The influence of lectins on Cryptosporidium parvum oocyst agglutination and on attachment to both fixed Madin Darby Canine Kidney (MDCK) cells and fixed HCT-8 (human colorectal epithelial) cells was examined. Oocyst cell wall characteristics were examined by transmission electron microscopy. Lectin-free oocysts were shown to adhere equally to both MDCK cells and HCT-8 cells. In MDCK cells, the addition of 1–25 μg/ml Codium fragile lectin, 10 μg/ml Maclura pomifera lectin, 10 μg/ml Helix pomatia lectin, and 10–200 μg/ml Artocarpus integrifolia lectin significantly increased attachment to at least 1 of the cell cultures as compared to oocysts incubated without any lectin. The lectin-enhanced attachment was reversed by co-incubation of lectin treated-oocysts with 250 mM of each specific sugar (for a given lectin). In agglutination assays, concentrations as low as 0.5 μg/ml of C. fragile, M. pomifera, and A. integrifolia lectin agglutinated oocysts within 60 min. Finally, in TEM samples, colloidal gold conjugated-lectins from A. integrifolia, C. fragile, H. pomatia, and M. pomifera attached to oocysts, and this could be competitively inhibited by a lectin-specific sugar. This suggests that C. parvum oocysts are highly reactive to N-acetyl galactosamine-binding lectins and that the presence of N-acetyl-galactosamine containing molecules on oocysts can potentially help in oocyst attachment to host cells.

During 4 consecutive collecting seasons from 1998 through 2001, 77,326 common beach snails (Stagnicola emarginata, Lymnaeidae) were examined for infections by Trichobilharzia stagnicolae from multiple sites on Walloon Lake, Higgins Lake, and Lake Leelanau, located in the northern region of the lower peninsula of Michigan. Snails were examined for infections using the light-box technique (exposure to bright fluorescent light). The prevalence of infected snails varied significantly among lakes within a year, between years in a lake, at a site from year to year, and at a site over a collecting season. Overall annual prevalence ranged from 0.54% (1999) to 1.32% (2001) on Walloon Lake, from 0.56% (2001) to 1.06% (1998) on Higgins Lake, and 0.30% (2001) to 0.89% (2000) on Lake Leelanau. Generally, the peak in prevalence coincided in July on the lakes. Prevalence was found to increase with snail length in all lakes. A comparison of the light-box technique and crushing snails demonstrated that prevalence varied in magnitude by technique as much as 1.2 and 5.7 times.

Using a range of parameters, the ability of rats (Rattus norvegicus) to successfully transmit Echinostoma friedi to the next host was examined under experimental conditions. The concept of Experimental Transmission Success (T_M), defined as the number of hosts that become successfully infected after exposure to a number of infective stages produced by a previous host per unit of inoculation at which this latter host was exposed, was introduced. Using data for the egg output and miracidium hatching and infectivity, the T_M permits us to estimate the ability of a particular defintive host species to successfully transmit a parasite species. This concept may be also useful to compare the transmission fitness of a parasite in different definitive host species. Moreover, variations of the Experimental Transmission Success over the course of the infection were calculated by the use of the Weekly Experimental Transmission Success (T_MW). Overall, considering the complete duration of the experiment, the T_M of E. friedi using rats as definitive hosts was 0.68 infected snails/metacercaria. However, positive values of the T_MW were only obtained from 2 to 4 wk post-infection, with a maximum during the third wk post-infection. When comparing the T_M values of E. friedi in rats with those calculated in hamsters on the basis of previously published data, E. friedi appears to be more appropriate to move through this portion of its life cycle when using hamsters (Mesocricetus auratus) as the final host than rats.

Monocotyle guttatae n. sp. is described from the gills of the ray Dasyatis guttata (Bloch and Schneider, 1801) (Dasyatidae) off the coast of Brazil The new species can be readily differentiated from the other species of the genus in having a male copulatory organ with 2 loops and an accessory piece, 5–7 sclerites on the marginal haptoral papillae, and the absence of accessory sclerites on the dorsal surface of the posterior body. The present record confirms the presence of the genus in the subtropical waters of South America.

The host specificity of some parasites can be reinforced by morphological specialization for attachment to mobile hosts. For example, ectoparasites with adaptations for attaching to hosts of a particular size might not be able to remain attached to larger or smaller hosts. This hypothesis is suggested by the positive correlation documented between the body sizes of many parasites and their hosts. We adopted an ecomorphological approach to test the attachment hypothesis. We tested the ability of host-specific feather lice (Phthiraptera: Ischnocera) to attach to 6 novel species of pigeons and doves that vary in size by nearly 2 orders of magnitude. Surprisingly, Rock Pigeon lice (Columbicola columbae) remained attached equally well to all 6 novel host species. We tested the relative importance of 3 factors that could facilitate louse attachment: whole-body insertion, tarsal claw use, and mandible use. Insertion, per se, was not necessary for attachment. However, insertion on coarse feathers of large hosts allowed lice to access feather barbules with their mandibles. Mandible use was a key component of attachment regardless of feather size. Attachment constraints do not appear to reinforce host specificity in this system.

