Toxoplasma gondii oocysts are environmentally resistant and can survive outdoors for many months in dry and cold climates. In the present study, sporulation and survival of T. gondii oocysts was studied in different types of cat litters commercially available in the United States. Oocysts sporulated within 2–3 days in all types of cat litters and occasionally remained viable for 14 days. Results indicate that cat litter should be changed daily to prevent sporulation and infectivity to people.

Larval tapeworms (Polypocephalus sp.) reside within the central nervous system of decapod crustaceans. Living within the nervous system would seem to create an excellent opportunity for the parasites to manipulate the behavior of their hosts, so we tested the hypothesis that behavior of white shrimp (Litopenaeus setiferus) would be correlated with the level of parasitic infection. We videorecorded the behavior of L. setiferus for 8 hr, then examined the nervous system and digestive glands for parasite infection. Larval Polypocephalus sp. were found in the nerve cord, often in large numbers, but only very rarely in the digestive gland, which was typically infected by the larval stage of the nematode, Hysterothylacium sp. There were significantly more Polypocephalus larvae in the abdominal and thoracic ganglia than the subesophageal ganglia and brain. Walking, but not swimming, was significantly and positively related to the number of Polypocephalus sp. lodged in nervous tissue, as well as shrimp carapace length. Polypocephalus sp. is 1 of only a few parasites residing inside the host nervous system and it may, therefore, be suitable for investigating mechanisms of parasite manipulation of invertebrate host behavior.

This study was undertaken to identify the community composition, structure, and dynamics of helminths infecting the gastrointestinal tract (GIT) of lake whitefish (Coregonus clupeaformis) collected from 4 sites in northern lakes Huron (Cheboygan and De Tour Village) and Michigan (Big Bay de Noc and Naubinway) from fall 2003 through summer 2006. A total of 21,203 helminths was retrieved from the GITs of 1,284 lake whitefish. Approximately 42% (SE  =  1.4%) of the examined lake whitefish were infected with at least 1 helminth species in their GIT, with a mean intensity of 39.4 worms/fish (SE  =  0.3) and a mean abundance of 16.4 worms/fish (SE  =  0.1). Collected helminths appeared to be generalists and consisted of 2 phyla (Acanthocephala and Cestoda) and 5 species (Acanthocephalus dirus, Neoechinorhynchus tumidus, Echinorhynchus salmonis, Cyathocephalus truncatus, and Bothriocephalus sp.). Lake whitefish from Lake Huron on average had greater infection prevalences, abundances, and intensities than did fish from Lake Michigan. Infection parameters for each of the helminth species generally followed the same pattern observed for the combined data. Acanthocephalus dirus was the most prevalent and abundant helminth in lake whitefish GITs, although intensity of infection was the greatest for C. truncatus. Helminth infection parameters often peaked in the spring while diversity was greatest in the winter samples. There was substantial temporal variability in helminth infections with prevalences, abundances, and intensities often fluctuating widely on consecutive sampling occasions. Analysis of the GIT helminth community composition suggested that 3 (Big Bay de Noc, De Tour Village, and Cheboygan) of the 4 primary spawning sites, overall, had similar community compositions. The reason for the observed spatial and temporal variability in the lake whitefish GIT helminth infections remains to be elucidated. The findings of this study represent the most comprehensive parasitological study ever conducted on lake whitefish in the Great Lakes and will provide valuable information for future comparisons.

The diphyllobothriidean cestode Schistocephalus solidus typically infects threespine sticklebacks that are too small to allow the parasite to reach a mature size. As a result, the parasite must allow further growth of its host to reach the size at which it becomes competent to infect and reproduce in the definitive host. At times, however, intensity of infection can be high, leading to crowding among parasites and to heavy burdens causing mortality among hosts. Our data show that, during a previously observed epizootic, large percentages of plerocercoids (average 75% per host, 82% among all parasites pooled) did not grow to become massive enough in 1-yr-old threespine sticklebacks to be capable of establishment and maturation in the definitive host. Massive deaths of 1-yr-old sticklebacks due to infection during the epizootic resulted in the great misfortune of a disaster for a large number of parasites, resulting in dramatically reduced transmission of S. solidus.

The parasite fauna of the gray four-eyed opossum, Philander opossum (Linnaeus, 1758), and the common opossum, Didelphis marsupialis Linnaeus, 1758, in Camp du Tigre, French Guiana, is characterized. Nine species from the gastrointestinal system were recovered from both species, which shared 80% of their parasites. The parasite fauna comprised several monoxenous species (63%) and was dominated by Aspidodera raillieti Travassos, 1914, which exhibited high levels of prevalence and abundance in both communities. Only 2 species (Moennigia sp. and Spirura guianensis) had been recorded in other species of mammals. Both species richness and taxonomic composition at the level of component communities from this locality were compared against 11 communities present in the Virginia (Didelphis virginiana), white-bellied (Didelphis albiventris), and common opossum from Argentina, Brazil, Mexico, and the United States. Neither host phylogeny nor taxonomy accounted for statistical differences in species richness. There was no statistical difference among species richness values among the 9 localities studied. Taxonomic similarity was analyzed by means of the Jaccard's similarity index, including all, and only common species (occurring in prevalence >10%). The results suggest that sympatric species of marsupials share more species of parasites than parasite communities occurring in conspecific marsupials from different localities. As a consequence, taxonomic composition of these parasite communities varied depending on the locality. Probably, marsupials of the monophyletic Didelphini offer the same compatibility toward their parasites, by presenting them with similar habitats. Subtle differences in lifestyles of the marsupials may determine the chance of encounter between the symbionts and prevent some parasites from completing their life cycles. Further and more rigorous tests are necessary to determine the roles of encounter and compatibility filters, as well as the role of chance, in the structuring of parasite communities in marsupials.

