Individual Arctic charr (Salvelinus alpinus) from Fjellfrøsvatn, northern Norway, could be categorized by their stomach contents as zooplanktivores or benthivores. Feeding specialization among these fish was evident from negative correlations between helminths transmitted by pelagic copepods (Diphyllobothrium dendriticum and D. ditremum) and those transmitted by the benthic amphipod Gammarus lacustris (Cystidicola farionis and Cyathocephalus truncatus). Occurrences of parasite species acquired from the same types of invertebrate were positively correlated in the fish. Strong relationships among habitat use, diet, and helminth infections among the Arctic charr indicated persistent foraging patterns involving long-term habitat use and feeding specialization. The distribution of all parasite species was highly aggregated in the fish samples, measured by the exponent k of the fitted negative binomial distributions (range: 0.5–7.5) and the variance-to-mean ratios (s²/x̄, range: 5–85). Charr specializing on either copepods or Gammarus predominantly contributed to high-intensity class intervals within the overall frequency distributions of the corresponding parasite species. Such fish had low infection intensities of helminths transmitted by other prey organisms. The detailed analyses of the parasite frequency distributions for fish with different habitat or feeding preferences evidently show how heterogeneity in trophic behavior contributes strongly to the commonly observed aggregation of helminths among hosts under natural conditions.

This study examined the possible involvement of cyclic adenosine monophosphate (cAMP) in the control of ciliary action of Schistosoma mansoni miracidia. Miracidia immobilized in hypertonic NaCl solution were treated with 3 compounds that are known to increase intracellular cAMP concentrations. Forskolin, at a concentration of 50 μM, induced 50.1% of the miracidia to swim in hypertonic solution. The corresponding values obtained for 3-isobutyl-1-methylxanthine (IBMX) at 1 mM and 8-bromo-cAMP at 10 mM were 42.2 and 50.4%, respectively. The motility-enhancing effect of these compounds was dose dependent. Nevertheless, the swimming speed of miracidia activated in this way was only 10% of that observed in artificial pond water (APW). Cholera toxin had no apparent effect on miracidia swimming in hypertonic NaCl solution. Likewise, swimming in APW treated with forskolin at 50 μM, IBMX at 1 mM, or 8-bromo-cAMP at 10 mM did not induce any apparent change in motility. Miracidia swimming in APW were then treated with 3 compounds that decrease the intracellular concentration of cAMP. MDL-12,330A, at a concentration of 250 μM, caused a dramatic decrease in swimming over a period of 1 hr. Likewise, SQ22536 and imidazole, at concentrations of 20 and 50 mM, respectively, caused 36.5 and 73.4% decreases in swimming under the same conditions. Finally, inhibitors of cAMP-dependent protein kinase, i.e., PKI(14-22)amide, H89, and H88, completely inhibited miracidia swimming in APW at concentrations of 25, 50, and 100 μM, respectively. These results suggest that cAMP and cAMP- dependent protein kinase are involved in osmosis-controlled ciliary motion of schistosome miracidia.

The tissue distribution, course of secretion, and sex differences of morphine were delineated in Ascaris suum. Nitric oxide (NO) release in various tissues in response to morphine and its metabolite morphine-6-glucuronide (M6G) were also examined. Ascaris suum of both sexes along with their incubation fluid were analyzed for morphine concentrations by high- performance liquid chromatography (HPLC) over a 5-day period. Various tissues were also dissected for HPLC and NO analysis. Morphine was found to be most prevalent in the muscle tissue, and there is significantly more morphine in females than males, probably because of the large amounts present in the female uterus. Morphine (10⁻⁹ M) and M6G (10⁻⁹ M) stimulated the release of NO from muscles. Naloxone (10⁻⁷ M) and N-nitro-l-arginine methyl ester (10⁻⁶ M) blocked (P < 0.005) morphine-stimulated NO release from A. suum muscle tissue. d-Phe-Cys-Tyr-d-Trp-Om-Thr-Pen-Thr-NH₂ (CTOP) (10⁻⁷ M) did not block morphine's NO release. However, naloxone could not block M6G-stimulated NO release by muscles, whereas CTOP (10⁻⁷ M) blocked its release. These findings were in seeming contradiction to our earlier inability to isolate a μ opiate receptor messenger RNA by reverse transcriptase–polymerase chain reaction using a human μ primer. This suggests that a novel μ opiate receptor was possibly present and selective toward M6G.

The reinvestigation of spermiogenesis in the proteocephalidean cestode Proteocephalus longicollis (Zeder, 1800) provides new details that were not described previously for this species. The 2 centrioles with roots are oriented at the first tangential to the long axis of the nucleus in a differentiation zone. They come to be arranged in the same plane only when a single intercentriolar body (ICB) appears between them. The ICB had been described previously as consisting of a number of dense elements (Swiderski, 1985) but, in the present study, appeared as a single, electron-dense plate. Also new for this species is the observation of 2 arching membranes that appear at the base of the differentiation zone only when the 2 flagella rotate toward the median cytoplasmic protrusion. Moreover, the formation of a crested body is described for the first time in P. longicollis. The present data support the suggestion that the process of spermiogenesis in proteocephalidean tapeworms corresponds in most features to that observed in some members of the unrelated tetraphyllidean clades, the Onchobothriidae and Phyllobothriidae. This may represent more evidence regarding the close relationship between tetraphyllidean and proteocephalidean cestodes, but existing information is fragmentary and controversial in some characters.