In the present study, PCR has been applied to detect and analyze DNA of Babesia spp. extracted from Ixodes ricinus ticks. Collection of I. ricinus was made in 6 forested areas of Zachodniopomorskie Voivodship, Poland, during 2 seasonal peaks of tick activity, i.e., spring and autumn, 2001. In total, 1,328 I. ricinus were collected and processed for PCR with F34 and R323 primers. Babesia spp. was detected in 28 (2% of 1,328 tested) ticks; 26 were identified as B. divergens. The other 2 were identified as B. microti. PCR was conducted with 18S rRNA specific primers and sequencing was processed to precisely identify and compare these isolates with B. microti and B. divergens sequences from Europe, North America, and Asia obtained from the GenBank. Analysis revealed that sequences of B. microti from northwestern Poland are almost identical (99.94%) with those referred to as “Munich strain”; both form a clade different from other European strains, as well as those from Asia and North America (called B. microti, sensu stricto). An investigation performed with B. divergens sequences showed that the sequence from northwestern Poland is 99.94% homologous to an isolate from Ireland (“Purnel”), and differs in just a few nucleotides from other European sequences. Phylogenetic analysis revealed that the sequence of B divergens isolated from Polish ticks form a group that comprise 4 European sequences from Great Britain and Ireland and is, therefore, closely related to other European and North American B. divergnens sequences.

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 50 free-range chickens (Gallus domesticus) from Amazon, Brazil, was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT) and found in 33 (66%) chickens with titers of 1:5 in 3, 1:10 in 2, 1:20 in 1, 1:40 in 1, 1:80 in 2, 1:160 in 5, 1:200 in 9, 1:400 in 5, 1:800 in 2, 1:1,600 in 2, and 1:3,200 or higher in 1. Hearts and brains of 33 seropositive chickens were bioassayed individually in mice. Tissues from 17 seronegative chickens were pooled and fed to 2 T. gondii–free cats. Feces of cats were examined for oocysts, but none was found. Toxoplasma gondii was isolated from 24 chickens with MAT titers of 1:5 or higher. Genotyping of these 24 T. gondii isolates by polymorphisms at the SAG2 locus indicated that 14 were type I, and 10 were type III; the absence of type II strains from Brazil was confirmed. Fifty percent of the infected mice died of toxoplasmosis, irrespective of the genotype.

In an attempt to study the occurrence of concomitant immunity in filarial infections, jirds (Meriones unguiculatus) were experimentally infected with Acanthocheilonema viteae, and patent animals were superinfected with a defined dose of A. viteae stage 3 larvae (L3). Infected animals harbored significantly less worms deriving from the superinfection than the control group (P < 0.05, 56.2%, and 63.4% protection), as shown by analysis of female worms 6 wk after superinfection on the basis of their developmental status and their length. This protection was not due to contact with L3 antigens because a significant reduction of worm burdens deriving of a superinfection was also observed after subcutaneous implantation of a single female worm (P < 0.05, 40.2% and 64.9% protection). The induced protective responses target L3 and restrict their migration because an established infection resulted in a reduction of L3 recovery (95.6% and 94.3%, P < 0.001) from tissues of jirds at day 5 after superinfection. Other data show that L3 from a superinfection are trapped within eosinophil-rich granulomas, which is likely to create unfavorable conditions for the worms and to lead to later death. Taken together, established A. viteae-infections partially protect hosts against homologous superinfection by an immune-mediated mechanism and, thus, regulate the population density of the parasites within the host by concomitant immunity.

Infection with mosquito-born filarial nematodes occurs when hosts are bitten by a vector carrying the infective third stage larvae (L3) of the parasites. These larvae, deposited on the skin by the feeding mosquito, are presumed to enter the skin via the vector-induced puncture wound. Larvae of Brugia spp. must then migrate from the entry site, penetrate various skin layers, and locate a lymphatic vessel that leads to their lymphatic predilection site. We have recently established an intradermal (ID) infection model using B. pahangi and the Mongolian gerbil, allowing us to investigate the migratory capability of B. pahangi. Larval and adult parasites recovered from the peritoneal cavities of gerbils were capable of establishing an infection following ID (larvae) or subcutaneous (adult) injection. Third and fourth stage larvae both migrated away from the injection site within hours, although data suggest they localize to different lymphatic tissues at 3 days postinfection (DPI). Immature adult (28 day) B. pahangi also migrated away from their SC inoculation site within 7 DPI. Mature (45 day) adult B. pahangi displayed little migration away from the SC infection site, suggesting tissue migration may be limited to developing stages of the parasite.