Many biotic interactions can affect the prevalence and intensity of parasite infections in aquatic snails. Historically, these studies have centered on interactions between trematode parasites or between trematodes and other organisms. The present investigation focuses on the nematode parasite Daubaylia potomaca and its interactions with a commensal, Chaetogaster limnaei limnaei, and a variety of trematode species. It was found that the presence of C. l. limnaei indirectly increased the mean intensity of D. potomaca infections, apparently by acting as a restraint for various trematode parasites, particularly the rediae of Echinostoma sp. In turn, Echinostoma sp. rediae adversely affected the mean intensity of D. potomaca by their consumption of both juvenile and adult nematodes present in tissues of the snail. These organisms not only belong to 3 different phyla but occupy distinct trophic levels as well. The complex interactions among these 3 organisms in the snail host provide an excellent example of biotic interactions influencing the infection dynamics of parasites in aquatic snails.

Thirty-five deer mice, Peromyscus maniculatus, were trapped on Gannet Cluster 2 (GC-2), one of a group of islands numbered by convention in the Gannet Island Archipelago, and examined for ectoparasites. One species each of Acari (Ixodes uriae) and Siphonaptera (Orchopeas leucopus) were recovered. Samples of mice favored males to females (3.4∶1). Twenty-nine percent (10) of the mice were free of ectoparasites. Males were more heavily parasitized than females when both parasites were considered. No ticks were recovered from the female mice, while the males that were parasitized carried adult Ixodes uriae. These 2 ectoparasites parasitizing P. maniculatus, which is a known reservoir host for Lyme disease (Borrelia burgdorferi), may carry B. garinii and their presence would have serious implications for the spread of this human pathogen northward in continental North America.

Androlaelaps ulysespardinasi, a new species of laelapid mite, is described based on specimens collected from the pelage of the akodontine rodent Akodon philipmyersi Pardiñas, D'Elía, Cirignoli and Suarez, 2005, which is endemic to the Northern Campos Grasslands in Misiones Province, Argentina. The formal taxonomic description, illustrations, and scanning electron micrographs of the new species are derived from female specimens; males and nymphs are unknown. Androlaelaps ulysespardinasi resembles Androlaelaps rotundus Fonseca, 1936 in general appearance, but it differs in having a more sclerotized, V-shaped and elevated ridge among j4 and j6 setae in the dorsal shield; the idiosoma is smaller (586–634 µm in length) and has a bilobed posterior margin. The sternal plate is 1.3 times broader than longer, the epigynial shield is 1.2 longer than broader, and an anal shield is almost as long as it is broad. Both species are close to Androlaelaps misionalis Lareschi, 2010 and Androlaelaps maurii Lareschi and Gettinger, 2009, but they differ in their larger size (586–650 µm), in having the distance between j6 setae similar to the distance between j5 setae (72–83 µm), 11 pairs of setae in opisthogaster, and lacking a pair of setae close to epigynal shield. In addition, An. ulysespardinasi n. sp. is unusual in having chelicerae with both digits with 2 teeth. An An. rotundus species group, which includes the new species, is proposed, and a key to identify the different species is provided. Host specificity of the different species with rodents from the Akodon division of the akodontines was observed, suggesting that cophylogeny between these mites and akodontines may have occurred.

During surveys of helminth parasites of marine fishes from the Pacific coast of the Guerrero State in Mexico, 2 undescribed species of Haliotrema Johnston and Tiegs, 1922 (sensu Young, 1968) (Dactylogyridae) were found infecting the gills of the giant hawkfish Cirrhitus rivulatus Valenciennes, 1846 (Perciformes, Cirrhitidae), i.e., Haliotrema cirrhitusi n. sp. and Haliotrema pollexinus n. sp. The new species are similar with respect to the morphology of the anchors with base–shaft junction crazed, hinged, or both; dorsal anchors with an elongate superficial root; a tubular male copulatory organ (MCO) with a cylinder-shaped base diagonally opening; and a membranous accessory piece, shaped like a vane, encircling MCO to form 2 complete spirals from the anterior margin of base to the terminal portion of the MCO. While H. cirrhitusi and H. pollexinus differ from all other species of the genus in the general morphology of the copulatory complex, only H. pollexinus notably differs from all these species in having a superficial, blade-like projection from the inner surface on the distal shaft of anchors. Haliotrema cirrhitusi and H. pollexinus are the only members of the genus recorded in the Mexican Pacific and from a cirrhitid host.

The swimming behavior of the cercaria of the digenetic trematode Proterometra macrostoma changes in response to light. However, this cercaria does not possess obvious eyes or eyespots. Using behavioral assays, we were able to show that both intact and distome-removed cercariae swim significantly greater vertical distances under dim, red light than under brighter, white light. Electrophysiological experiments confirmed this result and further showed that the transverse band of the tail, known to control cercariae swimming behavior, was necessary and sufficient for the display of the light-dependent swimming behavior. Together, these data show that the distome is not required for light-dependent swimming behavior in P. macrostoma cercariae and indirectly demonstrates the presence of photoreceptors in the transverse band of the cercaria tail.