The present paper describes the spermiogenesis and the ultrastructure of the spermatozoon of Fasciola gigantica, as revealed by transmission electron microscopy. Spermiogenesis in F. gigantica begins with the formation of a differentiation zone containing 2 centrioles with associated striated roots and an intercentriolar body between them. Each centriole develops a flagellum. Proximodistal fusion of these flagella with the median cytoplasmic extension gives rise to the spermatozoon. Spermiogenesis in F. gigantica is characterized by the formation of a dorsolateral cytoplasmic expansion, an external ornamentation of the cell membrane, and spinelike bodies. These 3 structures were also observed in the anterior part of the spermatozoon. Our study describes for the first time the simultaneous presence of dorsolateral cytoplasmic expansion, external ornamentation of the plasma membrane, and spinelike bodies in the spermatozoon of a trematode.

The community structure and seasonal dynamics of 16 helminth species infecting green (Lepomis cyanellus) and bluegill (L. macrochirus) sunfishes in Charlie's Pond, North Carolina, was examined. One hundred and fifty-four fishes including 90 green sunfish and 64 bluegill sunfish were collected between March and November 2000 and examined for the presence of helminth parasites. Five of these species underwent significant changes in abundance in green sunfish infracommunities, 3 of which also displayed seasonal changes in prevalence. Three of the 16 species fluctuated seasonally in bluegill infrapopulations; 2 also underwent changes in prevalence. Species richness and diversity varied across the 9-mo period for both host species, whereas total helminth abundance remained constant. Analysis of component communities revealed differences in community structure for the 2 host species. Bluegills were found to harbor larger and more diverse communities. Bluegills also contained larger infrapopulations of 5 species, whereas green sunfish had greater abundance of 2 species. Interpretation of these data suggests that host species and size are strongly associated with the predictability of community structure.

This study analyzed the influence of several abiotic and biotic variables on the distribution of digenetic trematode infections in a mudsnail, Hydrobia ventrosa, population inhabiting 12 ponds on the Melabakkar salt marsh in Iceland, the northwestern limits of the geographical distribution. Nine trematode species were found to infect the snail population, which included Microphallus pirum, Microphallus breviatus, Microphallus claviformis, Maritrema subdolum (Microphallidae), Cercaria Notocotylidae sp. 11 Deblock, 1980, C. Notocotylidae sp. 12 Deblock, 1980, C. Notocotylidae sp. 13 Deblock, 1980 (Notocotylidae), Cryptocotyle concavum (Heterophyidae), and Psilostomum brevicolle (Psilostomatidae). Correlations between biotic variables (snail density in the ponds and vegetation cover), abiotic variables (distance of each pond from the sea, pond elevation above chart datum, size, average depth, salinity, and some characters of the littoral zone and sediments), and trematode infections were analyzed. These variables indirectly affected the trematode infections because some determined how attractive the ponds were for the final hosts, which were various species of marine and shore birds. We propose that their habitat use and defecating habits are the main determinants of the trematode distribution in the area.

Populations of a host species may exhibit different assemblages of parasites and other symbionts. The loss of certain species of symbionts (lineage sorting, or “missing-the-boat”) is a mechanism by which geographical variation in symbiont assemblages can arise. We studied feather mites and lice from Australian brush-turkeys (Aves: Megapodiidae: Alectura lathami) and expected to observe geographical structuring in arthropod assemblages for several reasons. First, because the brush-turkey is a sedentary ground-dwelling bird, we predicted that geographically close host populations should share more similar arthropod assemblages than distant ones. Second, because brush-turkeys do not brood their young, vertical transfer of arthropods is unlikely, and brush-turkeys probably acquire their mites and lice at social maturity through contact with other birds. Young birds could disperse and found new populations without carrying complete sets of symbionts. We predicted that young birds would have fewer species of arthropods than older birds; in addition, we expected that males (which are polygynous) would have more species than females. Birds were sampled from 12 sites (=populations) along the east coast of Queensland, Australia, that were separated by a distance of 12.5–2,005 km. In total, 5 species of mites from the Pterolichidae and 1 species from the Ascouracaridae were found. Two species of lice were collected but in numbers too low to be statistically useful. Differentiation of mite assemblages was evident; in particular, Leipobius sp. showed 100% prevalence in 3 host populations and 0% in the remaining 9. A dendrogram of brush-turkey populations based on mite assemblages showed 2 geographically correlated clusters of sites, plus 1 cluster that contained 2 sites near Brisbane and 1 approximately at a distance of 1,000 km. There was no strong effect of host age or sex on number of mite species carried. Horizontal transfer of feather mites by hippoboscid flies, in addition to physical contact between hosts, may play a role in homogenizing symbiont assemblages within populations.