Membership and richness of infracommunities and component communities of myxozoan fauna of the banded killifish (Fundulus diaphanus) from freshwater localities in Ontario, Quebec, New York State, New Brunswick, Nova Scotia, and Maryland were studied. Five species of parasites were found: Myxobolus diaphanus (Fantham, Porter, and Richardson, 1940) (connective tissue throughout the body and head), Myxobolus funduli (Kudo, 1918) (interlamellar), Myxobolus neurophilus (Guilford, 1963) (optic tectum of the brain), Myxobolus sp. (connective tissue, typically adjacent to vertebrae), and Sphaerospora sp. (kidney tubules). The most abundant species locally and regionally was M. diaphanus, occurring at prevalences of 14.2 to 93.3% at 6 of 9 localities. Myxobolus funduli and Myxobolus sp. were at 3 and 2 localities respectively, while M. neurophilus and Sphaerospora each occurred at single localities. Four of the 5 myxozoans appear to be specific to fundulids, the exception being M. neurophilus, which is typically a parasite of Perca flavescens. Mean infracommunity richness was 0–1.2. Component community richness was 0–3 species. The fauna is similar in composition to that described from the spottail shiner (Notropis hudsonius) in the Great Lakes in being dominated by histozoic myxobolids and in having maximum prevalence at any single locality correlate positively with geographical distribution. Moreover, mean infracommunity richness was correlated with percentage of hosts infected with any species at a locality, and maximum infracommunity richness was correlated with component community richness. Probably because fewer species of myxozoans of fundulids occur in the regional pool, myxozoan communities encountered in the present study are generally less rich than those described from N. hudsonius. It appears that dispersal of relatively resilient myxospores through such a mechanism as piscivory effectively distributes these parasites over the landscape, while the more delicate actinospores serve to ensure colonization by amplifying species' prevalence at a specific locality and thereby contributing to initial establishment. As such, these types of myxozoans, though they are autogenic, having their entire life cycle normally completed within the aquatic environment, behave more like allogenic parasites that rely on birds and mammals as definitive hosts.

Cryptosporidium parvum has become the focus of numerous studies on waterborne disease and transmission in response to outbreaks endangering populations worldwide. The Foci Detection Method–Most Probable Number Assay (FDM– MPN) is an in vitro cell culture method that has been developed and used to determine the quantity of infectious C. parvum oocysts. This research evaluated 2 vendor's producing oocysts, Sterling Parasitology Laboratory (SPL) and Pleasant Hill Farms (PHF) (now known as Bunch Grass Farms as of 12/03), classified as young (<30 days) and aged (>165 days), for comparison of treatments (bleach, antibiotic, no treatment) before cell culture, as well as an age study, to determine any lot-to-lot differences and vendor differences regarding the rate of decline in infectivity. Bleach treatment (0.525%) appeared to be the optimum method for the FDM–MPN with regards to maximum infectivity, efficient disinfection, with no visible antagonistic affects on the C. parvum oocysts. The age study revealed that lot-to-lot variability within each vendor stayed within 1 log₁₀ difference, while the rates of decline in infectivity measured until 107 and 120 days of age when stored at 4 C for SPL and PHF were −0.016 and −0.014 log₁₀ infectious oocysts/day, respectively. These results provide insight regarding C. parvum oocyst viability in a fecal population, as well as useful knowledge for further methods development.

Strongyloides procyonis Little, 1966 was detected about 45 years ago in raccoons (Procyon lotor) of southern Louisiana, U.S.A., and was demonstrated experimentally to cause creeping eruption and a short-lived intestinal infection in a healthy human volunteer. After its description and demonstration of its pathogenicity in humans, S. procyonis has not been found in raccoons in North America despite repeated surveys. During a survey on feral raccoons in Japan, S. procyonis parasitic females were identified in 66 (28.3%) of 233 raccoons collected between May 2004 and January 2005. The number of parasitic females recovered from individual raccoons was 1–197 (geomean, 3.2). Both the morphological features and the nucleotide sequences of the small and large subunit ribosomal RNA genes (SSU/LSU rDNA) of S. procyonis closely resembled those of zoonotic Strongyloides stercoralis. The sequences of internal transcribed spacer (ITS)1 and 28S rDNA could differentiate clearly these 2 species. Awareness of S. procyonis in raccoons in North America and other places worldwide where raccoons are introduced and naturalized is important to assess the epidemiological significance of this potentially zoonotic helminth species.

Parasitological examination of feces from 44 Emys orbicularis from Galicia (NW Spain) revealed the presence of 2 new eimerian species, Eimeria gallaeciaensis sp. n. and E. emydis sp. n., as well as E. mitraria (Laveran and Mesnil, 1902) Doflein, 1909. Oocysts of E. gallaeciaensis n. sp. were found in 20 of 44 (45.4%) turtles and are subspherical to lightly ovoid– ellipsoid, 19.3 × 16.0 (17–22 × 15–18) μm, shape index 1.2 (1.1–1.3), with a smooth, single-layered wall. Micropyle and polar granule are absent, but an oocyst residuum is present. Sporocysts are ellipsoid, 9.7 × 5.1 (9–10 × 5–6) μm, shape index 1.9 (1.7–2.0), each with a sporocyst residuum and a conical Stieda body usually bearing 1–4 short and thin projections. Oocysts of E. emydis n. sp. were found in the feces of 5 of 44 (11.4%) turtles and are ovoid, rarely pear-shaped, 22.6 × 17.0 (20–25 × 15.5–18) μm, shape index 1.3 (1.2–1.5), with a smooth, single-layered wall with a slight thinning at the pointed end. Micropyle and polar granule are absent, and an oocyst residuum is present. Sporocysts are ellipsoid, 11.4 × 6.0 (9–13 × 5–7) μm, shape index 1.9 (1.6–2.2), each with sporocyst residuum and a prominent Stieda body bearing 3–5 club-shaped projections. In addition to the new species described, this is the first report of E. mitraria parasitizing E. orbicularis.