There is a lack of information concerning the prevalence of Toxoplasma gondii infection in wild birds in Mexico. In the present study, serum samples and tissues from 653 birds from Durango State, Mexico, were evaluated for T. gondii infection. Antibodies to T. gondii (modified agglutination test, titer 1∶25 or higher) were found in 17 (2.6%) of the 653 birds, including 1 of 2 curve-billed thrashers (Toxostoma curvirostre), 2 (1 Anas platyrhynchos, 1 Anas diazi) of 4 ducks, 1 of 2 eagles (Aquila sp.), 5 (27.8%) of 18 great-tailed grackles (Quiscalus mexicanus), 7 (1.3%) of 521 rock pigeons (Columba livia), and 1 (14.3%) of 7 quail (Coturnix coturnix). The seroprevalence of T. gondii infection in birds captured in a park outside the city zoo (11.6%, 8/69) was significantly higher than that found in birds from other regions (1.5%, 9/584, OR  =  8.38; 95% CI: 2.82–24.77; P  =  0.0001). Brains and hearts of 23 birds (17 seropositive, 6 seronegative) were bioassayed in mice for the isolation of T. gondii. Viable T. gondii was isolated from 1 of 7 seropositive pigeons. The DNA obtained from the T. gondii isolate from the pigeon was genotyped using the PCR-RFLP typing using 11 markers (B1, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and revealed an atypical genotype. This is the first report of T. gondii infection in great-tailed grackles, the Mexican duck, and curved-billed thrashers and the first survey of wild birds in Mexico.

Cholangiocytes, the epithelial cells lining intrahepatic bile ducts, express multiple toll-like receptors (TLRs) and, thus, have the capacity to recognize and respond to microbial pathogens. In previous work, we demonstrated that TLR4, which is activated by gram-negative lipopolysaccharide (LPS), is upregulated in cholangiocytes in response to infection with Cryptosporidium parvum in vitro and contributes to nuclear factor-kappaB (NF-kB) activation. Here, using an in vivo model of biliary cryptosporidiosis, we addressed the functional role of TLR4 in C. parvum infection dynamics and hepatobiliary pathophysiology. We observed that C57BL mice clear the infection by 3 wk post-infection (PI). In contrast, parasites were detected in bile and stool in TLR4-deficient mice at 4 wk PI. The liver enzymes alanine transaminase (ALT) and aspartate transaminase (AST), and the proinflammatory cytokines tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-6 peaked at 1 to 2 wk PI and normalized by 4 wk in infected C57BL mice. C57BL mice also demonstrated increased cholangiocyte proliferation (PCNA staining) at 1 wk PI that was resolved by 2 wk PI. In contrast, TLR4-deficient mice showed persistently elevated serum ALT and AST, elevated hepatic IL-6 levels, and histological evidence of hepatocyte necrosis, increased inflammatory cell infiltration, and cholangiocyte proliferation through 4 wk PI. These data suggest that a TLR4-mediated response is required for efficient eradication of biliary C. parvum infection in vivo, and lack of this pattern-recognition receptor contributes to an altered inflammatory response and an increase in hepatobiliary pathology.

Environmental stressors have the potential to greatly impact the transmission of parasites with complex, multi-host life cycles such as those of trematodes. The commonly used herbicide atrazine has been shown to affect the susceptibility of second intermediate hosts (such as larval amphibians) to trematode infection, as well as the longevity and infectivity of the free-swimming cercariae, but not eggs or the free-swimming miracidia that infect the gastropod first intermediate hosts. However, we do not know if this pesticide influences the survival of infected snails or whether it affects cercariae production within, or emergence from, these hosts. In addition, previous studies of host–parasite dynamics have only examined the parent atrazine compound, not any of the long-lasting metabolites commonly present in water bodies. Here, we report that a concentration of 0.33 µg/L of an atrazine metabolite, desethyl atrazine, increased the mortality of freshwater gastropods (Stagnicola elodes) infected with a gymnocephalus type of cercaria but not that of uninfected snails or those harboring a mature or dormant infection of Echinoparyphium sp. In contrast, 2 wk of exposure to desethyl atrazine did not affect the emergence of gymnocephalus cercariae from snails, although a trend for a decrease in the emergence of Echinoparyphium sp. cercariae was observed. We suggest that simultaneous trematode infection and exposure to contaminants may represent a significant combined stress to gastropods, but this is likely parasite species-specific as well as dependent on whether cercariae are being actively produced.

This work describes associations of Thiara (Tarebia) granifera, its larval trematode community, and Chaetogaster limnaei limnaei at a freshwater reservoir in Jamaica. Larvae of 2 trematodes were present, i.e., a notocotylid (15.3%) and Philophthalmus sp. (1.3%), in 3,575 T. granifera examined. The prevalence of both infections increased with snail shell length (H  =  56, P < 0.01, H  =  23.1, P < 0.01, respectively). Only 3.0% (n  =  595) of infected snails possessed reproductive stages, compared with 90.3% (n  =  2,980) of uninfected snails (χ²  =  2,059.8, df  =  1, P < 0.001); both trematodes negatively impacted snail reproduction. Chaetogaster l. limnaei occurred within the mantle cavity of T. granifera with a prevalence of 2.3% (n  =  3,575); intensity ranged from 1 to 6 annelids. Notocotylid larvae occurred in 32.5% (n  =  83) of snails also harboring C. l. limnaei, compared with 14.9% (n  =  3,492) of snails lacking the annelid (χ²  =  18.127; P < 0.001). Chaetogaster l. limnaei appears not to influence the recruitment of egg-transmitted, notocotylid infections to snails. Ingestion of emergent cercariae by the annelid was observed; this may impact transmission of the parasite. The article presents the first report of a notocotylid and C. l. limnaei in T. granifera, and of Philophthalmus sp. in Jamaica.

