantified in detail, or it is scattered in the literature, which is why the appropriate level of precision in behavioral identity, particularly in groups of cercariae species showing considerable morphological overlap, has not been properly established. In this study, we investigated one such group, the furcocercariae trematodes, by studying their behavior in a community consisting of 8 species (Diplostomum pseudospathaceum, Ichthyocotylurus variegatus, Cotylurus brevis, Cercaria spinulosa, Australapatemom sp., Australapatemom burti, Sanguinicola sp., and Bilharziella polonica) in central Finland. Our aim was not to develop an identification formula on the basis of behavior but to investigate and propose characteristic measurements applicable in separation of cercariae species. We used a 2-level approach, first recording qualitative behavioral traits of the cercariae, including swimming type and resting position; and, second, more detailed quantitative behavioral characteristics, such as resting time, swimming time, and swimming speed. Essentially, species showing a 2-phase behavior were distinguishable according to qualitative traits (resting position), whereas with those showing continuous swimming behavior, a combination of qualitative and quantitative traits (swimming speed) was required. These results suggest that characteristics of cercariae behavior can not only be used in species identification but also in general life history comparisons investigating details of the cercariae transmission.

In total, 23,750 specimens of the salmon louse, Lepeophtheirus salmonis, were collected from 3,907 juvenile pink and 3,941 chum salmon caught within the Broughton Archipelago during a 2-yr survey. The prevalence on pink salmon was significantly higher than on chum salmon in 2004 (62.3% and 58.6%, respectively) and in 2005 (26.4% and 23.1%, respectively). The mean abundance on chum salmon was significantly higher than on pink salmon in 2004 (7.0 ± 0.3 and 2.8 ± 0.2, respectively), whereas in 2005 the mean abundance did not differ between species (0.6 ± 0.1 and 0.5 ± 0.0, respectively). The mean intensity on chum salmon was significantly higher than on pink salmon in 2004 (12.0 ± 0.4 and 4.5 ± 0.2, respectively) and in 2005 (2.5 ± 0.2 and 1.7 ± 0.1, respectively). The prevalence, intensity, and abundance of L. salmonis were significantly higher on salmon belonging to both host species in 2004 compared with 2005. In both years, a majority of pink and chum salmon had 2 or fewer lice. In general, a decline in abundance of L. salmonis over the 3 collection periods in each year coincided with an increased percentage of motile developmental stages. The abundance was lowest on fish collected from zones in which the seawater surface salinity was also lowest. Seawater surface temperature was higher and salinity was lower in 2004 compared with 2005. The spatial and temporal trends in the abundance of L. salmonis in relation to host size, infestation rates, and seawater salinity and temperature, evident in both years, must be considered in future studies assessing the role of farmed salmon in the epizootiology of this parasite on juvenile salmon in this area.

Although widespread throughout the continental United States, Myxobolus cerebralis, the myxozoan parasite that causes whirling disease in salmonids, has not been reported from the state of Alaska. As part of a risk assessment for the introduction and establishment of M. cerebralis into Alaska, the distribution of the invertebrate host Tubifex tubifex was surveyed, and its genetic composition and susceptibility to the parasite were determined. Many oligochaetes, but no T. tubifex, were collected from southeastern Alaska; however, 4 lineages of T. tubifex (I, III, IV, and VI) were identified from south-central Alaska. Lineage IV had not been previously described in North America, and its susceptibility to M. cerebralis was unknown. When lineage IV T. tubifex and 3 mixed lineage (I, III, IV, and VI) groups were exposed to M. cerebralis, only lineage III became infected under our experimental conditions. Infection occurred in this lineage even when it made up just 3% of the population. Implications of the presence of nonsusceptible lineages of T. tubifex on Alaskan salmonids would be significant in areas where these lineages dominate T. tubifex populations.

We compared Heterorhabditis bacteriophora GPS11 expressed sequence tags (ESTs) to the ESTs of animal-parasitic, human-parasitic, plant-parasitic, and free-living nematodes. We identified 127 previously nondescribed ESTs of which 119 had homologs in ESTs and 8 had homologs in proteins of free-living nematodes. These ESTs were assigned putative functions in transcription, signal transduction, cell cycle control, metabolism, information processing, and cellular processes, thereby providing better insight into H. bacteriophora metabolism, sex determination, and signal transduction. We also identified 36 H. bacteriophora ESTs that had significant similarities to ESTs of parasitic nematodes, but not to ESTs or proteins of free-living nematodes species. Among these are the ESTs encoding a centrin, an ankyrin-repeat containing protein, and a nuclear hormone receptor. Our analysis also revealed that parasitic nematode-specific ESTs in this H. bacteriophora data set had more homologs in animal-parasitic nematodes than those parasitizing humans or plants.