Karyakartia egyptensis n. sp. (Digenea: Lepocreadiidae: Lepocreadiinae) is described from the intestine of the Jarbua terapon, Terapon jarbua (Forsskål), collected from the Red Sea off the coast of Hurghada, Egypt. Karyakartia egyptensis n. sp. differs from K. pambanense in that the former species possesses larger spiniform structures around the perimeter of the mouth (48–54 μm as compared with 25–30 μm), a uroproct rather than ceca that end blindly or form a cyclocoel, and a somewhat smaller egg (60–68 μm as compared with 70–73 μm).

Revealing diversity among extant blood flukes, and the patterns of relationships among them, has been hindered by the difficulty of determining if specimens described from different life cycle stages, hosts, geographic localities, and times represent the same or different species. Persistent collection of all available life cycle stages and provision of exact collection localities, host identification, reference DNA sequences for the parasite, and voucher specimens eventually will provide the framework needed to piece together individual life cycles and facilitate reconciliation with classical taxonomic descriptions, including those based on single life cycle stages. It also provides a means to document unique or rare species that might only ever be recovered from a single life cycle stage. With an emphasis on the value of new information from field collections of any available life cycle stages, here we provide data for several blood fluke cercariae from freshwater snails from Kenya, Uganda, and Australia. Similar data are provided for adult worms of Macrobilharzia macrobilharzia and miracidia of Bivitellobilharzia nairi. Some schistosome and sanguinicolid cercariae that we recovered have peculiar morphological features, and our phylogenetic analyses (18S and 28S rDNA and mtDNA CO1) suggest that 2 of the new schistosome specimens likely represent previously unknown lineages. Our results also provide new insights into 2 of the 4 remaining schistosome genera yet to be extensively characterized with respect to their position in molecular phylogenies, Macrobilharzia and Bivitellobilharzia. The accessibility of each life cycle stage is likely to vary dramatically from one parasite species to the next, and our examples validate the potential usefulness of information gleaned from even one such stage, whatever it might be.

Malmiana buthi n. sp. is described from the body and fins of the fluffy sculpin, Oligocottus snyderi, the tidepool sculpin, Oligocottus maculosus, and the woolly sculpin, Clinocottus analis, collected from tidepools at Horseshoe Cove on the Bodega Marine Reserve in Sonoma County, California. Prevalence of the leech was 32.6% on live-caught O. snyderi; mean intensity on O. snyderi was 3.3 leeches per fish, with a range from 1 to 7. The leech is not known to exceed 8 mm total length. The body is smooth, lacking papillae, tubercles, or pulsatile vesicles. Two pairs of crescentiform eyes are present on the oral sucker, and 1 pair of punctiform eyes occurs on the second annulus of the trachelosome. The caudal sucker has 14 small punctiform ocelli spaced evenly around the margin. The last 9 segments of the urosome have pairs of large punctiform ocelli both dorsally and ventrally. Body and caudal sucker pigmentation is uniformly reddish brown dorsally and ventrally with segmental, lateral, unpigmented areas on both the urosome and trachelosome; pigmentation on the oral sucker is in the form of a cross. There are 5 pairs of testisacs; accessory gland cells on the atrial cornua and vector tissue are absent.

Molecular techniques were used to examine the phylogenetic relationships among Hepatozoon species isolated from 13 foxes and 15 opossums from Brazil, and from 15 dogs, 20 foxes, 45 rodents, and 330 domestic cats from Spain. Hemogregarine infection was confirmed by amplification of the 18S rRNA gene and later sequencing. No hemogregarine infections were found in opossums. The prevalence of Hepatozoon in canids ranged from 26.6% (symptomatic domestic dogs) to 90% (Spanish foxes). Four different H. canis genotypes were detected, as well as an H. americanum-related protozoan (97% identical to the USA strain). Two Spanish cats were parasitized by a Hepatozoon species (0.6% prevalence) that showed 96% sequence identity to H. canis. DNA amplification assays performed on Spanish rodents showed 2 bank voles (Clethrionomys glareolus) to be infected by a Hepatozoon species (4.44% prevalence) with 95% sequence identity to Hepatozoon sp. from cats. Phylogenetic analysis showed Hepatozoon to be a monophyletic genus, in which species from carnivorous mammals (Hepatozoon sp. from cats, H. americanum and H. canis) appear as a sister lineage of that of lower vertebrates and rodents. This association suggests that H. americanum evolved in ticks and carnivores (either canids, or felids, or both) rather than in other ectoparasites and other types of mammal.

A new species of Trichuris is described. Trichuris pardinasi n. sp. was recovered from Phyllotis xanthopygus Waterhouse (Rodentia: Muridae: Phyllotini) in Sierra de la Ventana, Buenos Aires Province, and Pampa de Achala, Córdoba Province (Argentina). This is the first record of Trichuris parasitizing Phyllotini rodents. The new species can be differentiated from the other 10 species parasitizing rodents from South America by the absence of the spicular tube, spicular sheath with spines uniformly distributed, the length of spicule, the J-shaped proximal cloacal tube, and the nonprotrusive vulva. Also, a description of the bacillary band is provided. The present and the future findings of shared parasite fauna from both populations of P. xanthopygus in these disjunct areas will support the hypothesis of a continuous distribution of this host species at a past time.

