Circadian egg production by Echinostoma caproni was investigated in ICR mice. Four female mice were infected with 25 E. caproni metacercariae, maintained in individual cages on a 12:12 light:dark cycle, and provided food and water ad libitum. Twenty-eight, 51, and 58 days post-infection, mice were transferred to individual, wire-bottomed cages and feces were collected every 2 hr for 24 hr. The feces were weighed and processed immediately to estimate the number of eggs present. Fecal output and egg production were standardized to unit maxima for analysis. Standardized egg count and standardized fecal output followed distinctly circadian patterns and covaried. Egg production was highest from 2200 to 0200 hr and lowest from 1000 to 1800 hr. These correspond to the highest and lowest fecal production, and highest and lowest periods of host activity, respectively. Egg density (eggs/g of feces) covaried weakly with fecal output with an additional peak at 0800–1000 hr, suggesting E. caproni is responding to changes in host physiology in timing of the production and release of eggs into the intestine. The continuous production and release of eggs during the patent period, coupled with the circadian pattern of daily egg release by E. caproni, would result in the widest dispersal of eggs in the host environment and enhance transmission to the first intermediate host.

Gyrodactylus leptorhynchi n. sp. (Monogenea) is described from bay pipefish (Syngnathus leptorhynchus) (Syngnathidae) in coastal waters of southern California and British Columbia, and from an outbreak of gyrodactylosis at the Cabrillo Marine Aquarium in California. Gyrodactylus leptorhynchi is morphologically similar (stout hamuli, superficial bar with no anterolateral processes, and a small, triangular membrane, similarly shaped marginal hook sickles, and a male copulatory organ [MCO] with numerous small spines) to the other 6 species of Gyrodactylus known from pipefish in north and south regions of the Atlantic Ocean. It resembles most closely Gyrodactylus corleonis Paladini, Cable, Fioravanti, Faria, and Shinn, 2010, parasitizing Syngnathus typhle L. from the French Mediterranean in having relatively large hamuli (58 μm). However, in G. leptorhynchi, the marginal hook sickle has a reduced heel and a ledged toe, while in G. corleonis, it has a noticeable heel and a toe with no distinct ledge. DNA sequence data of a partial ITS1 (700 bp), complete 5.8S (157 bp), complete ITS2 (392 bp), and a partial 18S (441 bp) are included in the description; the data are distinct from those available for other species of Gyrodactylus. The molecular data reveal that G. leptorhynchi is a member of a basal lineage of marine species within Gyrodactylus sensu lato that is known to have radiated among coastal syngnathid, anguillid, and gobiid fishes throughout the Atlantic Ocean and some adjacent waters. Occurrence of G. leptorhynchi in the eastern Pacific supports the idea that such lineages may have global distributions. Sixty-three percent (15 of 24) of bay pipefish caught at Inner Cabrillo Beach, California, were infected with 1–3 worms, predominately located on the dorsal fin, but also on the anterior body surfaces. Intensely infected pipefish at the marine aquarium had parasites distributed all over the body surface, including the open edge of the brood pouch and, on 2 occasions, inside the brood pouch. A quarantine protocol, involving the treatment of wild pipefish with serial repeats of topical anthelmintic chemicals (formalin, Trichlorfon, and Praziquantel), that helps to diminish outbreaks of G. leptorhynchi in aquarium exhibits is described.

Laelapine mites are common parasites of sigmodontine rodents in the Neotropics. However, few species are reported from Peru as a result of the low number of mammal surveys that include ectoparasite collections. Herein we report 12 species of mites from northern Peru. From these, 8 are reported for the first time for the country, and 1 is new to science, Androlaepaps aerosus sp. nov., the latter associated exclusively with the sigmodontine Akodon aerosus. Most of the laelapine species were host specific. The new species, included in the Androlaelaps rotundus species group, resembles An. rotundus “sensu stricto” and An. ulysespardinasi in general appearance but is unique in the length of the hypostomal seta h3 (>58 μm), which is 3 times as long as the gnathosomal seta, and its tip reaching or over-reaching the gnathosomal setal bases; dorsal seta j2 is very long (>70 μm), almost reaching the point of j3.