The mechanism by which Trypanosoma cruzi egresses from infected cells at the end of the intracellular replication cycle is not understood. This study explored the role of T. cruzi–derived proteases and host-cell membrane permeability during the parasite's egress process. Treatment with a fluoromethyl ketone, known to inhibit the parasite's major protease, significantly reduced parasite egress. In addition, in the late stages of intracellular infection, cells infected with T. cruzi showed increased permeability as evidenced by dye exclusion tests. Furthermore, parasites could be antibody stained inside host cells without chemical permeabilization of the plasma membrane. These results suggest that in advanced stages of the intracellular cycle of T. cruzi, the host cells lose membrane integrity. Previous studies in our laboratory have found that antibodies present in sera of mice chronically infected with T. cruzi (antiegressin) bind the surface of infected cells and reduce parasite egress. In agreement with these reports, western blot analysis showed that several proteins in infected cell membrane extracts reacted with antibodies from infected mouse serum. The findings reported herein might have implications in the process of T. cruzi egress, as well as in the mechanism of action of antiegressin.

Eight female Peromyscus californicus were infected with 10² or 10⁴ Toxoplasma gondii culture-derived tachyzoites (Type II or X) isolated from southern sea otters. All but 2 mice survived infection and developed antibodies to T. gondii. The 2 fatally infected mice were inoculated with 10⁴ tachyzoites of the Type X strain. Parasite detection by immunohistochemistry (IHC) and DNA amplification with 2 polymerase chain reaction (PCR) methods was compared for brain, heart, lung, liver, spleen, biceps muscle, and tongue, at a mean of 41 days postinfection. Parasites were detected most commonly by IHC in spleen (8/8) and brain (6/8). DNA amplification by PCR was most successful from brain, heart, and spleen.

Cats are essential in the life cycle of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts. Samples of serum, feces, and tissues from cats from Mona, a remote island off the coast of Puerto Rico, were examined for T. gondii infection. Antibodies to T. gondii were assayed by the modified agglutination test and found in 16 of 19 (84.2%) of cats, with titers of 1:10 in 2, 1:80 in 1, 1:160 in 4, 1:320 in 3, and 1:1,280 or higher in 6. Tissues of 19 of the 20 cats were bioassayed in mice for T. gondii infection. Toxoplasma gondii was isolated from tissues of 12 cats: from the hearts of 9, skeletal muscle of 10, and brain of 1 cat. All infected mice from 10 of 12 isolates died of acute toxoplasmosis during primary infection. Genotyping of these 12 T. gondii isolates (designated (TgCatPr 1–12) by 10 multilocus PCR-RFLP markers, i.e., SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico, and the 6 multilocus microsatellite markers TUB2, W35, TgM-A, B18, B17, and M33, revealed 7 genotypes; 5 isolates had Type I alleles at all loci except at 1 microsatellite locus, and the remainder were atypical. The latter isolates of T. gondii were different biologically and phenotypically from the feline isolates from the rest of the Americas. One isolate (TgCatPr 12) was a mixed infection with 2 genotypes.

The protozoon Toxoplasma gondii has a worldwide distribution and affects many species of warm-blooded animals. In the Canadian prairies, mesocarnivores such as striped skunks (Mephitis mephitis) and raccoons (Procyon lotor) have experienced an increase in density and distribution, and they are in close contact with human dwellings. However, there has been no systematic study on the seroprevalence of T. gondii in these mesocarnivore populations. The objectives of the current project were to determine the serum antibody prevalence of T. gondii in Canadian prairie mesocarnivores and to study the relationship between antibody prevalence and species, sex, age, location, and year of collection. Antibodies to T. gondii were found in 5 of 24 (20.8%) skunks from Saskatchewan trapped in 1999 and 5 of 40 (12.5%) in 2000. Seroprevalences for T. gondii in raccoons and skunks trapped in Manitoba were 2 of 10 (20%) raccoons trapped in 2002, 7 of 44 (15.9%) trapped in 2003, and 16 of 37 (43.2%) trapped in 2004; and in 13 of 99 (13.1%) skunks trapped in 2003, 29 of 131 (22.1%) trapped in 2004, 53 of 165 (32.1%) trapped in 2005, and 30 of 51 (58.8%) trapped in 2006. Age, location, and year, but not the host species, were important variables in the determining the seroprevalence of T. gondii in skunks and raccoons. Results confirm that T. gondii is endemic in the skunk and raccoon populations in the Canadian prairies.

Neospora caninum is now recognized as a major cause of abortion in cattle worldwide, but there is no report of N. caninum infection in cattle in India. Serum samples from 427 dairy cattle and 32 dairy water buffaloes from 7 organized dairy farms located in Punjab, India, were tested for N. caninum antibodies using a commercial monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Antibodies to N. caninum were found in 35 of 427 cattle from 6 of the 7 farms; 9.6% of cows, 5.1% of heifers, and 5.0% of calves were seropositive, suggesting postnatal transmission of N. caninum on the farm. Antibodies to N. caninum were found in 16 of 32 buffaloes tested from 2 dairy farms. In total, 64 cattle and 16 buffalo sera already tested by ELISA were also evaluated by an indirect fluorescent antibody test (IFAT) to verify ELISA results. Of the 64 cattle samples, 29 sera were negative by both tests and of the 35 ELISA-positive sera, 12 had IFAT titers of 1:100 or higher (1 had IFAT titer of 100, 2 had IFAT titer of 200, and 9 had IFAT titers of 400 or higher). Of the 16 buffalo sera positive by ELISA, 1 had an IFAT titer of 1:400. Thus, antibodies to N. caninum were demonstrated in cattle sera by 2 serologic methods. To our knowledge this is the first report of N. caninum infection in cattle and buffaloes in India.