Adults of Lecithochirium floridense (Digenea: Hemiuridae) parasitized the stomach in each of 22 necropsied lionfish, Pterois cf. volitans (Scorpaeniformes: Scorpaenidae) (prevalence  =  100%, mean intensity  =  11), captured in the northwestern Atlantic Ocean off Beaufort, North Carolina (34°14.83′N, 76°35.25′W). This is the first report of a digenean from the invasive lionfish and that of L. floridense from a species of Pterois. The leech specimen previously identified as Myzobdella lugubris from P. volitans in the northwestern Atlantic Ocean is re-identified as Trachelobdella lubrica based on a study of the original voucher specimen.

Recently, Besnoitia neotomofelis was described from a southern plains woodrat (Neotoma micropus) from southern Texas. During May 2010, 1 of 55 southern plains woodrats trapped in Uvalde County, Texas, was diagnosed with besnoitiosis. Grossly, the woodrat had bilateral swellings of the cheeks, and numerous Besnoitia sp.-like cysts were observed in the tongue, facial region, musculature of the limbs, and subcutis of the dorsum and flanks. Little to no inflammation was noted around cysts. The cysts were morphologically similar to B. neotomofelis based on light and transmission electron microcopy. The sequence of the internal transcribed spacer region-1 was identical to the type isolate of B. neotomofelis. Attempts to isolate Besnoitia sp. in laboratory mice failed; however, Toxoplasma gondii was isolated in a Swiss Webster mouse. This represents the first report of besnoitiosis caused by B. neotomofelis in a southern plains woodrat and the first concurrent Besnoitia sp. and T. gondii infection in any host species.

Toxoplasma gondii was isolated from a feral guinea fowl (Numida meleagris) and domestic rabbits (Oryctologus cuniculus) from Brazil for the first time. Serum and brains from 10 guinea fowl and 21 rabbits from Brazil were examined for T. gondii infection. Antibodies to T. gondii were found in 2 of 10 fowl and 2 of 21 rabbits by the modified agglutination test (titer 1∶25 or higher). Viable T. gondii (designated TgNmBr1) was isolated from 1 of the 2 seropositive fowl by bioassay in mice but not from the 8 seronegative fowl by bioassay in cat. Viable T. gondii was isolated from both seropositive rabbits (designated TgRabbitBr1, TgRabbitBr2) by bioassay in mice from 1 and by bioassay in cat from the other. The TgRabbitBr1 strain was highly virulent for out-bred mice; mice fed 1 infective oocyst died of acute toxoplasmosis. The remaining 2 isolates were relatively avirulent for mice; lethal dose for mice was 10,000 oocysts. All 3 isolates were grown in cell culture, and tachyzoite-derived DNA were genotyped using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The TgNmBr1 was found to be clonal Type II, a rare finding in Brazil in any host. The rabbit isolates were atypical, similar to isolates from cats from Brazil (TgRabbitBr1 was identical to TgCatBr5, and TgRabbitBr2 was identical to TgCatBr1, a common genotype in Brazil denoted type BrII). This is the first genetic characterization of T. gondii isolates from the rabbits and guinea fowl in Brazil and the first host record for T. gondii in the guinea fowl.

Because the taxonomy of trematodes is based on adults, the larval stages of most digeneans cannot be identified to species based on morphology alone. Molecular data provide a means of linking larval stages to known adults. We obtained sequences from the barcode region of cytochrome oxidase I (CO1) from adult and larval parasites of fish, frogs, birds, and mammals across North America. Sequences from adult Apharyngostrigea cornu, Hysteromorpha triloba, and Alaria mustelae (Diplostomoidea: Digenea) from definitive hosts matched those of meta- and mesocercariae from fish and frogs. These data provided new information on the distributions of all 3 parasite species. Metacercariae of A. cornu, which have not been previously reported in North American hosts, were found in Notemigonus crysoleucas, Pimephales notatus, and Catostomus commersonii in the St. Lawrence River. Metacercariae of H. triloba are reported in Canadian waters and in N. hudsonius for the first time. Alaria mustelae is reported for the first time in frogs from Quebec, Canada, and an additional species of Alaria was detected in California. Sequences of internal transcribed spacer rDNA from a subset of specimens support the same species boundaries indicated by CO1 divergence. There was little divergence in CO1 sequences from an unidentified diplostomid species sampled at a large spatial scale.

It is not unusual to find common molecules among parasites of different species, genera, or phyla. When those molecules are antigenic, they may be used for developing drugs or vaccines that simultaneously target different species or genera of parasite. In the present study, we used a proteomic-based approach to identify proteins that are common to adult Fasciola hepatica and Schistosoma mansoni. Whole-worm extracts from each parasite were separated by 2-dimensional electrophoresis (2-DE), and digital images of both proteomes were superimposed using imaging software to identify proteins with identical isoelectric points and molecular weights. Protein identities were determined by mass spectrometry. Imaging and immunoblot analyses identified 28 immunoreactive proteins that are common to both parasites. Among these molecules are antioxidant proteins (thioredoxin and glutathione-S-transferase), glycolytic enzymes (glyceraldehyde 6-phosphate dehydrogenase and enolase), proteolytic enzymes (cathepsin-L and -D), inhibitors (Kunitz-type, Stefin-1), proteins with chaperone activity (heat shock protein 70 and fatty acid–binding protein), and structural proteins (calcium-binding protein, actin, and myosin). Some of the identified proteins could be used to develop drugs and vaccines against fascioliasis and schistosomiasis.