Toxoplasma gondii isolates can be grouped into 3 genetic lineages. Type I isolates are considered more virulent in outbred mice and have been isolated predominantly from clinical cases of human toxoplasmosis, whereas types II and III isolates are considered less virulent for mice and are found in humans and food animals. Little is known of genotypes of T. gondii isolates from wild animals. In the present report, genotypes of isolates of T. gondii from wildlife in the United States are described. Sera from wildlife were tested for antibodies to T. gondii with the modified agglutination test, and tissues from animals with titers of 1:25 (seropositive) were bioassayed in mice. Toxoplasma gondii was isolated from the hearts of 21 of 34 seropositive white- tailed deer (Odocoileus virginianus) from Mississippi and from 7 of 29 raccoons (Procyon lotor); 5 of 6 bobcats (Lynx rufus); and the gray fox (Urocyon cinereoargenteus), red fox (Vulpes vulpes), and coyote (Canis latrans) from Georgia. Toxoplasma gondii was also isolated from 7 of 10 seropositive black bears (Ursus americanus) from Pennsylvania by bioassay in cats. All 3 genotypes of T. gondii based on the SAG2 locus were circulating among wildlife.

Calreticulin (CRT) is a calcium-binding protein and has many functions in eukaryotic cells. CRT is possibly involved in parasite host immune system evasion. To better understand the molecular basis of CRT in ticks, we cloned and sequenced 4 full-length complementary DNAs (cDNAs) from the hard tick species, Dermacentor variabilis, Haemaphysalis longicornis, Ixodes scapularis, and Rhipicephalus sanguineus, using the technique of rapid amplification of cDNA ends. The deduced amino acid sequences share high identities (between 77 and 98%) with 3 known tick CRT sequences. The major characteristics of known CRTs are observed in all 4 of our deduced tick CRTs. These include 3 major domains, a signal peptide sequence at the beginning of the coding region, 2 triplets of conserved regions, cysteine sites providing disulfide bridges for N-terminal folding, and a nuclear localization signal. Remarkably, the replacement of the endoplasmic reticulum retention signal KDEL by HEEL, which is believed to be associated with secretion of CRT into the host during feeding and was previously recorded only in 2 ticks and a hookworm, is also present in all 4 of our tick putative CRTs. In addition, the CRT gene is potentially useful for tick phylogenetic reconstruction.

Taenia crassiceps cysticerci form large infrapopulations that persist in the tissues of their rodent hosts. Early infrapopulation growth appears inhibited and is followed by rapid increases that appear not to be controlled by the host immune response. This investigation was undertaken to examine the infrapopulation growth dynamics of a normally developing strain (WFU) of T. crassiceps during a 60-day primary intraperitoneal (i.p.) infection. Three, 6, 9, 14, 28, and 60 days after i.p. inoculation of 5 cysticerci, mice were killed, and the numbers of larvae, developmental stage, and buds per larva were recorded. Larval infrapopulation abundance increased exponentially beginning on day 6 postinoculation (PI), indicating an initial lag in reproduction. A stage-structured exponential growth model, assuming no mortality, fits the larval infrapopulation dynamics in terms of the numbers of larvae in reproductive and nonreproductive stages, indicating that cysticerci evade or suppress (or both) host immune mechanisms that are parasite restrictive after the first week of infection.

Trypanosoma cruzi is the causative agent of Chagas disease, which is characterized by acute and chronic phases. During the former, parasitemia rises dramatically, then decreases significantly during the chronic phase. Immune mechanisms responsible for the parasitemia reduction have not been thoroughly elucidated. The goal of the present study was to further characterize the immune response during chronic infection. Previously, we described antiegressin, an antibody in sera from chronically infected mice. The in vitro presence of antiegressin inhibits parasite egress from infected host cells. Antiegressin appears by day 14 of an in vivo infection and is maintained through at least day 280 postinfection. The in vitro functional activity of antiegressin is initiated late in the 4–6 days intracellular growth cycle of T. cruzi; antiegressin may be added at day 4, inhibiting parasite release at day 5. Immunocytochemical staining using antineuraminidase demonstrates the presence of mature parasites inside host BALB/c fibroblasts grown in the presence of antiegressin. These results demonstrate the ability of antiegressin to inhibit emergence of developmentally mature trypomastigotes from infected host cells late in their intracellular growth cycle. We believe this antibody plays an important and novel role in achieving the low-parasitemia characteristic of chronic Chagas disease.