Five new species of Acanthobothrium (Tetraphyllidea: Onchobothriidae) are described from the spiral intestine of the Freshwater whipray, Himantura chaophraya, in the Kinabatangan River in Malaysian Borneo. Based on criteria set forth in a previous categorization scheme for species of Acanthobothrium, these consist of 3 Category 1 species, Acanthobothrium asnihae n. sp., Acanthobothrium saliki n. sp., and Acanthobothrium zainali n. sp.; a Category 8 species, Acanthobothrium etini n. sp.; and a Category 2 species, Acanthobothrium masnihae n. sp.. Acanthobothrium asnihae n. sp. differs from all Category 1 species in its possession of a horizontal band of weak musculature that divides the posterior loculus in half. Among Category 1 species, A. saliki n. sp. differs from all but Acanthobothrium southwelli in its possession of postovarian testes. It differs from A. southwelli in its possession of fewer testes and a greater number of proglottids. Acanthobothrium zainali n. sp. differs from the 25 other Category 1 species in a combination of overall size, muscular pad and hook shape, arrangement and number of testes, ovary configuration in cross section, position of ovarian isthmus, and genital pore position. Acanthobothrium etini n. sp. is distinguished from all 5 other Category 8 species in its lack of testes from the proglottid antiporal and postporal regions and in testis number. Acanthobothrium masnihae n. sp. differs from the 35 other Category 2 species in its possession of fewer testes, postporal testes, or a greater number of proglottids. A key to Acanthobothrium species parasitizing H. chayophraya is presented. This represents the first report of Acanthobothrium from freshwater stingrays belonging to a family other than the Potamotrygonidae.

Australicola pectinatus n. gen., n. sp. (Pseudophyllidea: Triaenophoridae) is proposed to accommodate a new cestode from a deep-sea fish, the splendid alfonsino, Beryx splendens Lowe, 1834 (Beryciformes: Berycidae), from the Pacific coast of Tasmania. The new genus is placed in the Triaenophoridae, because it possesses a ventral uterine pore, marginal genital pore, and follicular vitellarium. Australicola is characterized by possessing a massive strobila with very short and wide, markedly craspedote proglottids; vitelline follicles forming a transverse equatorial band; a very deep and narrow genital atrium; a wide, convoluted vaginal canal; and unoperculate eggs. Australicola most closely resembles Eubothrium Nybelin, 1922 and Probothriocephalus Campbell, 1979 in having an unarmed scolex, an unarmed cirrus, the vagina anterior to the cirrus-sac, and cortical vitellaria. It differs from these 2 genera, in addition to the characteristics listed above, in possessing a dendritic rather than an entire ovary. Australicola pectinatus n. sp. is the third cestode described from B. splendens.

A new species of parasitic nematode, Procamallanus (Procamallanus) pacificus n. sp., is described from the stomach of the Pacific shortfinned eel, Anguilla obscura (type host), and from the speckled longfin eel, Anguilla reinhardtii, from northern New Caledonia (Melanesia, South Pacific); from Anguilla sp. (cf. obscura) from the Fiji Islands (Melanesia, South Pacific); and from the giant mottled eel Anguilla marmorata from Futuna Island (Wallis and Futuna Islands, Polynesia). Although a total of 450 nematodes were collected, all specimens were females; this suggests either an extremely rare occurrence of males or parthenogenetic reproduction in this species. Procamallanus pacificus differs markedly from all congeners from fish hosts in possessing a greater number (4–9) of caudal mucrons in the female and by other morphological features. This parasite might become a serious pathogen of cultured eels in the region of the South Pacific. Batrachocamallanus Jackson and Tinsley, 1995 is considered a junior synonym of Procamallanus Baylis, 1923, to which 2 species are transferred as Procamallanus occidentalis (Jackson and Tinsley, 1995) n. comb. and Procamallanus siluranae (Jackson and Tinsley, 1995) n. comb. One third-stage larva of Procamallanus (Spirocamallanus) sp. was also recorded from Anguilla sp. (cf. obscura) from the Fiji Islands.

Nematodes (1 male and numerous females) of the Philometridae were collected from the mesentery of 2 species of pimelodid catfishes, Calophysus macropterus and Perrunichthys perruno, from the Amazon River basin (fishmarket in Iquitos, Loreto District) in Peru. A detailed study of their morphology (including scanning electron microscopy) and a reexamination of the type and voucher specimens of Philometra amazonica Travassos, 1960, from Brazilian catfishes confirmed that they belong to this species and that Philometra (Alinema) alii Rasheed, 1963 is its junior synonym. Because of some marked morphological peculiarities of this species (presence of minute peribuccal sclerotized formations, a functional vagina and vulva in gravid female, and structure of the male tail), the validity of an independent genus, Alinema Rasheed, 1963, is confirmed, to which this species is transferred as Alinema amazonicum (Travassos, 1960) n. comb. This is the first record of this parasite from Peru, and P. perruno represents its new host record.