Previously, we reported a tick recovered from Antelope Cave in extreme northwest Arizona. Further analyses of coprolites from Antelope Cave revealed additional parasitological data from coprolites of both human and canid origin. A second tick was found. This site is the only archaeological locality where ticks have been recovered. We also discovered an acanthocephalan in association with Enterobius vermicularis eggs in the same coprolite. This association shows that the coprolite was deposited by a human. This discovery expands our knowledge of the range of prehistoric acanthocephalan infection. In addition, findings from canid coprolites of Trichuris vulpis are reported. This is the first published discovery of T. vulpis from a North American archaeological context. The close association of dogs with humans at Ancestral Puebloan (Anasazi) sites raises the potential that zoonotic parasites were transferred to the human population. The archaeological occupation is associated with the Ancestral Pueblo culture 1,100 yr ago.

Enteric protozoal infection was identified in 5 stranded California sea lions (Zalophus californianus). Microscopically, the apical cytoplasm of distal jejunal enterocytes contained multiple stages of coccidian parasites, including schizonts with merozoites and spherical gametocytes, which were morphologically similar to coccidians. By histopathology, organisms appeared to be confined to the intestine and accompanied by only mild enteritis. Using electron microscopy, both sexual (microgametocytes, macrogamonts) and asexual (schizonts, merozoites) coccidian stages were identified in enterocytes within parasitophorous vacuoles, consistent with apicomplexan development in a definitive host. Serology was negative for tissue cyst-forming coccidians, and immunohistochemistry for Toxoplasma gondii was inconclusive and negative for Neospora caninum and Sarcocystis neurona. Analysis of ITS-1 gene sequences amplified from frozen or formalin-fixed paraffin-embedded intestinal sections identified DNA sequences with closest homology to Neospora sp. (80%); these novel sequences were referred to as belonging to coccidian parasites “A,” “B,” and “C.” Subsequent molecular analyses completed on a neonatal harbor seal (Phoca vitulina) with protozoal lymphadenitis, hepatitis, myocarditis, and encephalitis showed that it was infected with a coccidian parasite bearing the “C” sequence type. Our results indicate that sea lions likely serve as definitive hosts for 3 newly described coccidian parasites, at least 1 of which is pathogenic in a marine mammal intermediate host species.

The life cycle of Hepatozoon hemprichi n. sp. is described; the vertebrate host is Scincus hemprichii and it is vectored by Hyalomma impeltatum. Erythrocytic stages of 18 ± 1.8 × 4 ± 0.8 µm developed in the hemocoel of ticks to sporozoites within 16–18 days. Schizogony occurred in the liver parenchyma and the endothelial cells of blood capillaries in lung and spleen. Mature schizonts measuring 27 ± 3.11 × 20.13 ± 3.0 µm produced 28 merozoites (on average). The merozoites were 13 ± 1.21 × 1.21 ± 0.72 µm with nuclei 5 ± 0.65 × 2.1 ± 0.51 µm. Syzygy and differentiation of gamonts took place in tick's hemocoel up to the third day post-infection (PI). The microgamont (16 ± 0.31 × 18 ± 0.42 µm) produced 4, uniflagellated microgametes at 4–5 days PI. The microgamete measured 15.2 ± 0.31 µm while the flagellum was always at least 26 µm. The macrogamete was very large in size (31 ± 3.11 µm) with a central nucleus. After fertilization, (5–6 days PI) zygotes developed into oocysts (55 ± 3.41 × 52 ± 4.11 µm) in which repeated mitotic divisions with centripetal invaginations occurred; each contained 18 banana-shaped sporozoites, 13.61 ± 0.8 × 1.2 ± 0.31 µm in size. Experimental transmission was successfully carried out by oral administration or by intra-peritoneal inoculation of the infective stages (sporozoites) to uninfected skinks and led to the appearance of blood stages after 5 wk and 4 wk, respectively.

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum, by ∼1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.

Coccidiosis is an important disease in captive gamebirds, including northern bobwhites (Colinusvirginianus). Three Eimeria species, Eimeria lettyae, Eimeria dispersa, and Eimeria colini, have been described in bobwhites. Distinguishing the various Eimeria spp. is often problematic because of similarity in oocyst morphology and site of infection and thus requires live bird infections to distinguish between the coccidian species. To aid in identification and diagnosis, PCR specific primers were generated against the internal transcribed spacer region 1 (ITS-1) of the ribosomal RNA gene using sequences obtained from coccidian-positive samples collected from diagnostic cases or litter from captive bobwhites. Three distinct Eimeria spp. were detected. Species-specific primers were constructed and used to survey the prevalence of the species in 31 samples collected from 13 states. The primers survey results identified E. lettyae, E. dispersa, and Eimeria sp. in 20 (64.5%), 22 (71%), and 29 (93.5%) of the samples, respectively. Mixed infections were common: 13 (41.9%) samples had 3 Eimeria spp., 14 (45.2%) had 2 spp., and 4 (12.9%) samples had only 1 species. The species were widely distributed over the area sampled and were not associated with the age of the flock.

During November 2009 and March 2010, 20 adult eastern pipistrelles, Perimyotis ( = Pipistrellus) subflavus, were collected from Polk County, Arkansas, and their feces were examined for coccidian parasites. Two (10%) of the bats were found to be passing oocysts of an undescribed species of Eimeria. Oocysts of Eimeria heidti n. sp. were ovoidal to ellipsoidal, 26.1 × 20.5 µm (23–31 × 18–23 µm), with a bilayered wall, externally rough, internally smooth, and with a shape index of 1.3. Micropyle and oocyst residuum were absent, but a subspherical polar granule was often present. Sporocysts were ovoidal, 13.0 × 8.8 µm (11–15 × 7–13 µm), the shape index was 1.6, a Stieda body was present and sub-Stieda and para-Stieda bodies were absent. A sporocyst residuum consisting of multiple globules dispersed along the perimeter of the sporocyst and between the sporozoites were present, sporozoites were elongate, with a subspherical anterior refractile body and elongate posterior refractile body; a nucleus not discernible. This is the second coccidian reported from this host and the fourth instance of a coccidian species reported from an Arkansas bat.