Suspensions of miracidia and cercariae of Schistosoma mansoni were subjected to repeated freeze–thaw cycles and then injected into resistant Salvador strain Biomphalaria glabrata snails. A pronounced increase in the number of mitotic figures, relative to uninjected, sham-injected, or diluent (water)–injected controls, was observed in the amebocyte-producing organ (APO) at 3 days postinjection (PI). After centrifugation of miracidia freeze–thaw extract (FTE), the resulting supernatant (FTS) and pellet possessed equal stimulatory activity that was approximately half that seen with FTE. Ultracentrifugation of miracidia FTS resulted in a supernatant that retained full activity, indicating a soluble molecule. Heat treatment of miracidia FTE reduced but did not eliminate activity, suggesting a nonprotein active component. Concentration or dilution of FTS by a factor of 10 gave a nonlinear dose–response relationship. Susceptible NIH albino snails injected with miracidia FTE had increased mitotic activity in the APO, which was much less than that seen in Salvador snails, whereas injection of miracidia FTE into Helisoma duryi had no discernable effect. Measurement of mitotic activity as a function of time PI showed no increase in numbers of mitotic figures in the APO at 18 hr but a large increase at 24 hr PI. Mitotic activity returned to preinjection levels by 96 hr PI, although a subsequent increase occurred at 120 hr PI.

Cercaria yamagutii Ito, 1957, was found in the marine mesogastropods Lunatia fortuni and Glossaulax didyma from the tidelands of Simpo located at the estuary of the Mankyoung River, which runs to the western coast of Korea. Metacercariae were found in a marine bivalve Mactra veneriformis after being infected with C. yamagutii experimentally. When a sea gull, Larus crassiostris, was fed with the metacercariae collected from the infected M. veneriformis, adult worms were recovered 10 days later. It was confirmed that the parasites collected from L. crassiostris were Acanthoparyphium tyosenense Yamaguti, 1939. From the results of this life cycle study, it was determined that the first intermediate hosts of A. tyosenense are L. fortuni and G. didyma. The second intermediate and final hosts are M. veneriformis and L. crassiostris, respectively. Mactra veneriformis was experimentally infected with C. yamagutii isolated from L. fortuni and G. didyma by maintaining them in a water tank for 30 min at about 20 C. The cercariae entered M. veneriformis through their incurrent siphons. Five hours after infection, the cercariae tails began to separate from the bodies, and the cercariae formed cysts. Mature cysts were formed 340 hr (14 days) after infection and identified as the metacercariae of A. tyosenense. The prevalence of A. tyosenense metacercariae was 99.5% in naturally infected M. veneriformis. This is the first report of C. yamagutii as the cercaria of A. tyosenense, and the complete life cycle of A. tyosenense was established in Korea.

The relationship of fish age class to parasitism by Paulisentis missouriensis was determined by sampling at least 29 creek chubs, Semotilus atromaculatus, from Easly Creek, Richardson County, Nebraska, every month from February 1996 to March 1997. In general, mean abundance and prevalence of the acanthocephalans increased with the age or length of chubs. It is unlikely that this distribution is explained by increased consumption of intermediate hosts by older, larger fish or by predatory fish acquiring parasites from paratenic hosts. The intermediate host for P. missouriensis is the cyclopoid copepod Acanthocyclops robustus, and creek chubs do not consume more microscopic crustaceans as they age or grow. Instead, the percentage of fish in the diet of creek chubs increases. Furthermore, P. missouriensis apparently does not use paratenic hosts. In laboratory infections, P. missouriensis survived predation of its original definitive host and transferred to the predator. Postcyclically transmitted P. missouriensis survived at least 14 days in the intestine of creek chubs, where they localized around the first flexure beyond the stomach. All stages of development of both sexes were transferred successfully. Postcyclic transmission is a plausible explanation, in some cases, for the greater worm burden frequently observed in older, larger hosts and for the occurrence in top carnivores of parasites not known to have paratenic hosts. This method of transmission appears to result in distribution of acanthocephalans to groups of animals that otherwise would be inaccessible.

Life-cycle studies were conducted on 2 molecular strains of Mesocestoides tapeworms that represent different evolutionary lineages (clades A and B). Wild carnivores, reptiles, and rodents were examined for tapeworm infections at 2 enzootic sites: (1) San Miguel Island (SMI), a small island off the coast of southern California and (2) Hopland Research and Extension Center (HREC), a field station in northern California. Results indicate that deer mice (Peromyscus maniculatus) and coyotes (Canis latrans) may play an important role in the life cycles of Mesocestoides (clades A and B) in California. Over half the coyotes at HREC and at least a third of the population of island fox (Urocyon littoralis) at SMI were found to harbor clade A adult Mesocestoides spp. One of every 4 Mesocestoides-infected coyotes had tapeworms representing both clades A and B. Experimental inoculations revealed that proglottids (clades A and B) were not directly infectious to rodents, reptiles, or dogs. On the other hand, mice, lizards, and hamsters fed tetrathyridia of Mesocestoides spp. (clades A or B) developed peritoneal tetrathyridial infections. A dog that was fed tetrathyridia (clade B) developed an adult tapeworm infection. Acephalic metacestodes given orally to western fence lizards, laboratory mice, or domestic dogs did not result in metacestode or adult tapeworm infections. Whereas most clade A acephalic metacestodes from dogs were asexually proliferative, clade A tetrathyridia isolated from wild deer mice did not show evidence of asexual replication. Our study supports the hypothesis that a second, as of yet unidentified, intermediate host is necessary to complete the life cycles of Mesocestoides spp., and that acephalic metacestodes represent an aberrant form, incapable of further development.