Brugia pahangi infection of dogs is a well characterized model of human lymphatic filariasis in which sera consistently show IgG or IgE reactivity to a 35-kDa antigen. Using dog lymph node B cells, we previously established a heterohybridoma cell line producing canine monoclonal IgE (cmAb 2.39) that activates and degranulates canine mast cells, and specifically recognizes a 35-kDa B. pahangi antigen. By affinity purification and sequencing of the native protein from B. pahangi adults, a 19-amino acid sequence was obtained; the derived nucleotide sequence showed homology to a Brugia malayi and 2 related Onchocerca volvulus expressed sequence tag (EST) clones from the Filarial Genome Project database. Consensus primers amplified a 244-bp product from adult and infective larval stage cDNA libraries of B. malayi, O. volvulus, and Wuchereria bancrofti, but not from those of nonfilarial nematodes. The B. malayi EST clone only showed nucleotide sequence homology to O. volvulus EST sequences. A 684-bp region from the open reading frame was expressed as a glutathione S-transferase fusion protein designated BmAl-1. CmAb 2.39, as well as serum IgE from dogs infected with B. pahangi and canine filarial heartworm, Dirofilaria immitis, recognized BmAl-1 on enzyme-linked immunosorbent assay and Western blots. BmAl-1 showed high binding affinity for a fatty acid; however, a search for sequence homology with known fatty acid binding proteins indicated that BmAl-1 is a unique fatty acid binding protein. This 35-kDa protein seems to be highly conserved in different stages and species of filarids, and it represents a previously unknown allergen that is possibly involved in the pathogenesis of filarial disease.

Various physical and chemical factors were studied to determine their effects on the viability of encysted metacercariae of Echinostoma caproni. Viability was equated with chemical excystation in an alkaline trypsin–bile salts (TB) medium. Control cysts showed excystation percentages of >90% in TB. Excystation proved to be a more reliable criterion of cyst viability than observations by light microscopy. Isolated cysts and cysts left in the snail (in situ cysts) were studied. Generally, in situ cysts proved more resistant to various physical and chemical treatments than did isolated cysts. Cysts stored for 7 days at 28 C in a Locke's 1:1 solution showed 97% excystation, suggesting that cysts of this species would survive postal delays during shipment. Of numerous marinades tested, the one that was most harmful to isolated and in situ cysts was vinegar. Isolated and in situ cysts were killed by boiling (100 C) for 1 or 3 min, but freezing at −10 C did not kill all isolated or in situ cysts after 24 hr. Concentrations of potassium permanganate ranging from 300 to 1,200 mg/L killed most isolated cysts within 5 min, but in situ cysts survived these concentrations for 24 hr. Concentrated solutions of NaCl and sucrose had no effect on the viability of isolated and in situ cysts, suggesting that their use in food preparations for molluscs would not be effective in killing echinostomatid cysts in tainted snail tissues.

The lined sea anemone Edwardsiella lineata has evolved a derived parasitic life history that includes a novel body plan adapted for life inside its ctenophore hosts. Reputedly its sole host is the sea walnut, Mnemiopsis leidyi, a voracious planktivore and a seasonally abundant member of many pelagic ecosystems. However, we have observed substantially higher E. lineata prevalence in a second ctenophore species, the ctenophore predator Beroë ovata. The interplay among these 3 species has important conservation consequences as M. leidyi introductions are thought to be responsible for the severe depletion of numerous commercial fisheries in the Mediterranean basin, and both E. lineata and B. ovata have been proposed as biological controls for invasive M. leidyi. Over a 3-yr period (2004–2006), we collected 8,253 ctenophores from Woods Hole, Massachusetts, including M. leidyi, B. ovata, and a third ctenophore, Pleurobrachia pileus, and we recorded E. lineata infection frequencies, parasite load, and parasite location. We also conducted laboratory experiments to determine the likely mechanisms for parasite introduction and the effect of each host on parasite development. We observed peak E. lineata infection frequencies of 0% in P. pileus, 59% in M. leidyi, and 100% in B. ovata, suggesting that B. ovata could be an important natural host for E. lineata. However, in laboratory experiments, E. lineata larvae proved far more successful at infecting M. leidyi than B. ovata, and E. lineata parasites excised from M. leidyi exhibited greater developmental competence than parasites excised from B. ovata. Although we show that E. lineata is efficiently transferred from M. leidyi to B. ovata when the latter preys upon the former, we conclude that E. lineata larvae are not well adapted for parasitizing the latter species and that the E. lineata parasite is not well adapted for feeding in B. ovata; these developmental and ecological factors underlie the host specificity of this recently evolved parasite.