The stinging catfish Heteropneustes fossilis (Bloch) (Siluriformes: Heteropneustidae) has been reported to harbor as many as 19 species of caryophyllidean tapeworms (Cestoda) of 11 genera in tropical Asia (Indomalayan zoogeographical region). However, a critical review of the species composition has shown that only 1 species, Lucknowia fossilisi Gupta, 1961 (Lytocestidae), is a specific parasite of H. fossilis. Three other species, Djombangia penetrans Bovien, 1926 (syn., Djombangia caballeroi Sahay and Sahay, 1977), Pseudocaryophyllaeus ritai Gupta and Singh, 1983 (syn. Pseudocaryophyllaeus lucknowensis Gupta and Sinha, 1984), and Pseudocaryophyllaeus tenuicollis (Bovien, 1926) Ash, Scholz, Oros and Kar, 2011 (syn. P. mackiewiczi Gupta and Parmar, 1982), were found only once. Lucknowia fossilisi is redescribed on the basis of new material collected in West Bengal and voucher specimens from Maharashtra, India. A total of 9 species of Capingentoides, Lucknowia, Lytocestus, Pseudoadenoscolex, Pseudocaryophyllaeus, Pseudoheteroinverta, and Sukhapatae are newly synonymized with L. fossilisi and previous synonymies of 9 other species, proposed by Hafeezulah (1993), are confirmed. Generic diagnosis of Lucknowia Gupta, 1961 is amended. In addition, 1 species of Pseudobatrachus and 2 species of the monotypic genera Pseudoneckinverta and Sudhaena are invalidated as nomina nuda.

Anisakis spp. larvae are parasitic, and potentially zoonotic, nematodes transmitted by marine fish and cephalopods, which are the main intermediate hosts of the third larval stage. The accidental consumption of infected raw or poorly cooked fish may cause gastroenteric diseases and allergies in humans. The aim of the present study was to use polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) to define the occurrence, species variability, and host preferences of Anisakis spp. larvae in fish caught off the coast of Sardinia. Necropsy was used on 285 samples; 552 Anisakis spp. L3 larvae were isolated from 87 fish that tested positive for this nematode. Anisakis pegreffii was most frequently encountered (90.6%), with a primary preference for Scomber scombrus, Zeus faber, and Trachurus mediterraneus. In contrast, the prevalence of Anisakis physeteris was only 1.3%. A hybrid genotype of Anisakis simplex sensu stricto and Anisakis pegreffii was also observed, which confirms the results of previous studies carried out in the western Mediterranean. Interestingly, no Anisakis simplex s.s. larvae were recovered. These results indicate that the diversity of Anisakis species is low in Sardinia waters, probably because of its geographic position.

Tetrathyridia of Mesocestoides sp. were collected from the body cavities of the shrews (Insectivora), Crocidura russula, in Valencia, Spain and Crocidura suaveolens on the Mediterranean island of Corsica, France. Specimens were processed by routine microscopic and histological techniques, including examination with brightfield, phase-contrast, and differential-interference-contrast optics. Most tetrathyridia were clustered together inside host-derived fibrotic capsules, but some occurred free in the body cavity. All specimens examined from both locations had solid hindbodies, i.e., lacking a primary lacuna, thus conforming to the plerocercoid metacestode type; all possessed a single normal tetra-acetabulate scolex. All metacestodes from C. russula in Valencia were normal tetrathyridia. Those from C. suaveolens in Corsica were either normal tetrathyridia or had aberrant deep convolutions of an unusually elongated hindbody. No tetrathyridium from either location or host showed tegumental or excretory duct anomalies such as those reported by several authors from aberrant tetrathyridia and spargana in some other locations. No definitive evidence of asexual proliferation was visible in any of the tetrathyridia, but those with abnormally convoluted hindbodies from a single C. suaveolens in Corsica suggest the potential for asexuality by fission of the hindbody. These results add to our understanding of morphological and developmental variation among metacestodes in this widespread and variable genus.

Macroderoides minutus n. sp. is described based on specimens collected from the Florida gar (Lepisosteus platyrhincus) from Orange Lake, Alachua County, Florida. The new species is the ninth recognized species of Macroderoides and is morphologically closest to Macroderoides flavus and Macroderoides typicus. It differs from both species in having a smaller body, ventral sucker larger than the oral sucker, substantially lower body length to width ratio, fewer and larger eggs, and other characters. A comparison of sequences of nuclear rDNA (partial 18S, complete ITS region, and partial 28S) of the new species, with sequences of 5 other Macroderoides, convincingly supports the status of M. minutus as a new species. Molecular phylogenetic analysis, including all North American Macroderoides species except for Macroderoides parvus, has demonstrated the presence of 2 distinct clades, 1 including Macroderoides spinifer (Macroderoides minutus n. sp. Macroderoides texanus) and the other including Macroderoides trilobatus (M. typicus M. flavus). The tree topology suggests that the similarity in body proportions among M. minutus n. sp., M. typicus M. flavus is not a result of close phylogenetic relationship. It has also revealed for the first time the phylogenetic affinities of M. flavus, which is the only North American member of Macroderoides that switched from holostean fishes to teleosts.