Sexual transmission is a widespread means of infection, but apart from those in humans, the ecology of sexually transmitted organisms is not well known. In this study, we present an ecological study of a sexually transmitted mite, Parobia husbandi Seeman and Nahrung (Acari: Podapolipidae), that lives beneath the elytra of Chrysophtharta agricola (Chapuis) (Coleoptera: Chrysomelidae). In each of 2 yr, prevalence of mites on beetles began each spring at about 10–20% but gradually increased to 80–100% by late summer. Overlap of adult beetle generations at this time (i.e., the parental generation mating with the F1 generation) is essential for the persistence of these mites. Mites exhibited temporal change in their spatial distribution on beetles; these changes were probably a response to beetle activity (e.g., emergence from diapause) and the need for dispersal from parental to F1 generation beetles. Prevalence and mean intensity of mites was higher on female beetles compared with male beetles. Female bias of sexually transmitted infection has been predicted in animals but hitherto observed only in primates. We speculate that variable male mate-finding success is the cause of these sex-based differences of mite infections, and that female bias in sexually transmitted disease (STD) infection will be widespread in the animal kingdom.

Small differences have been reported in the internal transcribed spacer 1 (ITS1) region among strains of Neospora caninum. We compared ITS1 sequences among 6 N. caninum strains analyzed in our laboratory, including 2 strains that have not been examined previously (NC-Illinois and NC-Bahia). Five sequences showed 100% similarity and also were identical to 7 of 11 sequences that were previously reported by others. In contrast, initial attempts to sequence the ITS1 of NC-Bahia generated 12 nucleotide differences compared with the other 5 strains, and several ambiguous bases. However, the single band containing the ITS1 region, as observed after electrophoresis on a 2% agarose gel, became divided into 2 distinct bands when reanalyzed using 5 or 10% polyacrylamide gel electrophoresis (PAGE), and the ITS1 within these separate bands were sequenced without ambiguity. The other 5 N. caninum strains were also reexamined using PAGE, and in each strain 2 distinct bands were discovered. In comparison, 2 strains of Toxoplasma gondii continued to show only 1 band when examined using PAGE. The ITS1 sequence of NC-Bahia, from Brazil, differs in several base pairs from those of North American and European strains of N. caninum. Intrastrain variation of the ITS1 region appears to be common in N. caninum, in contrast to T. gondii.

Gangliosides were isolated from Trypanosoma brucei and analyzed by thin-layer chromatography (TLC) and TLC immunostaining test. Four species of gangliosides, designated as G-1, G-2, G-3, and G-4, were separated by TLC. G-1 ganglioside had the same TLC migration rate as GM3. In contrast, G-2, G-3, and G-4 gangliosides migrated a little slower than GM1, GD1a, and GD1b, respectively. To characterize the molecular species of gangliosides from T. brucei, G-1, G-2, G-3, and G-4 gangliosides were purified and analyzed by TLC immunostaining test with monoclonal antibodies against gangliosides. G-1 ganglioside showed the reactivity to the monoclonal antibody against ganglioside GM3. G-2 was recognized by the anti-GM1 monoclonal antibody. G-3 showed reaction with the monoclonal antibody to GD1a. G-4 had the reactivity to anti-GD1b monoclonal antibody. Using 4 kinds of monoclonal antibodies, we also studied the expression of GM3, GM1, GD1a, and GD1b in T. brucei parasites. GM3, GM1, GD1a, and GD1b were detected on the cell surface of T. brucei. These results suggest that G-1, G-2, G-3, and G-4 gangliosides are GM3 (NeuAcα2-3Galβ1-4Glcβ1-1Cer), GM1 (Galβ1-3GalNAcβ1-4[NeuAcα2-3]Galβ1-4Glcβ1-1Cer), GD1a (NeuAcα2-3Galβ1-3GalNAcβ1-4[NeuAcα2-3]Galβ1-4Glcβ1-1Cer), and GD1b (Galβ1-3GalNAcβ1-4[NeuAcα2-8NeuAcα2- 3]Galβ1-4Glcβ1-1Cer), respectively, and also that they are expressed on the cell surface of T. brucei.