Schistosoma mansoni glyceraldehyde 3-phosphate dehydrogenase (SG3PDH), peroxiredoxin (TPX), and other larval excretory–secretory products (ESP) essentially induce T helper (Th) 1 and Th17 immune responses during a non-protective natural infection. Such an immune environment promotes production of nitric oxide and hydrogen peroxide by interferon-γ–activated monocytes and interleukin (IL)-17–mediated recruitment and activation of neutrophils; however, it also likely prevents engagement of eosinophils and basophils in the hunt for developing larvae. We reasoned that polarizing ESP-induced immune responses toward a Th2 phenotype, via the use of cysteine proteases or type-2 cytokines, would lead to almost total parasite elimination. Accordingly, outbred mice were immunized with 10 μg recombinant SG3PDH and 15 μg TPX-derived peptide together with 10 μg papain, or 200 ng thymic stromal lymphopoietin, IL-25, or IL-33 as an adjuvant. Two weeks later, untreated mice, adjuvant controls, and immunized mice were challenged with 100 or 125 cercariae. Results of 6 experiments indicated that these formulations elicited IgM, IgG1, and IgA specific antibodies, and an increase in ex vivo spleen cells release of IL-4 and IL-5 correlated with highly significant (up to P < 0.0001) reduction of 62 to 78% in challenge worm burden. Improvement of ESP selection, singly or in a combination, and immunization regimen, namely ESP and type-2 cytokine dose and injection site and schedule, could lead to a sterilizing schistosomiasis vaccine in the foreseeable future.

Two new species of dicyemid parasites from Dicyema are described from 2 species of Australian cephalopods, i.e., Dicyema calamaroceum n. sp. from Sepioteuthis australis Quoy and Gaimard, 1832 (southern calamary) collected from Spencer Gulf (SG) and Gulf St Vincent (GSV), South Australia (SA), Australia, and Dicyema pyjamaceum n. sp. from Sepioloidea lineolata Quoy and Gaimard, 1832 (striped pyjama squid), collected from SG, SA, Australia. Dicyema calamaroceum is a medium sized species that reaches approximately 2,400 μm in length. The vermiform stages are characterized by having 31–34 peripheral cells, a conical calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos, and verruciform cells were not observed in nematogens and rhombogens. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell, and the refringent bodies are solid. Dicyema pyjamaceum is smaller than D. calamaroceum, with a body length that reaches approximately 1,950 μm. The vermiform stages are characterized by having 20–23 peripheral cells, a cap-shaped calotte that forms a cephalic swelling together with the parapolar cells, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Verruciform cells and granules in propolar cells were observed in nematogens and rhombogens. Infusoriform embryos consist of 37 cells; 2 nuclei are present in each urn cell, and the refringent bodies are solid. This represents the first description of dicyemid parasites from Australia.

Water mites of Unionicola species are common symbionts of freshwater mussels, living on the gills or mantle and foot of their hosts and using these tissues as sites of oviposition. Although surveys of the mite fauna among North American mussels suggest that these mites represent highly diverse assemblages, there are currently no quantitative data characterizing Unionicola species diversity among their molluscan hosts. The present study addresses patterns of species richness of Unionicola assemblages from freshwater mussels, including the relationship between richness and host specificity among these mites. Results from this study indicate that mite species richness increased significantly with an increase in the number of host individuals sampled. When corrected for sampling effort, there was a positive relationship between host size and mite species richness. Results from this study also reveal a significant relationship between mite species richness and the geographical distribution of host mussels. Overall, the patterns of species richness observed for this study are consistent with those examining the richness of parasitic helminth communities. Because the phylogenetic history of host taxa can have a significant influence on patterns of parasite species richness, studies that correct for the phylogenetic history among host mussels will be required to better understand the role that evolutionary processes have in determining Unionicola species richness. The present study did not indicate a significant relationship between species richness and host specificity and, in not doing so, suggests that the dispersal ability of mites may also play a role in influencing Unionicola species richness. The host recognition behavior and swimming abilities for a larger sample of mites will be required to substantiate this hypothesis.

We elucidate the life cycle of Maritrema orensense for the first time and experimentally confirm that of the sympatric Maritrema bonaerense. In Argentinean estuaries, both species parasitize the cochliopid snail Heleobia australis as first intermediate host, the grapsid crabs Neohelice granulata and Cyrtograpsus angulatus as second intermediate hosts, and gulls as definitive hosts. Here, we describe the daughter sporocysts and cercariae of M. orensense and redescribe these stages for M. bonaerense. Sporocysts of M. orensense are shorter, with fewer developed cercariae than M. bonaerense. The cercariae of M. orensense have longer, larger, and more undulating cephalic glands than M. bonaerense. We redescribe metacercariae and adults of both species and compare them with the previous descriptions. Intestinal ceca length, vitellaria shape and extension, and egg size are the most relevant characteristics in metacercariae and adults for differentiating the species. Hence, the detailed morphological description and comparative analyses of morphometrics obtained from natural and experimental infections permit clear differentiation of M. orensense and M. bonaerense at each life stage.