Dicyemid mesozoans are endoparasites found in the renal sacs of benthic cephalopods. Adult dicyemids insert the distinct anterior region, termed a “calotte,” into renal tubules of the host. We cloned cDNA encoding chitinase-like protein from the dicyemid Dicyema japonicum (Dicyema-clp1), and also cloned the gene fragment corresponding to the cDNA. Dicyema-clp1 has the hydrophobic amino acid–rich region, but not the chitin-binding domains at the C terminus. Analyses using the SignalP prediction program suggest this hydrophobic amino acid–rich region is the anchor sequence to plasma membranes. The putative catalytic site in glyco18 domain exhibited 1 substitution from aspartic acid to asparagine. The gene fragment had short 9 introns (22–26 bp), and the coding sequence consisted of 10 exons (30–233 bp). Specific and strong expression of Dicyema-clp1 was detected in the calotte of vermiform stages by whole mount in situ hybridization. N-acetyl-d-glucosamine was detected on the outer surface of both peripheral cells of dicyemids and epidermal cells of host renal appendages. Dicyema-clp appears to be associated with N-acetyl-d-glucosamine in the interface between dicyemid peripheral cells and epidermal cells of the host renal appendage, and possibly aids in adhering the calotte to host epidermal cells.

The histopathology, ultrastructure, and immunohistochemistry of the alimentary canal of flounder Platichthys flesus (L.), naturally infected with the nematode Anisakis simplex s.l. (Rudolphi 1809) from the River Forth (Scotland), were investigated and described. Eight of the 16 flounders were infected with A. simplex s.l. larvae (L3); parasites were encapsulated by serosa on the external surface of the host's digestive tract (intensity of infection 1–8 parasites per host), although nematode larvae were found encysted under the peritoneal visceral serosa of the host spleen and liver and, occasionally, in the liver parenchyma (intensity of infection 3–10 parasites per host). In all sites, different structural elements were recognized within the capsule surrounding larvae. Among the epithelial cells of the intestine of 5 flounders with larvae encysted on external surface of the gut, the presence of several rodlet cells (RCs) was observed. Furthermore, often the occurrence of macrophage aggregates (MAs) was noticed in infected liver and spleen, mainly around the parasite larvae. Eight neuropeptide antisera were tested with immunohistochemistry methods on gut sections of 4 P. flesus infected with extraintestinal nematodes. Sections from the gut of 5 uninfected flounder were used for comparative purposes. In the tunica mucosa of parasitized P. flesus, several endocrine epithelial cells were immunoreactive to anti-CCK-39 (cholecystokinin 39) and -NPY (neuropeptide Y) sera; furthermore, in the myenteric plexus, a high number of neurons were immunoreactive to antibombesin, -galanin, and several to -NPY and -PHI (peptide histidine isoleucine) sera.

Interleukin 6 (IL-6) is a multifunctional cytokine that regulates various aspects of the immune response, such as acute phase reaction and hematopoiesis, and is an important signal that coordinates activities of liver cells, macrophages, and lymphocytes. Amoebic liver lesions have been studied, usually in hamsters, due to the problem of abscess development in mice. We report here the development of an experimental amoebic liver abscess (ALA) model in mice deficient in IL-6. Axenically grown amoebae were injected directly into the livers of C57BL/6 wild type (WT) and IL-6 KO^−/− mice; the abscesses produced were counted and the inflammatory process was examined on 5, 10, and 20 days postinfection. Our results showed that IL-6 KO^−/− mice develop ALA, in contrast to the WT strain, which usually do not have signs of abscess or infection. Histological analysis of the abscesses showed extended inflammatory response, mainly mediated by eosinophils, which strongly infiltrate the abscess in IL-6 K^−/− mice. The present results suggest that in mice, IL-6 could play a role in the resistance against ALA.

Tritrichomonas foetus is the causative agent of bovine trichomoniasis, a sexually transmitted disease in cattle that can result in large profit losses for cattle producers. Increasing reports have suggested that T. foetus is also the causative agent of large-bowel diarrhea in cats. To determine if the trichomonads recovered from the reproductive tract of cattle and the large intestine of cats can thrive in the same host, 2 groups of virgin Angus heifers were inoculated with T. foetus. The first group of heifers was inoculated with a bovine T. foetus isolate cultured from a naturally infected cow, and heifers in the second group were inoculated with T. foetus organisms cultured from the feces of a naturally infected cat. Over an 11-wk period, vaginal, cervical, and uterine mucus samples were analyzed, along with a single transcervical uterine biopsy sample, to determine organism and disease presence. The mucus and biopsy samples collected from each group indicate that the disease caused by feline and bovine isolates of T. foetus are comparable, but not identical.

Species of Hepatozoon are apicomplexan parasites infecting tetrapod vertebrates and hematophagous arthropods. Two species, Hepatozoon catesbianae and Hepatozoon clamatae, have been described inhabiting the erythrocytes of bullfrogs and green frogs. A number of characteristics typically used to distinguish between members of this genus are shared between these 2 species, prompting speculation as to whether or not these organisms are in fact distinct species. To test the species distinction, bullfrogs and green frogs were captured at various sites across Nova Scotia, blood samples were collected, and DNA was extracted from samples containing parasites. The internal transcribed spacer 1 (ITS-1) from geographically diverse samples of both species was amplified by PCR, sequenced, and analyzed. ITS-1 sequences from the 2 species revealed single-nucleotide polymorphisms at 6 sites. Phylogenetic analysis of these molecular data and cytopathological features place isolates of each species in separate monophyletic groups. Comparison of the ITS-1 sequences between isolates from Nova Scotia and Ontario revealed that ITS-1 sequences of H. catesbianae from a previous study were mischaracterized as being those of H. clamatae. Phylogenetic data based on molecular variation and cytopathological features from this study provide the strongest evidence to date supporting the distinction between these 2 species.