Two new species of Cardicola (Digenea: Sanguinicolidae) are described from the heart of drums (Perciformes: Sciaenidae) in and off Mississippi and Louisiana. Cardicola palmeri n. sp. infects the black drum, Pogonias cromis, and, in mature individuals, is distinguished from its congeners by the combination of a body 1.9–2.9 times longer than wide, an esophagus 44–52% of the body length, posterior ceca 1.0–1.8 times longer than the anterior ones, a gland encircling the seminal vesicle that is 30–37% of maximum body width in diameter, and a sinistral and nearly medial male pore. Cardicola currani n. sp. infects the red drum, Sciaenops ocellatus, and is distinguished from its congeners by the combination of posterior ceca 1.8–4.2 times longer than anterior ones, a male pore located well posterior to the ootype, a female pore and transverse metraterm located just anterior to the level of the ootype, and a medial and posttesticular ovary located at the level of the terminal ends of the posterior ceca. No adult blood fluke had been described previously from the northern Gulf of Mexico west of Florida. Each of the 2 flukes infected only 1 host species despite the hosts being sympatric.

Parapharyngodon duniae n. sp. (Nematoda: Pharyngodonidae) in the large intestine of the veined tree frog, Phrynohyas venulosa, from the Area de Conservación Guanacaste in northwestern Costa Rica is described and illustrated. Parapharyngodon duniae n. sp. represents the 34th species assigned to the genus, the 10th species from the Neotropical Realm, and only the third species to parasitize anurans. It is distinguished from the other Neotropical species by having postbulbar ovaries and a prebulbar excretory pore.

Cosmocerca vrcibradici n. sp. and Oswaldocruzia vitti n. sp., intestinal parasites of Prionodactylus eigenmanni (type host) and P. oshaughnessyi, are described and illustrated. Of the 19 valid species of Cosmocerca, C. vrcibradici n. sp. represents the 12th Neotropical species and the first species to be reported from lizard hosts. Of the 76 valid species of Oswaldocruzia, O. vitti n. sp. represents the 14th Neotropical species and the eighth species to be reported from lizard hosts. In addition, 1 species of Digenea, Mesocoelium monas, and 1 species of Acanthocephala, Acanthocephalus saurius, were found.

The sequence of the Gyrodactylus salaris Malmberg, 1957, large subunit, or 28S, ribosomal RNA (rRNA) gene has been determined. This gene is the final portion of the Gyrodactylus rRNA gene operon to be sequenced and results in the first complete sequence of all rRNA genes and spacers from a monogenean. The nucleotide sequence was used to predict the secondary structure of the large subunit rRNA, and regions of conserved and variable sequence and structure were identified. The site where the 5′ terminus of the 5.8S rRNA binds to a region within the large subunit rRNA was predicted and complements the anticipated interaction of the 3′ terminus of the 5.8S with the 5′ terminus of the large subunit rRNA. The large subunit gene may be useful in phylogenetic analysis of the Monogenea or Platyhelminthes and comparisons with other eukaryotes. The variable domains C and H may be most suitable for this purpose.

The molecular mechanisms by which the malarial parasite has managed to develop resistance to many antimalarial drugs remain to be completely elucidated. Mutations in the pfmdr1 gene of Plasmodium falciparum, as well as an increase in pfmdr1 copy number, have been associated with resistance to the quinoline-containing antimalarial drugs. We investigated the mechanisms of drug resistance in Plasmodium using a collection of P. yoelii lines with different drug resistance profiles. The mdr1 gene of P. yoelii (pymdr1) was identified and characterized. A 2- to 3-fold increase in the pymdr1 gene copy number was observed in the P. yoelii ART line (artemisinin resistant) when compared with the NS parental line. The pymdr1 gene was mapped to a chromosome of 2.1 Mb in all lines analyzed. Reverse transcriptase–polymerase chain reaction and Western blot experiments confirmed the expression of the gene at the RNA and protein levels.

Various monoclonal antibodies (mAbs) against Toxoplasma gondii RH tachyzoites were used for flow cytometric detection of intracellular parasites in murine splenic lymphocytes. Tg110 and Tg563 (reacting with the major surface protein SAG1), Tg505 (with another surface protein SAG2), Tg695 and Tg786 (with rhoptry proteins), Tg507, Tg621, and Tg317 (with dense granule proteins), Tg536 (with a microneme protein), and Tg685 (with a cytosol antigen) were the mAbs used. After an in vitro infection of lymphocytes with tachyzoites and reactions with the different mAbs, flow cytometry was performed using an indirect immunofluorescent technique. The proportions of whole infected lymphocytes and of each infected lymphocyte phenotype, CD4 T cells, CD8 T cells, and B cells, were determined, and their fluorescent intensities were quantified. The best reaction was seen when Tg110 or Tg695 was used as the mAbs. The results suggest that mAbs against surface or rhoptry proteins are highly useful for the flow cytometric detection of intracellular T. gondii in host cells.

Cryptosporidium spp. has been found in more than 150 species of mammals, but there has been no report in mongooses. In this study, we report the isolation of Cryptosporidium sp. in a banded mongoose Mungos mungo, which was brought from Tanzania to Japan; the isolate was analyzed genetically to validate the occurrence of a new, host-adapted genotype. Cryptosporidium diagnostic fragments of 18S ribosomal RNA and 70-kDa heat shock protein genes were amplified from this isolate and compared with the other Cryptosporidium species and genotypes reported previously. Analyses showed that the mongoose isolate represents a new genotype, closely related to that of bears.