We sampled 339 fecal samples, 296 intestines, and 82 lungs from 371 lynx hunted during the 2010–2011 season in Finland. The fecal samples were analyzed for endoparasites by a quantitative flotation method, and helminths from intestines were studied morphologically, while lungs were investigated for pulmonary parasites. From fecal samples, eggs and oocysts of at least 6 different endoparasite species were identified, with a mean of 1.5 (range 0–4) parasite species per host. In the intestines, at least 4 different helminth species were found, with the mean of 2.0 (range 1–4) species per infected host. The prevalence of eggs in feces and the prevalence of worms in intestines were 71% and 93% for Toxocara cati, 29% and 68% for Taenia spp., and 5% and 2% for Diphyllobothrium sp., respectively. Only eggs were detected for Capillaria sp. (46%) and Uncinaria sp. (0.6%) nematodes, and only adults were detected for Mesocestoides sp. cestodes (0.3%). Significant positive correlations were evident between the number of T. cati (r = 0.664; P = 0.01) and Diphyllobothrium sp. (r = 0.645; P = 0.01) eggs per gram of feces and adult worms detected in intestine. In addition to the metazoan parasites, protozoan Isospora sp. oocysts were also found (0.6%). Pulmonary samples were all negative for parasites. These data demonstrate that lynx commonly harbor various endoparasites, some of which are zoonotic.

Small amoeboid cells, believed to be the infectious stage of Ichthyophonus sp., were observed in the bolus (stomach contents) and tunica propria (stomach wall) of Pacific staghorn sculpins and rainbow trout shortly after they ingested Ichthyophonus sp.–infected tissues. By 24–48 hr post-exposure (PE) the parasite morphed from the classically reported multinucleate thick walled schizonts to 2 distinct cell types, i.e., a larger multinucleate amoeboid cell surrounded by a narrow translucent zone and a smaller spherical cell surrounded by a “halo” and resembling a small schizont. Both cell types also appeared in the tunica propria, indicating that they had recently penetrated the columnar epithelium of the stomach. No Ichthyophonus sp. pseudo-hyphae (“germination tubes”) were observed in the bolus or penetrating the stomach wall. Simultaneously, Ichthyophonus sp. was isolated in vitro from aortic blood, which was consistently positive from 6 to 144 hr PE, then only intermittently for the next 4 wk. Small PAS-positive cells observed in blood cultures grew into colonies consisting of non-septate tubules (pseudo-hyphae) terminating in multinucleated knob-like apices similar to those seen in organ explant cultures. Organ explants were culture positive every day; however, typical Ichthyophonus sp. schizonts were not observed histologically until 20–25 days PE. From 20 to 60 days PE, schizont diameter increased from ≤25 μm to ≥82 μm. Based on the data presented herein, we are confident that we have resolved the life cycle of Ichthyophonus sp. within the piscivorous host.

Experimental infections and field-collected lizards were used to investigate issues of transmission, host specificity, and seasonal occurrence in the nematode Cyrtosomum penneri (Cosmocercoidea: Atractidae). Anolis sagrei (87 males, 42 females) were captured from the Florida Southern College campus, Polk County, Florida, from October 2010 to September 2011, and 8,803 C. penneri were collected from their intestines. During the breeding season all sexually mature (SVL ≥34 mm) A. sagrei were infected, whereas juvenile lizards (SVL <34 mm) were never infected. Experimental infections, using A. sagrei, found that worms were transferred to new hosts venereally, but not during oral exposures. Mating trials confirmed that worms were consistently transferred between hosts during copulation under natural conditions. Experimental exposures found that land snails and crickets do not serve as transport or intermediate hosts, which supports the idea that C. penneri is transferred only during host copulation. Experimental infections to test host specificity in C. penneri successfully infected A. sagrei, Hemidactylus turcicus, and Sceloporus undulatus, but not Anolis carolinensis or Plestiodon inexpectatus. Overall, this is the first study to fully elucidate the life cycle of any atractid nematode, and we suggest a venereal route of transmission for all atractid worms that infect reptilian hosts. Our findings also have implications for the host's reproductive and behavioral biology, e.g., support for covert or satellite males in the A. sagrei mating system.