Of 2,122 marine fishes representing 36 species collected in the northeastern Pacific Ocean in the vicinity of Newport, Oregon from 1971 to 1973, 541 individuals (25.5%) representing 8 species (22.2%) were infected with hemoflagellates. Four morphologically distinct trypanosomes and 3 distinct trypanoplasms were found in fishes collected offshore, but no hemoflagellates were observed in fishes from Yaquina Bay estuary. Trypanosoma pacifica was found in English sole Parophrys vetulus, Pacific sanddab Citharichthys sordidus, and slender sole Lyopsetta exilis, and survived in 5 other species after intraperitoneal injection. Trypanosoma gargantua was found in big skate Raja binoculata, and the leech Orientobdella confluens was able to transmit the trypanosome in experimental conditions. Trypanosoma khani n. sp. occurred in P. vetulus, petrale sole Eopsetta jordani, and Dover sole Microstomus pacificus. Trypanosoma murmanense was found in L. exilis collected from 200 m, but not in L. exilis collected from 80 m. Trypanoplasma beckeri parasitized the cabezon Scorpaenichthys marmoratus. Trypanoplasma bobolsoni n. sp. was found in E. jordani, L. exilis, and P. vetulus, and survived in 2 other species after intraperitoneal injection. A distinct, but unnamed trypanoplasm, was found in P. vetulus.

Plagiocirrus loboides n. sp. (Digenea: Opecoelidae) is described from Fundulus nottii, F. dispar blairae, F. chrysotus, and Notemigonus crysoleucas from the Pascagoula River in Mississippi. Plagiocirrus loboides differs from P. primus Van Cleave and Mueller, 1932, by having a longer postcecal space (14–25% of body length vs. about 7%); a more anterior vitellarium (extending at least to the middle of the ventral sucker vs. to its posterior margin); and larger eggs (51–71 μm long by 23–34 μm wide vs. 40–55 μm long by 30–35 μm wide). Plagiocirrus loboides differs from P. testeus Fritts, 1959, by having a long postcecal space (vs. <5% of body length); irregular, oblique, contiguous testes (vs. strongly lobed, well separated, tandem testes); and a more extensive vitellarium. Plagiocirrus loboides differs from both congeners by having an ovary comprised of 3 or 4 distinct lobes rather than having an entire ovary. Plagiocirrus wuyienensis Wang, 1981, from Hemimyzon zebroidus in Fujian Province, China, is herein considered a species inquirenda because it has a Y-shaped excretory bladder. Molecular phylogenetic analysis of 28S rDNA gene fragments from P. loboides and 17 digenean species demonstrates that Plagiocirrus belongs in Opecoelidae.

The myxozoans Gadimyxa atlantica n. sp. and G. sphaerica n. sp., and G. arctica n. sp. (Myxozoa, Parvicapsulidae), are described from Gadus morhua L. and Arctogadus glacialis (Peters) (Gadidae), respectively. They develop coelozoic in bisporic plasmodia in the urinary systems. Two morphological forms of spores were found in all 3 species, i.e., wide and (sub)spherical forms. Both spore types are bilaterally symmetrical along the suture line. The wide spores, semicircular in frontal view and elliptical in apical view, have 2 spherical polar capsules, which open in the sutural or median plane mid on the flat side of the spore. Mean widths of the wide spores of G. atlantica, G. sphaerica, and G. arctica are 7.5, 10.0, and 10.0 μm, respectively. The older, more thick-walled, (sub)spherical spores with binucleate sporoplasm are 8.0, 5.3, and 7.3 μm in mean width, respectively. The mean diameters of the polar capsules of (sub)spherical spores are 2.4, 1.7, and 2.2 μm, respectively. The (sub)spherical forms of Gadimyxa are most similar to Ortholinea within the Ortholineidae, but they differ in the development of the spores and in the arrangement of the polar capsules. The polychaetes Spirorbis spp. (Spirorbidae) act as invertebrate hosts of G. atlantica. The previously described actinospores of the tetractinomyxon type develop to myxospores in Gadus morhua within 8 wk. This is the second known myxozoan 2-host life cycle in the marine environment. Phylogenetic analyses based on partial small subunit rDNA sequences places Gadimyxa spp. among Parvicapsula spp. in the Parvicapsulidae.