Trypanosoma anguiformis n. sp. and Trypanosoma polygranularis n. sp. are described from the African olive sunbird, Cyanomitra olivacea, and Latham's forest francolin, Francolinus lathami, respectively, based on the morphology of their hematozoic trypomastigotes and partial sequences of the small subunit ribosomal RNA gene. Both new species belong to the group of small non-striated avian trypanosomes (<30 µm in length on average) with the kinetoplast situated close to the posterior end of the body. Trypanosoma anguiformis can be readily distinguished from other small avian trypanosomes due to its markedly attenuated (snake-shaped) form of the hematozoic trypomastigotes and the dumbbell-shaped nucleus of the parasite. Trypanosoma polygranularis is readily distinguishable due to the markedly off-center (anteriorly) located nucleus, numerous azurophilic granules that are arranged in a line following the undulating membrane, and the large kinetoplast (with an area up to 1.7 µm² [1.1 µm² on average]). Illustrations of hematozoic trypomastigotes of the new species are given, and DNA lineages associated with these parasites are reported. The current situation in species taxonomy of avian trypanosomes is discussed. We call for the redescription of valid species of avian trypanosomes from their type vertebrate hosts and type localities by using morphological and polymerase chain reaction-based techniques as an initial essential step towards revising the species composition of avian trypanosomes and reconstructing the taxonomy of these organisms.

Based on light and scanning electron microscopical examinations, 2 new morphologically similar species of the subgenus Procamallanus (Spirocamallanus) (Nematoda: Camallanidae) possessing 2 caudal spikes are described from the intestine of marine perciform fishes from the Gulf of Thailand off the Thai coast, i.e., Procamallanus (S.) rigbyi n. sp. from Otolithes ruber (Bloch and Schneider) (Sciaenidae) and Procamallanus (S.) similis n. sp. from Sillago sihama (Forsskål) (Sillaginidae). The former species is mainly characterized by the presence of 13–14 spiral ridges in the buccal capsule, location of deirids at about mid-way between the buccal capsule and the nerve ring, and by the length of the right spicule (315–360 µm), whereas the latter is characterized by 10–12 spiral ridges, deirids situated at short distance posterior to the buccal capsule, and the right spicule 435–492 µm long. Because of their inadequate descriptions, most species of this subgenus reported from marine fishes of the Indo-Pacific region are considered species inquirendae. Spirocotyle Yasmin and Bilqees, 2007 is considered a junior synonym of Procamallanus Baylis, 1923. Since Procamallanus (S.) otolithi Ashraf, Farooq and Khanum, 1977, P. (S.) otolithi (Gupta and Garg, 1986) and P. (S.) otolithi (Yasmin and Bilqees, 2007) are junior homonyms to P. (S.) otolithi Bilqees and Kazmi, 1974, they are renamed Procamallanus (S.) pakistanensis n. nom., Procamallanus (S.) problematicus n. nom., and Procamallanus (S.) incognitus n. nom., respectively (all species inquirendae). Procamallanus rigbyi and P. similis are the first nominal species of this genus reported from marine fishes off the coast of Thailand.

The pfmdr1 gene, which encodes P-glycoprotein homolog 1, has been shown to be a reliable marker of resistance for Plasmodium falciparum related to artesunate and mefloquine combination therapy. The aims of this study are to investigate the copy number of pfmdr1 in P. falciparum isolates collected from the 4 malaria-endemic areas of Thailand (Kanchanaburi, Mae Hongson, Ranong, and Tak) along the Thailand–Myanmar (Burma) border (Thai–Myanmar border) by using SYBR Green I and the standard method TaqMan real-time polymerase chain reaction (RT-PCR) and to compare the efficiency (sensitivity and specificity) of SYBR Green I with TaqMan RT-quantitative (q)PCR methods in determining pfmdr1 gene copy number. Ninety-six blood samples were collected onto filter paper from patients with uncomplicated falciparum malaria who attended malaria clinics in the Kanchanaburi (n  =  45), Mae Hongson (n  =  18), Ranong (n  =  11), and Tak (n  =  22) provinces in Thailand. Parasite genomic DNA was extracted from dried blood spots by using QIAcube™ automated sample preparation. Pfmdr1 gene copy number was determined by TaqMan (63 samples) and SYBR Green I (96 samples) real-time PCR. Seventy-one (74.0%), 14 (14.6%), 10 (10.4%), and 1 (1%) isolates carried 1, 2, 3, and 4 pfmdr1 gene copies, respectively. Forty-three of 48 (89.6%), 6 of 11 (54.5%), and 3 of 4 (75.0%) samples, respectively, showed agreement with results of 1, 2, and 3 pfmdr1 gene copies as determined by both methods. The efficiency of SYBR Green I in identifying pfmdr1 gene copy number was found to be significantly correlated with that of TaqMan. Considering its simplicity and relatively low cost, SYBR Green I RT-qPCR is therefore a promising alternative technique for the determination of pfmdr1 copy number.

The aim of the present study was to determine the seroprevalence of Toxoplasma gondii in horses used for sporting purposes in the Province of Riyadh, Saudi Arabia. In total, 266 serum samples from clinically healthy horses were analyzed for anti–T. gondii antibodies using the Sabin-Feldman dye test. Antibodies to T. gondii were found in 84 (31.6%) horses, with specific titers of 1∶16 (78 with a prevalence of 29.3%), 1∶64 (4 with a prevalence of 1.5%), and 1∶256 (2 with a prevalence of 0.8%). The number of seropositive horses in Shaqra (43.7%) was considerably higher than in other regions, with the lowest occurring in Al-Majmaah (11.0%). However, the differences in seroprevalence between the 6 locations were not statistically significant (P > 0.05). In contrast, the overall seroprevalence in Riyadh province was higher than that reported in other countries. Horses are not a direct source of infection for human toxoplasmosis in this region. Usually human infection is facilitated via cats, which are the reservoir hosts for this parasite.