The migratory response of Echinostoma caproni to host feeding was examined in female ICR mice. Thirty-six mice were each infected with 20 metacercariae of E. caproni. Twenty-eight days post-infection, food, but not water, was withheld for 24 hr. Mice were haphazardly divided into 4 groups of 9, and each group received one of the following treatments: (1) 0.25 g glucose, (2) access to standard lab chow, (3) 0.5 ml saline, and (4) continued fasting. Three mice from each treatment group were killed 1, 2, and 4 hr post-treatment. The intestine of each mouse was removed, flash-frozen, and stored in a conventional freezer for later examination. Intestines were partially thawed, measured, and opened longitudinally, and the position of each worm, or worm cluster was measured. The intestine was divided into equal 5% segments based on the initial measurement and locations of worms, and worm clusters were recorded from the appropriate section of the intestine for analysis. There was no significant effect of treatment in the position of worms at 1 hr. There was a posterior shift in worm position in all treatment groups at 2 hr, except in the saline-treated mice; however, only worms in the glucose-fed mice were significantly posterior to the unfed controls. From 2 to 4 hr, there was a significant anterior movement of worms in both the glucose and chow-fed mice. The data strongly suggest that E. caproni responds to the initiation of gastric activity of the host by migrating anteriorly in the ileum. The specific stimulus for this migration is unknown.

Exposure to parasites is considered to be an important factor in the development of many diseases and histopathologies which are the result of the parasite–host interaction. The present study evaluated the impact of natural infection by larvae of Ortleppascaris sp. (Nematoda: Ascaridida) in the liver of the cane toad Rhinella marina (Linnaeus, 1758). Larvae were encysted in nodules delimited by collagenous fibers and fibroblasts or freely within the hepatic parenchyma, provoking a clear response from the host. The histological examination of the liver revealed viable larvae in a number of different developmental stages, as well as cysts filled with amorphous material and cell residues and surrounded by dense fibrotic tissue. The infection of the liver by these larvae induces a significant increase in the area occupied by melanomacrophages and a reduction or deficit in the vascularization of the liver, hypertrophy of the hepatocytes, vacuolar bodies, and cytoplasmatic granules. Focal concentrations of inflammatory infiltrates were observed enclosing the unencapsulated early-stage larvae. These results indicate that infection by Ortleppascaris sp. induces severe physiological problems and histopathological lesions in the liver of R. marina.

Acanthogyrus (Acanthosentis) barmeshoori n. sp. (Quadrigyridae) is described from the Persian tooth-carp, Aphanius farsicus Teimori, Esmaeili, and Reichenbacher, 2011 (Cyprinodontidae) in the Maharlu Lake basin, southern Iran. Aphanius farsicus is an endemic freshwater fish found in streams and springs that drain into Maharlu Lake, Shiraz, Iran. The new species is the smallest of all the 44 known species of the subgenus Acanthosentis Verma and Datta, 1929, measuring between 0.26 and 1.68 mm in length. It is further distinguished by having a short cylindrical proboscis with very long anterior hooks widely separated from very small hooks in 2 very close circles posteriorly (hook length ratio about 4:1). It is separated from 4 other species of Acanthosentis with similar proboscis armature but with less-extreme diversification of hook length. The new species is also distinguished in having anterior para-receptacle structures (PRS) and a similar posterior structure like those reported in only 1 other species of Acanthosentis from Japan. Proboscis receptacle is single walled with a large triangular cephalic ganglion. Testes are large, pre-equatorial, and Saefftigen's pouch is prominent. Fourteen to 25 circles of spines cover the anterior 50–70% of the trunk, but a few spines may be present at posterior end of trunk. This is the first species of Acanthosentis where SEM images, showing external morphological details, are provided. From a total of 357 fish specimens examined between July 2006 and June 2007, 173 specimens (48.5%) were infected with individuals of the new species. The prevalence of infection decreased with increasing fish size. The parasite was observed all year, with the highest abundance and intensity in May while the prevalence was highest in February. The prevalence of acanthocephalans decreased with increasing fish size. While most worms were recovered in fish within the length range of 18–29.9 mm, 1 of the longest parasites (1.68 mm long) was found in fish within the range of 30–33.9 mm long.