The heterophyid trematode Ascocotyle (Ascocotyle) felippei Travassos, 1928, is redescribed and new data on its life cycle are provided, based on types and metacercariae found in the heart bulb and gills of naturally infected guppies, Poecilia vivipara (new fish intermediate host), from a coastal lagoon in Rio de Janeiro, Brazil. Examination of the type and all voucher specimens of A. (A.) felippei collected by Travassos in the type host and locality in Brazil has shown that they possess only 32 (16 16) circumoral spines, rather than 36 (18 18) spines as previously reported. Based on the identical number and arrangement of circumoral spines, shape of the body, the presence of a long preoral lobe and posterior muscular prolongation of the oral sucker, short and wide ceca, a simple gonotyl lacking refractile bodies, and the site of infection of metacercariae (predominantly heart bulb), A. (A.) puertoricensis Price, 1932 and A. (A.) tenuicollis Price, 1935, are proposed as new synonyms of A. (A.) felippei.

Strongyloides callosciureus n. sp. (Nematoda: Rhabditoidea), from Asian sciurids, is described based on morphology, morphometry, and the small and large subunit (SSU/LSU) ribosomal RNA gene (rDNA) sequences. This new species was collected from Pallas's squirrels (Callosciurus erythraeus) in the central part of mainland Japan (Honshu), which were originally introduced from Taiwan some decades ago, and plantain squirrels (Callosciurus notatus) imported from Malaysia as personal pets. For comparison, Strongyloides robustus Chandler, 1942 was collected from American red squirrels (Tamiasciurus hudsonicus) and southern flying squirrels (Glaucomys volans) imported from the United States as personal pets. The parasitic females found in North American and Asian sciurids shared some key morphological features such as the ovary running spirally around the gut, and the shapes of the stoma in the apical view and the tail. However, morphometric features of parasitic females in North American and Asian sciurids differed significantly from each other; the former was larger than the latter, and the relative position of the vulva to the whole body length from the mouth was different. The SSU/LSU rDNA sequences supported the division of sciurid Strongyloides isolates by geographical distribution of the host and morphological features, leading us to propose the erection of new species.

We examined the phylogenetic distribution of cytochrome b haplotypes of the avian blood parasite genera Haemoproteus and Plasmodium across the migratory divide of the Swainson's thrush (Catharus ustulatus) in British Columbia, Canada. From 87 host individuals, we identified 8 parasite haplotypes; 4 of Plasmodium and 4 of Haemoproteus. Six haplotypes were novel; 1 Haemoproteus haplotype was identical to H. majoris found in the blue tit (Parus caeruleus) in Sweden, and another halotype was identical to a Plasmodium haplotype found in the white-crowned sparrow (Zonotrichia leucophrys) in Oregon. The 2 most abundant parasite haplotypes were widely distributed across the contact zone, whereas 2 other parasite haplotypes seem to have structured distributions. Compared with 74 Plasmodium and Haemoproteus haplotypes published in GenBank, haplotypes recovered from Swainson's thrushes do not form monophyletic groups, and they are closely related to haplotypes from a variety of other hosts and localities. In addition, we recovered 2 Swainson's thrush Plasmodium haplotypes from the nonmigratory orange-billed nightingale thrush (Catharus aurantiirostris) in Costa Rica. This study is the first to elucidate avian blood parasite transmission, distribution, and phylogenetic relationships in an avian contact zone in North America.

The identification of Paragonimus spp. lung flukes is based on the morphology of both the metacercaria and the adult. A very small Paragonimus sp. metacercaria was found in a freshwater crab caught in Kanchanaburi Province, West Thailand, an area where metacercariae of Paragonimus heterotremus had not been found. The metacercariae cysts were 180– 204 μm in diameter, which was smaller than metacercariae of P. heterotremus. The coefficient of difference in body size between this metacercaria and P. heterotremus was 1.69, which was greater than a subspecific difference. Adults recovered from the lungs of a cat fed with the metacercariae were morphologically similar to, but smaller than, P. heterotremus dissected from the lungs of a feline experimental host. The tegumental spines of the worm in this study were singly spaced in arrangement, which is similar to, but larger than, the spines of P. heterotremus. Therefore, Paragonimus pseudoheterotremus is proposed as a new species.

In the present study, inhibitory effects of several sulfated and nonsulfated glycoconjugates were evaluated on the in vitro asexual growth of Babesia bovis. Among the selected sulfated glycoconjugates, dextran sulfate, heparin, heparan sulfate, fucoidan, and chondroitin sulfate B strongly inhibited the parasitic growth, and all but chondroitin sulfate B induced a significant accumulation of extracellular merozoites in culture. In contrast, chondroitin sulfate A, keratan sulfate, and protamine sulfate, as well as nonsulfated dextran and hyaluronic acid, did not influence the growth. These findings indicate that the asexual growth of B. bovis merozoites is inhibited by specific sulfated glycoconjugates, possibly providing us with an important insight into the molecular interaction(or interactions) during the process of the erythrocyte invasion by B. bovis merozoites.