Here we provide the first North American report of a naturally infected snail, Gyraulus parvus, harboring the larval stages of the cosmopolitan, arterial schistosome, Dendritobilharzia pulverulenta. The relatively small cercariae of this species are shed in the early morning, are sticky, and adhere to the water's surface film. We also provide a report of the snail host, Physa gyrina, of the widespread North American passerine schistosome, Gigantobilharzia huronensis. Finally, we provide unambiguous documentation that Physa gyrina is a natural snail host for Trichobilharzia querquedulae, a schistosome primarily of dabbling ducks.

Prevalence of Toxoplasma gondii infection in pregnant women in Grenada is considered high. Little is known of the epidemiology of T. gondii infection in Caribbean Islands. Serum samples of 750 food animals in Grenada and Carriacou were tested for antibodies to T. gondii by the modified agglutination test (MAT). Antibodies to T. gondii (MAT, 1∶25 or higher) were found in 23.1% of 247 pigs, 44.1% of 204 sheep, 42.8% of 180 goats, and 8.4% of 119 cattle. Seroprevalence increased with age, indicating postnatal acquisition of T. gondii. Antibody titers of 1∶200 or higher were present in 65 of 90 seropositive sheep, 61 of 77 seropositive goats, and 23 of 57 seropositive pigs. However, none of the cattle had a MAT titer of 1∶200, suggesting that bovines are a poor host for T. gondii. Results indicate that pigs, sheep, and goats could be important sources of T. gondii infection if their meat is consumed undercooked.

Feces were collected from 68 dairy cattle, 1 to 12 mo of age, on 12 farms in the municipality of Campos dos Goytacazes, Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium sp. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) of the 18S rRNA. Four positive samples (4.54%) were sequenced and identified as Cryptosporidium andersoni. This species represents a risk for Brazilian cattle because infection can affect cattle productivity. Moreover, C. andersoni is considered a zoonotic species.

The genus Nonnapsylla Wagner, 1938 (Craneopsyllinae) includes only 1 species, Nonnapsylla rothschildi Wagner, 1938. Two subspecies are recognized by the shape of the dorsal margin of the helmet, apical shape of the spines in helmet and genal combs, length of first and fourth segments of the maxillary palpus, and shape of the hilla of the spermatheca. The validity of the characters used to separate the 2 subspecies of Nonnapsylla rothschildi Wagner, 1938 is discussed based on information from the literature, as well as from direct observation of type specimens and specimens collected in northwestern Argentina. We conclude that Nonnapsylla rothschildi wagneri Johnson, 1957 should be considered as a synonym of the nominal subspecies. We also extend the southern limits of the distribution of the species and report its presence in Argentina.

Antibodies to Toxoplasma gondii were found in 92 (9.9%) of 929 feral raccoons (Procyon lotor) in Japan with the use of the latex agglutination test (LAT, 1∶64 or higher). Seropositivity varied by geographic location, season, and weight of raccoons trapped. Seroprevalences in the northern, central, and western areas of Japan were found to be 7.9% (39/492), 16.5% (47/285), and 3.9% (6/152), respectively. The seroprevalence by season varied from 8.5% (13/153) in spring to 18.9% (14/74) in winter, which was significantly higher than those in other seasons (P < 0.05). Seroprevalence of T. gondii was elevated in accordance with the increase in body weight of raccoons (r_s  =  0.9), suggesting that the animals acquired the infection postnatally. The results suggest that the raccoon may serve as a useful indicator for the distribution of T. gondii in peridomestic environments in Japan.

The parasitic nematode Myrmeconema neotropicum infects workers of the neotropical arboreal ant Cephalotes atratus. Infected ants exhibit altered behavior, e.g., reduced aggression and slower tempo, as well as physical traits, e.g., gaster changes from shiny black to bright red. These changes are thought to induce fruit mimicry and attract frugivorous birds, which are the presumed paratenic hosts for the nematodes. We used respirometry to measure the energetic costs of nematode infection, testing the prediction of higher metabolic rates for infected workers maintaining both ant and nematode biomass. Contrary to this prediction, infected workers had lower mass-specific metabolic rates than uninfected workers. Parasites are limited to the gasters (abdomens) of adult ants, and infected gasters had 57% more mass, but 37% lower metabolic rates, compared to uninfected gasters. These results use a metabolic currency to measure, in vivo, the energetic costs of parasitism, and they shed light on the complex co-evolutionary relationship between host and parasite.

DNA samples from 737 tick pools, representing 6,850 Haemaphysalis longicornis and 51 Dermacentor nuttalli collected from Hebei Province, China, were analyzed by polymerase chain reaction (PCR) for the presence of spotted fever group Rickettsia. Fifty (6.9%) of 724 H. longicornis in the tick pool were positive, but no positive samples were found in 13 D. nuttalli. Sequence analysis of the partial outer membrane protein A (ompA) genes from the 10 positive samples showed 97.4–99.8% identity, but were different from the homologous sequence of Rickettsia previously deposited in GenBank. Phylogenetic analysis of ompA genes indicated that the Rickettsia detected in this study belonged to a novel haplotype, and formed a clade distinct from Rickettsia heilongjiangii, Rickettsia sibirica, and Rickettsia hulinii in China. The new strain, named Candidatus Rickettsia hebeiii, appears to represent a distinct lineage and could constitute a new species with a minimum prevalence of about 0.7% in H. longicornis from Hebei Province, China.