Until now, Dicrocoelium sp. eggs have only been recorded from European and 1 North American archaeological sites. We present evidence for the first record of Dicrocoelium sp. from an African archaeological site. A paleoparasitological study was conducted on 7 coprolite samples from K2, a Late Iron Age site on the farm Greefswald, in the Northern Province of South Africa. Standard parasitological analysis revealed the presence of Dicrocoelium sp. and Trichuris sp. eggs. Today, the parasite does not occur in this region. Trichurid eggs are a relatively common find in paleoparasitological analysis. The presence of Dicrocoelium sp. provides new clues about the antiquity of this parasite, as well as aspects of ancient environment, climate, and interactions among humans, animals, and parasites.

Twenty-two Florida black bear (Ursus americanus floridanus) cubs (≤12 mo old) from Florida were examined for endoparasites between 1998 and 2003. Eleven species of helminths were found (8 nematodes, 2 trematodes, and 1 acanthocephalan). The most prevalent helminths were Ancylostoma caninum (64%), Macracanthorhynchus ingens (36%), Strongyloides sp. (27%), Baylisascaris transfuga (23%), and A. tubaeforme (14%). New host records include A. tubaeforme and Brachylaima virginianum. This is the first report of B. transfuga in black bears from Florida.

The muscular anatomy and fine structure of the antennae of adult Anthosoma crassum are described from specimens collected from jaws of shortfin makos. Casual observations of live parasites showed that embedded adult A. crassum can move toward or away from the host by shortening or lengthening the antennae, respectively. These movements were controlled by antennal retractor muscles. Microprotuberances located on the distal antennal segments may increase friction between these copepods and their hosts, thus strengthening parasite attachment.

The pathogenicity of female Philometra obturans (Prenant, 1886) was investigated. Fish samples were taken in May and June 2001 from Lake Littoistenjärvi, in southwest Finland. The intensity of infection varied between 1 and 4 worms, these being found in the afferent artery of the 2 outermost gill arches. The prevalence was 33.9% (n = 59). Host condition and tissue reactions were investigated through dissection and analysis of histopathology. The parasites obstructed the gill arteries, and the arterial wall was stretched around intact worms. Hypertrophy of the arterial wall was observed, especially when only disintegrating remains of the parasites were present. Infected pike were in a poorer condition after catch, transportation, and holding in aquaria and thus less tolerant to stress than uninfected pike. This was correlated with the number of infected gill arteries (P < 0.05). The results show that infection with this nematode has a detrimental effect on the host when combined with environmental or physiological stress and may result in host mortality.

We evaluated the proportion of variants of circumsporozoite protein (cp) gene 26 and cp29 antigenic epitopes of Plasmodium falciparum infecting patients among 3 provinces in western Thailand, in addition to published variants from Gambia. The proportion of patients coinfected with cp26 and cp29 strains was significantly higher in patients reporting to malaria clinics in Tak than in Kanchanaburi and Ratchaburi and higher in Kanchanaburi than in Ratchaburi. In western Thailand, coinfection with cp26 and cp29 appears to increase with increasing latitude. There were also significant differences in proportion of these variants among Thai provinces and Gambia. An association of patient human leukocyte antigen (HLA) class II genotype was associated with P. falciparum strains. There were significant associations among the HLA-DQA alleles in patients, the province of origin, and cp variants of P. falciparum.

Ergasilus megaceros (Copepoda: Ergasilidae) was recovered from the nasal fossae (lamellae) of the olfactory sac in 1 (1.8%) of 56 sea lampreys, Petromyzon marinus Linne, 1758, collected in May 2002 from the Cheboygan River, Michigan. Although the sea lamprey is a new host record for E. megaceros, this fish species may not be a preferred host because of its low prevalence. Ergasilus megaceros is the second ergasilid species reported from the sea lamprey in North America. This is the third report of an ergasilid species infecting the nasal fossae of fishes in North America, with E. rhinos being the only other species reported from this site.

Spirurid nematodes of the genus Thelazia cause ocular infection in several mammals and are transmitted by dipteran flies. Of the 10 described species, T. gulosa, T. rhodesi, T. skrjabini (affecting cattle), T. lacrymalis (affecting horses), and T. callipaeda (affecting carnivores and humans) are the most common. The aim of this study was to characterize the first internal transcribed spacer (ITS1) ribosomal DNA sequences of these 5 species. The length of the ITS1 sequences ranged from 357 bp (T. lacrymalis) to 905 bp (T. callipaeda). Interspecific differences ranged from 35 to 77%. Polymorphic sites were detected in each species. Intraspecific variation varied from 0.3 to 2.5%. The characteristics of the ITS1 of Thelazia spp. show similarities to those of other Spirurida. The findings of this present study show that the ITS1 represent a powerful genetic marker for the molecular identification of eyeworms and a useful tool for developing molecular epidemiological techniques to study Thelazia spp. transmission patterns and prevalence in definitive and intermediate hosts.