New and efficient methods to screen antibiotics are needed to counter increased antibiotic resistance in pathogens and the emergence of new diseases. Here we report a new insect model for screening antibiotics in vivo using the grasshopper Romalea microptera. The system is inexpensive, efficient, and flexible, avoids animal-welfare problems, and can be used to test against most major pathogenic groups. We employed this system to test 11 commercial antibiotics against a pathogenic Encephalitozoon species (Microsporidia). Oral treatment with fumagillin or thiabendazole significantly reduced pathogen spore counts, whereas spore counts of grasshoppers fed with albendazole, ampicillin, chloramphenicol, griseofulvin, metronidazole, sulfadimethoxine, or tetracycline were not significantly different from the infected controls. Quinine produced a distinct, but nonsignificant, reduction in spores, and streptomycin a nonsignificant increase in spores. Although 2 antibiotics significantly reduced spore counts, in no case was the pathogen totally eliminated. This study demonstrates the validity of this system as a method to screen antibiotics. It also corroborates the difficulty researchers and physicians have had in treating microsporidia infections, and suggests that quinine and related alkaloid compounds should be further examined as possible therapeutic agents against this group of ubiquitous pathogens. In addition, streptomycin and related compounds should be tested to determine if this widely used antibiotic enhances microsporidiosis.

Administration of tamoxifen (an antiestrogen) produced an 80% parasite load reduction in female mice, and a weaker effect of 50% in male mice. This protective effect was associated in both sexes, with an increase in the mRNA levels of interleukin (IL)-2 (a cytokine associated with protection against cysticerci) and IL-4 (no effect on infection). tamoxifen treatment modified 17-β estradiol production in females, whereas serum testosterone was not affected. However, the expression of the 2 types of estrogen receptor (ER), i.e., ER-α and ER-β, in the spleen of infected mice of both sexes, was decreased by tamoxifen treatment. In vitro, treatment of Taenia crassiceps with tamoxifen reduced reproduction and loss of motility. These results indicate that tamoxifen treatment is a new therapeutic possibility to treat cysticercosis, because it can act at both ends of the host–parasite relationship, i.e., by increasing the cellular immune response protective against the parasite and by directly affecting the parasite's reproduction and survival.

Hormones play a significant role in murine Taenia crassiceps cysticercosis, and they may also participate in the susceptibility to Taenia solium cysticercosis. In the present study, in vitro effects are reported for human chorionic gonadotropin (hCG) on the larval stages of T. crassiceps (WFU strain) and T. solium. hCG effectively promotes parasite reproduction, i.e., it increases the number of buds on T. crassiceps cysticerci and the percentage of evagination and parasite length in T. solium. This is the first report in which a direct effect of hCG is reported for a parasite. hCG or mouse luteinizing hormone could be recognized by the cysticerci as mitogenic factors and contribute to the female and pregnancy bias toward susceptibility to T. crassiceps and T. solium cysticercosis, respectively.

This article reports the first national serological prevalence of Toxoplasma gondii in India. In total, 23,094 serum samples were tested for T. gondii IgG and IgM antibodies with the use of a solid-phase immunocapture ELISA. Antibodies (IgG) were found in 24.3%; IgM antibodies were detected in 2% of the samples. The lowest seroprevalences were in the northern parts of India, with the highest in the south. These data probably reflect the effects of significantly drier conditions and, therefore, a negative impact on the survivability of T. gondii oocysts.

In this study, we recovered Schistocephalus solidus plerocercoids from singly and multiply infected three-spine sticklebacks, Gasterosteus aculeatus, transferred them individually to in vitro culture conditions, and quantified their lifetime egg output. We found a significant difference in the relationships between plerocercoid mass and lifetime egg output for parasites recovered from singly and multiply infected sticklebacks. Although egg output was strongly and positively related to plerocercoid mass amongst worms from singly infected fish, for those recovered from multiply infected sticklebacks the relationship was marginally nonsignificant and negative, with small worms achieving high levels of egg production. We suggest 2 hypotheses that may explain differences in the egg production of plerocercoids from singly and multiply infected fish. One possibility is that smaller plerocercoids in asymmetric multiple infections develop precocially, in response to host manipulation strategies of larger worms that decrease survival prospects of the host. Alternatively, small worms in singly infected sticklebacks may be prevented from becoming sexually mature because they face energetic constraints associated with having to overcome the host's immune response alone. We discuss our results in terms of recent studies examining strategic egg production in helminths.

During 2005–2006, sera and tissues from raccoons (Procyon lotor), coyotes (Canis latrans), and skunks (Mephitis mephitis) from the state of Wisconsin were tested for Toxoplasma gondii infection. Antibodies to T. gondii were found in 32 of 54 (59.2%) raccoons, 18 of 35 (51.4%) coyotes, and 5 of 7 (71.4%) skunks using the modified agglutination test and a cut-off titer of 1:20. Pooled tissues (brains, hearts, and tongues) from 30 raccoons, 15 coyotes, and 1 skunk were bioassayed for T. gondii infection in mice or cats. Viable T. gondii was isolated from 5 of 30 (16.7%) raccoons, 6 of 15 (40.0%) coyotes, and the skunk. Genetic characterization of the 12 parasite isolates by multilocus PCR-RFLP markers revealed 6 different genotypes including 5 atypical and 1 archetypal II lineages. The results indicate the prevalence of T. gondii in wildlife mammals is high and that these animals may serve as an important reservoir for transmission of T. gondii.