Two new species of Paraorygmatobothrium Ruhnke, 1994, P. janineae n. sp. and P. kirstenae n. sp., are described from the spiral intestine of 2 shark species of the Family Hemigaleidae: Hemigaleus microstoma and Hemipristis elongata. The 2 new cestode species differ from other members of Paraorygmatobothrium in vitelline follicle distribution and possession of a cephalic peduncle. The 2 new species differ from 1 another in total length, maximum width, scolex size, number of proglottids per strobila, and number of testes per proglottid. The generic diagnosis of Paraorygmatobothrium is emended to include the new species. The results of this study extend the distribution of Paraorygmatobothrium to include the carcharhinid shark family Hemigaleidae.

In total, 17 specimens of Conger orbignianus Valenciennes, 1847 from waters off Mar del Plata, Argentina (38°08′S, 57°32′W) were examined for parasitic nematodes. A new nematode species, Cucullanus pedroi n. sp., is described (prevalence 76.5%, x̄ intensity ± SD = 3.8 ± 2.7). The new species closely resembles some species parasitizing other anguilliform fishes; however, it can be distinguished from most of its congeners by the distribution pattern of caudal papillae (particularly fourth and eight pairs) and phasmids. Those congeners with similar pattern of papillae differ from the new species by the length of the spicules and gubernaculum and by the position of the excretory pore and deirids.

The genetic relationships among 9 taxa of Anisakis Dujardin, 1845 (A. simplex (sensu stricto), A. pegreffii, A. simplex C., A. typica, A. ziphidarum, A. physeteris, A. brevispiculata, A. paggiae, and Anisakis sp.) were inferred from sequence analysis (629 bp) of the mitochondrial cox2 gene. Genetic divergence among the considered taxa, estimated by p-distance, ranged from p = 0.055, between sibling species of the A. simplex complex, to p = 0.12, between morphologically differentiated species, i.e., A. ziphidarum and A. typica. The highest level was detected when comparing A. physeteris, A. brevispiculata, and A. paggiae versus A. simplex complex (on average p = 0.13) or versus A. typica (on average p = 0.14). Sequence data from the newly identified Anisakis sp. poorly aligned with other Anisakis species but was most similar to A. ziphidarum (p = 0.08). Phylogenetic analyses based upon Parsimony and Bayesian Inference, as well as phenetic analysis based upon Neighbor-Joining p-distance values, generated similar tree topologies, each well supported at major nodes. All analyses delineated two main claides, the first encompassing A. physeteris, A. brevispiculata, and A. paggiae as a sister group to all the remaining species, and the second comprising the species of the A. simplex complex (A. simplex (s.s.), A. pegreffii and A. simplex C), A. typica, A. ziphidarum, and Anisakis sp. In general, mtDNA-based tree topologies showed high congruence with those generated from nuclear data sets (19 enzyme-loci) and with morphological data delineating adult and larval stages of the Anisakis spp.; however, precise positioning of A. typica and A. ziphidarum remain poorly resolved, though they consistently clustered in the same clade as Anisakis sp. and the A. simplex complex. Comparison of anisakid data with those currently available for their cetacean-definitive hosts suggests parallelism between host and parasite phylogenetic tree topologies.

Fischthal and Nasir (1974) reported Neohaematotrephus brasilianum (as Cyclocoelum brasilianum) in the spotted-sandpiper Actitis macularia from Venezuela. Three voucher specimens from that report, deposited in the United States National Parasite Collection, however, differ from N. brasilianum by having the cirrus sac on the sinistral side of the body, which resembles N. facioi, N. arayae, and N. gendrei. The new species is similar to N. brasilianum by having vitelline follicles extending well anterior to the intestinal bifurcation and by having a short and laterally displaced cirrus sac whose posterior end does not reach the intestinal bifurcation, whereas all other members of Neohaematotrephus have a cirrus sac that is medially oriented. Neohaematotrephus gendrei and N. facioi have cirrus sacs that extend to the level of the intestinal bifurcation, and N. arayae has a cirrus sac that extends well posterior to the posterior margin of the ceca. By having the ovary on the sinistral side of the body, the new species is similar to N. brasilianum, N. gendrei, and N. arayae but differs from N. facioi, in which the ovary is dextral.

Iron chelating agents, which permeate through erythrocytic and parasite membranes, are effective against Plasmodium falciparum in vitro. However, the protective effect in humans is transient. We examined the antiplasmodial capacity of several iron chelators in vitro and in vivo. The chelators 3/3hb/2m and 3/2hb/b (together, MoB) were more effective against P. falciparum in vitro than desferrioxamine (DFO) and Salicylaldehyde isonicotinoyl hydrazone (SIH) (together, DoS). Despite similar pharmacokinetics of all iron chelators, mice infected with Plasmodium vinckei and treated with MoB succumbed to malaria, whereas DoS-treated mice survived. However, even in the surviving mice, peak parasitemias were above 30%. These results indicate that the direct effects of the drugs on the parasites were not responsible alone for the complete recovery of the mice. We suggest that the recovery is related to differential effects of the drugs on various immune functions. We concentrated on the effect of the iron chelators on B cell and T cell proliferation and on allogeneic stimulation (MLR), interleukin-10 (IL-10), γ-interferon (γ-IFN), tumor necrosis factor-α (TNF-α), and radical production. All the iron chelators examined inhibited the in vitro proliferation of B cells and T cells, and MLR. This may explain why iron chelators are only slightly efficient in treating human malaria. However, the inhibitory effects of MoB on B cell and T cell proliferation and on MLR were more pronounced than those of DoS. In addition, the release of free radicals by effector cells was inhibited to a greater extent by MoB than by DoS. These results may explain why MoB, which was more efficient in vitro, was not effective in vivo. The DoS effects on the in vitro secretion of cytokines correlate with their in vivo effect; there was a decrease of IL-10 and a parallel increase in γ-IFN and TNF-α production by human mononuclear cells. MoB, which could not rescue the animals from malaria, did not affect IL-10 and TNF-α, but reduced γ-IFN levels. Identical results were obtained when using monocytes instead of mononuclear cells (except for γ-IFN, which is not produced by monocytes). Our results indicate that an iron chelator, or any antiparasitic drug that kills the parasites in vitro, should also be selected for further evaluation on the basis of its reaction with immune components; it should not interfere with crucial protective immunological processes, but it may still alleviate parasitemia by positive immune modulation.