We describe the first account of the nematode Gyrinicola batrachiensis in the gastrointestinal tracts of tadpoles of Bufo terrestris, Gastrophryne carolinensis, Hyla femoralis, H. squirella, Rana heckscheri, and R. utricularia. Infection by G. batrachiensis was also noted in tadpoles of R. catesbeiana and R. clamitans, species previously described as harboring these nematodes. These observations represent a major expansion of the known geographical range of G. batrachiensis to include the southeastern United States. In our study, female G. batrachiensis nematodes in ranid host species were didelphic, but females in B. terrestris tadpoles were monodelphic. Furthermore, only female nematodes were found in the latter host species. Such variations in nematode uterine morphology and sex ratios among these tadpole host species are in accordance with the apomictic and haplodiploid reproductive strategies previously reported for G. batrachiensis. These observations also support a generalized pattern relating G. batrachiensis reproductive modes and the life cycles of their tadpole hosts.

Recently, several polymerase chain reaction (PCR)–based methods for detection and genetic identification of haemosporidian parasites in avian blood have been developed. Most of these have considerably higher sensitivity compared with traditional microscope-based examinations of blood smears. These new methods have already had a strong impact on several aspects of research on avian blood parasites. In this study, we present a new nested PCR approach, building on a previously published PCR method, which has significantly improved performance. We compare the new method with some existing assays and show, by sequence-based data, that the higher detection rate is mainly due to superior detection of Plasmodium spp. infections, which often are of low intensity and, therefore, hard to detect with other methods.

The effects of Haemoproteus belopolskyi, Haemoproteus fringillae, and Haemoproteus lanii on the survival of their vector the biting midge Culicoides impunctatus were examined. Wild-caught females were infected experimentally by allowing them to feed on naturally infected birds. A group of flies, which fed on an uninfected bird, was used as a control. There was a highly significant difference in the survival rate between the noninfected group and all groups of infected flies for postfeeding (PF) periods 1–2, 3–4, 5–6, and 7–8 days. For days 7–8, the percentage of noninfected surviving flies was 4.4 times higher than the percentage of infected surviving flies, testifying to the detrimental effects of avian Haemoproteus infections on the longevity of C. impunctatus. The mortality rates of the infected flies were highest for days 1–2 and 3–4 PF, indicating possible negative effects of ookinetes and early oocysts on the vector.

Haemoproteus belopolskyi of blackcap, Sylvia atricapilla, underwent sporogony in wild-caught female biting midges, Culicoides impunctatus, which were experimentally infected by feeding them on naturally infected birds. The engorged flies were held for 8–12 days to allow development of sporozoites and then aspirated and triturated in 0.85% saline. Seven uninfected nestlings of blackcap at the age of 20–21 days were inoculated into the pectoral muscle with 0.3 ml of the slurry containing approximately 45 sporozoites. Parasitemia of H. belopolskyi developed in 6 nestlings, with a prepatent period of 11– 12 days. The maximum parasitemia varied between 0.9 and 16% of erythrocytes in different experimental hosts. Culicoides impunctatus is an experimental vector of H. belopolskyi. It is likely to be the important natural vector of Haemoproteus spp. of passerine birds in Europe.

The effect of time on genetic differentiation was studied among infrapopulations of mature specimens of the hemiurid fluke, Lecithochirium fusiforme, a parasite of marine fishes. Genetic distances and genetic structure within and among different temporal samples of a geographical population were investigated using starch gel electrophoresis, by screening 6 polymorphic loci in 2 groups of infrapopulations corresponding to different sampling data, i.e., winter 1997–1998 and autumn 1998. The genetic distance among infrapopulations was low (D = 0.000–0.058 ± 0.041). However, genetic divergence among infrapopulations from the same geographic location was clearly lower within each temporal sample (G_ST = 0.021 and 0.034) than the corresponding value obtained for 12 infrapopulations sampled at different seasons of the year (G_ST = 0.067). These results suggest the existence of a relatively important temporal effect that accounts for the differences in genetic variability among adult infrapopulations of L. fusiforme. Therefore, a hypothetical temporal gene flow favored by the existence of persistent life-cycle stages of this species in paratenic hosts is not sufficient to mask the temporal differentiation caused by genetic drift.

Two cases of submandibular echinococcosis have been reported in adult residents of India (Kini et al., 1997; Sahni et al., 2000). It was stated in both publications that the cestode was E. oligarthrus, “a rare variety of E. granulosus”, based on morphogical characteristics. It was also mentioned that 3 cases of E. granulosus with similar submandibular metacestode had been previously diagnosed in India. The metacestode is readily distinguishable from that of E. oligarthrus. The final hosts of E. oligarthrus are several neotropical species of wild cats and metacestode occurs in hystricognath, all of them absent in India. The larval stage of all 4 species of Echinococcus can be differentiated on the basis of the form and dimensions of rostellar hooks of protoescoleces. With the information provided in the two papers we concluded that the 2 cases of echinococcosis described in India were due to E. granulosus.