The prevalence of Toxoplasma gondii antibodies in 80 domestic cats was studied in the city of Colima, Mexico, using an indirect IgG-ELISA. Antibodies were found in 28.8% of the cats, with significantly higher (P = 0.029) prevalence in southern and central zones (33.8%) than the northern zone (6.6%). Prevalence among cats fed with homemade food was higher than those eating commercial pellets (40.6% [vs.] 20.8%; P = 0.055). Overall, the prevalence of T. gondii antibodies in the cats of Colima was lower than in many other countries.

The range-wide population of American woodcock Scolopax minor has been in slow, steady decline since the late 1960s. The parasite load carried by woodcock and its possible role in the population decline has not been investigated since the early 1970s. A survey of parasites in American woodcock in Connecticut was undertaken in 2002; Sarcocystis spp. was found in 32 of 78 (42%) individuals examined. Elongate sarcocysts, 25 × 125 μ, containing numerous packed bradyzoites with distinct, tightly packed villar projections of the cyst wall, were found scattered throughout skeletal type and myocardial muscle. Sarcocystis spp. was also recorded during the earlier surveys and considered common, but was not examined with the use of electron microscopy. The present study includes the first ultrastructural description of Sarcocystis sp. in the skeletal muscle of woodcock and will serve as a basis for future comparisons in woodcock.

In the present study, we report a case of hyperparasitism in Amblyomma rotundatum. During examination of live ticks immediately after collecting them from Boa constrictor snakes held in a reptile facility in Mossoró, RN, northeastern Brazil, 1 unengorged tick female was seen attached to the venter of a partially engorged female. The hypostome and chelicerae of the unengorged female had penetrated the integument of the partially engorged female to the level of the basis capitulli and the palps were splayed outward. To our knowledge, we present the second report of hyperparasitism for the genus Amblyomma.

From 2004 to 2006, 511 raccoons collected by the United States Department of Agriculture Wildlife Services as part of a study to evaluate the effects of mesomammalian predator removal on bobwhite-quail reproduction, were examined for the presence of Gnathostoma procyonis, a nematode that parasitizes the stomach of its definitive host, the raccoon. One hundred forty-one raccoons (27.6%) contained G. procyonis, with males being equally infected (27.5%) as females (27.8%). Mean intensity did not differ significantly between sexes (males—mean = 2.88, range 1–17; females—mean = 2.78, range 1–10) or between collection sites. A significant seasonal prevalence of G. procyonis was noted with a peak in March (43.3%, n = 120) and a minimum in September (5.4%, n = 37). Both collection sites exhibited significant seasonal decreases in G. procyonis, but were not significantly different from each other. There was no cumulative seasonal pattern in mean intensity of G. procyonis, and no seasonal pattern at either collection site. Host weight was not related to either G. procyonis prevalence or mean intensity. Host removal did not have an apparent effect on prevalence or mean intensity of G. procyonis.

Species of Cryptosporidium infect a broad variety of animals. Because morphological features of the secreted oocysts are not useful in identifying the parasite at the species level, molecular tools were used to accomplish this task, leading to discovery of new Cryptosporidium species. With the use of this approach, Cryptosporidium bovis has recently been described as a new species infecting bovines and several other hosts, but clearly distinct from C. parvum. In this report, we present a description of a Cryptosporidium sp. isolate from a newborn lamb from a farm in Spain. The isolate seemed to be very similar to C. bovis based on the analysis of the gene that codes for the 18S rRNA.

Three species of flying fox (Pteropus hypomelanus, P. vampyrus, and P. lylei) from Malaysia and Vietnam were screened for apicomplexan parasites by thin blood smears and polymerase chain reaction. Only 1 of 16 bats sampled from 3 localities in southeast Asia was found to be infected (P. hypomelanus from Pulau Pangkor, Malaysia). We observed micro- and macrogametocytes, with morphology consistent with Hepatocystis sp. parasites, using light microscopy. Phylogenetic analysis of the cytochrome b gene showed that the parasite from P. hypomelanus groups with 2 published sequences from Hepatocystis spp., including one from Cynopterus brachyotis, another fruit bat in the Pteropodidae.

From May 2003 to April 2004, blood samples from 395 feeder pigs in Taiwan were examined for antibodies to Toxoplasma gondii using the latex agglutination test; antibodies (titer 1:32 or higher) were found in 10.1% of 395 pigs. The results indicate a high prevalence of infection in pigs in Taiwan destined for human consumption.

Surveys on the prevalence of various intestinal parasitic infections in different geographic regions is a prerequisite for developing appropriate control strategies. The aim of the present study was to determine the prevalence of intestinal parasitic infections in schoolchildren enrolled in various schools in Srinagar City, Kashmir, India, and to assess epidemiological factors associated with the extent of endemic disease. Stool samples were collected from 514 students enrolled in 4 middle schools. The samples were processed with the use of both simple smear and zinc sulphate concentration methods, and then microscopically examined for intestinal parasites. Of the 514 students surveyed, 46.7% had 1, or more, parasites. Prevalence of Ascaris lumbricoides was highest (28.4%), followed by Giardia lamblia (7.2%), Trichuris trichiura (4.9%), and Taenia saginata (3.7%). Conditions most frequently associated with infection included the water source, defecation site, personal hygiene, and the extent of maternal education. The study shows a relatively high prevalence of intestinal parasites and suggests an imperative for the implementation of control measures.