Leishmania infantum is the causative agent of human and canine visceral leishmaniasis (CVL) in the Mediterranean region. A seroprevalence study for CVL was conducted in northern Palestine. Domestic dogs (n = 148) were screened for antileishmanial antibodies by enzyme-linked immunosorbent assay (ELISA). Ten dogs (6.8%) were seropositive. Promastigotes were isolated from one seropositive dog and identified as L. infantum by excreted factor (EF) serotyping, isozyme electrophoresis, and polymerase chain reaction (PCR). In addition to the ELISA, the internal transcribed spacer 1 (ITS1)-, modified ITS1 (mITS1)-, and kinetoplast DNA (kDNA)-PCRs were used to validate this technique as a diagnostic tool for CVL using blood; each assay was performed on 60 blood samples. kDNA-PCR (13/60 positives, 21.7%) was the most sensitive of the assays examined followed by mITS1-PCR (9/60, 15.0%), ELISA (5/60, 8.3%), and ITS1-PCR (3/60, 5%). However, ITS1-PCR and mITS1-PCR were also capable of identifying the parasite species and indicated they belong to L. infantum. In view of its higher sensitivity, kDNA-PCR is recommended for the routine diagnosis of CVL.

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 225 free-range chickens (Gallus domesticus) from Portugal was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT) and found in 61 chickens with titers of 1:5 in 8, 1:10 in 6, 1:20 in 3, 1:40 in 23, 1:80 in 5, 1:160 in 4, 1:320 in 8, and 1:640 or higher in 4. Hearts, leg muscles, and brains of 15 seropositive (MAT 1:10 or higher) chickens were bioassayed individually in mice. Tissue from 38 chickens with titers of 1:5 or less were pooled and fed to a T. gondii–free cat. Feces of the cat were examined for oocysts, but none was found. Toxoplasma gondii was isolated from 16 of 19 chickens with MAT titers of 1:10 or higher. Genotyping of 12 of these 16 isolates with polymorphisms at the SAG2 locus indicated that 4 were type III, and 8 were type II. None of the isolates was lethal for mice. Phenotypically, T. gondii isolates from chickens from Portugal were different from those of T. gondii isolates from chickens from Brazil.

The helminth fauna of 76 Emydura macquarii from 3 river systems in central and northern Queensland was examined. Eleven species were found, including 2 nematodes, 6 trematodes, 1 aspidogastrean, 1 cestode, and 1 monogenean. Analysis of helminth diversity showed that the Fitzroy and Ross River turtles had communities of comparable diversity, but the helminth communities in Proserpine River turtles were much less diverse. The helminth communities in all localities were dominated by trematodes. Polystomoides australiensis was the most prevalent, being found in 60% of the Ross River turtles, 57% of the Fitzroy River turtles, and 46% of the Proserpine River turtles. Notopronocephalus peekayi was the most abundant species, with mean abundances of 5.9 in the Ross River turtles and 9.8 in the Fitzroy River turtles. Species richness, Simpson's Reciprocal Index, was highest, 4.68, for the Ross River helminth community, Sorensen's Qualitative Index showed 95% similarity between the Ross River and Fitzroy River communities, although Sorensen's Quantitative Index indicated only 35% similarity between the 2 sites. Host feeding patterns are likely the most important factor affecting species richness of the helminth infracommunities, as the majority of helminth species are transmitted by food-web interactions involving intermediate hosts.

Male preputial and female clitoral glands of mice undergo development that depends on the level of hormones in the animal. Experimental infection with Taenia crassiceps cysticerci results in significant physiological modifications in the host. Here, we investigated the histomorphological alterations induced by the parasite in these pheromonal glands. Preputial and clitoral glands were recovered from mice at 15, 35, 50, and 70 days postinfection (DPI). The glands were examined macroscopically and microscopically after histological preparation. Male preputial glands show a marked atrophy 35 days after infection. This atrophy is the result of a disorganization of the acinus tissue structure. During the course of infection, the basal, intermediate, and mature acinar cell layers are reduced, and finally, at 70 DPI, the gland includes only the duct system and fibrotic structures. In contrast, females are not affected by the infection because no modifications were observed in the morphology or histology of the clitoral glands. A probable cause for such a divergence between infected male and female mice might be related to a sex steroid imbalance as described during T. crassiceps infection.

Two pharyngodonid nematode species, Pharyngodon tiliquae and Thelandros trachysauri, infect the Australian lizard Egernia stokesii (gidgee skink) in populations from South Australia. Eggs are detected in lizard scats that are deposited in piles outside the rock crevice refuges that the lizards occupy. Eggs were isolated by salt flotation from fresh scats and from scats that had been dried in simulated field conditions for 7, 14, 21, and 28 days. Egg counts decreased with drying time for both nematode species, but T. trachysauri eggs were still detected after 28 days of drying, whereas P. tiliquae eggs were rarely detected after 14 days. These results suggest that egg counts can be used to infer host infection status only from relatively fresh scats and that eggs of the 2 species persist in a state where they can be detected by standard flotation techniques, for different times.

Ingestion of Toxoplasma gondii tissue cysts can result in severe disease in immunocompromised individuals and pregnant women. Treatment of meat and meat products to eliminate viable T. gondii tissue cysts would provide a means to protect consumers. In this study, we examined the effects of high-pressure processing (HPP) on ground pork containing viable tissue cysts of the VEG strain of T. gondii. Ground pork containing tissue cysts was exposed to 400, 300, 200, 100, or 0 MPa treatment for 30, 60, or 90 sec in a commercial HPP unit. The HPP-treated ground pork was subjected to acid–pepsin digestion and bioassayed in mice. The results of the mouse bioassay revealed that none of the mice inoculated with tissue cysts exposed to 400 or 300 MPa became infected, whereas all mice inoculated with tissue cysts exposed to 200, 100, or 0 MPa became infected with T. gondii regardless of exposure time. Results indicate that HPP treatment of ground pork with 300 MPa of pressure will render tissue cysts of T. gondii nonviable and make pork safe for human consumption.

We investigated whether variations in gene expression of enzymes associated with anaerobic resistance of laboratory-derived strains of Trichomonas vaginalis could be detected in a group of 28 clinical isolates with variations in metronidazole sensitivity. We compared isolates by real-time PCR because this method allows for highly sensitive quantification of mRNA and for evaluation of several genes simultaneously. We found that PFOR gene A mRNA levels were highly correlated with PFOR gene B levels, as well as the D subunit of malic enzyme and ferrodoxin. Ferrodoxin mRNA expression was also significantly correlated with that of malic enzyme and hydrogenase. However, when we evaluated relationships between these enzymes and resistance to metronidazole, we found no significant correlations between aerobic or anaerobic in vitro sensitivity to drug and mRNA levels of any of the enzymes tested. Similarly, using a Student's t-test, no significant differences in enzyme mRNA levels were observed between isolates separated by metronidazole resistance or susceptibility. The lack of correlation between gene expression and resistance or susceptibility could be the result of differences in expression at the protein level or because other biochemical pathways or genes are involved in the resistance observed in clinical settings.

The existence of latent Taenia solium post-oncospheral stages in the tissues of infected pigs has been postulated. To assess whether such structures exist and can be detected, we examined muscle samples from cysticercosis-infected and uninfected pigs. Pork samples were homogenized, centrifuged, and resuspended in saline solution. Round microscopic structures of approximately 10 μm with variable refringence were found in the pellets of all samples from both infected and uninfected pigs. These became homogeneously red after staining with Sudan IV and disappeared after ether extraction. The only difference between samples from infected and uninfected pigs was the presence of inflammatory cells and tissue necrosis debris in the former group. Taenia solium oncospheres were stained and observed for comparative purposes, before and after inoculation into pork. Control oncospheres were ellipsoidal, had nucleated basophile cells in their interior, and showed red aggregates on their surfaces when stained with 3% Sudan IV. While rounded microscopical structures similar to those previously reported were found, these differed morphologically from oncospheres, were of a lipid nature, and occurred in both infected and uninfected animals. No evidence supporting the presence of latent post-oncospheral stages of Taenia solium was generated in this series of experiments.

Sporozoites of 3 isolates of Plasmodium cynomolgi dissected from the salivary glands of Anopheles dirus and Anopheles quadrimaculatus were injected intravenously into 9 New World monkeys. Liver stage parasites were demonstrated in all 9 animals; 7 of these animals also produced blood stages after prepatent periods of 9 to 23 days.

We report on the results of radical surgery performed on a 10-yr-old Chinese female with multiple echinococcosis lesions and the diagnosis of the infection by imaging, histology, serology, and DNA analysis. Molecular genotyping provided unequivocal proof that the patient was infected with Echinococcus granulosus, the cause of cystic echinococcosis.

Actin is a ubiquitous and highly conserved microfilament protein that is hypothesized to play a mechanical force-generating role in the unusual gliding motility of sporozoan zoites and their active penetration of host cells. We have identified and isolated an actin gene from a Babesia gibsoni cDNA library by random sequencing. The complete nucleotide sequence of the actin gene is 1,243 bp; a single open reading frame encodes a polypeptide of 377 amino acid residues. The deduced amino acid sequence showed a high homology with actins from other species, especially with reported apicomplexan protozoans. The antiserum against recombinant actin expressed in Escherichia coli recognizes a 42-kDa native protein, which is consistent with its expected size. Immunofluorescence and confocal microscopic observation revealed that the protein is diffusely distributed throughout the B. gibsoni parasites.