Histomonas meleagridis, a flagellated protozoan of the Order Trichomonadida, is the causative agent of blackhead disease in gallinaceous birds. Few genes have been identified in this organism; thus, little is known regarding the molecular basis for its metabolism, virulence, and antigenicity. To identify new genes, a cDNA library derived from a lab strain of H. meleagridis was sequenced and annotated. Data obtained from these experiments identified 3,425 H. meleagridis genes. Analysis of the data allowed the identification of 81 genes coding for putative hydrogenosomal proteins and was used to determine the codon usage frequency. Sequence information also identified bacteria that are cultured with H. meleagridis. Future analysis of these data should provide valuable molecular insights into H. meleagridis and provide the platform for molecular studies aimed at understanding the pathogenesis of blackhead disease.

The Socorro dove Zenaida graysoni, endemic to Socorro Island, was last reported in the wild in 1972. Fortunately, the species has been propagated in zoos in Europe and the United States, and plans are under way to re-introduce it to its native habitat. This will be the first known attempt to return a bird species extinct in the wild to its ancestral island. In order to assess the disease threats the Socorro dove may face, the avifauna of Socorro Island, with a specific focus on Socorro ground doves Columbina passerina socorroensis and mourning doves Zenaida macroura, as well as Socorro doves in captivity, were screened for blood parasites of the genera Plasmodium, Haemoproteus, Leucocytozoon, and Trypanosoma spp. We found Haemoproteus spp. in 17 (74%) of 23 Socorro ground doves, 23 (92%) of 25 mourning doves, and 3 (14%) of 21 northern mockingbirds; none of the other bird species showed infections. Here, we report the phylogenetic analysis of 19 distinct lineages of Haemoproteus spp. detected in birds of Socorro Island and compare their evolutionary relationships to parasites detected in the avifauna of the Galápagos Islands, continental Latin America, and Europe. Microscopic examination revealed 1 mourning dove infected with Plasmodium (Haemamoeba), thus underscoring the importance of using both PCR and microscopy when analyzing avian blood samples for hemosporidian parasites. The study confirms that the Socorro dove will most likely be exposed to Haemoproteus spp. that currently infect mourning doves and Socorro ground doves of Socorro Island. A monitoring program for both birds and vectors should be implemented to establish the prevalence of Plasmodium sp. and as a necessary conservation measure for critically endangered birds on the island.

Lungworms of the cosmopolitan genus Rhabdias are among the most common parasites of amphibians and squamate reptiles. The present study used experimental infections, field studies, and a molecular phylogeny to determine the host specificity of 6 Rhabdias spp. that infect snakes and anurans from North America. The molecular phylogeny suggests Rhabdias ranae from Nebraska and Mississippi may represent separate, cryptic species. In addition, the phylogeny strongly supports separate clades for anuran and snake lungworms. Field studies and experimental infections indicate that snake lungworms are generalist snake parasites; however, laboratory experiments also suggest that lizards can be infected under some environmental conditions. Lungworms from anurans were found not to infect salamanders or reptiles, in nature or in the laboratory; anuran lungworm species ranged from strict host specificity, e.g., R. ranae from Nebraska, to relative generalist, e.g., Rhabdias joaquinensis from Nebraska. Overall, host specificity for species of Rhabdias does not provide support for the evolution of progressive specialization over time. For most species of lungworms, host specificity in nature appears to be limited by both ecological and physiological factors, which vary between species and their hosts. Furthermore, some lungworms, e.g., Rhabdias bakeri from Missouri, appear to be tracking host resources instead of host phylogenies, an example of ecological fitting.

Between March 1989 and February 1994, 4 bald eagles (Haliaeetus leucocephalus) from various localities in Kansas were examined for coccidia. One (25%) of the bald eagles was found to be passing an undescribed species of Caryospora in its feces. Oocysts of Caryospora hanebrinki n. sp. are ellipsoidal to ovoidal with a bilayered wall and measure 48.1 × 42.1 μm with a shape index of 1.2. A micropyle, oocyst residuum, and polar granule were absent. Sporocysts are spheroidal, 24.8 μm wide. Stieda, substieda, and parastieda bodies were absent; a spheroidal sporocyst residuum is present; it measures 17.5 μm and is composed of many intact homogenous globules with a few dispersed in a loose spiral around the sporocysts. This is the first caryosporan documented from the bald eagle and is the largest known Caryospora from raptors.

A new nematode species, Philometra atlantica n. sp. (Philometridae), is described from male and female specimens found in the ovary of the Atlantic Spanish mackerel, Scomberomorus maculatus (Mitchill) (Scombridae, Perciformes), off the Atlantic coast of Florida and South Carolina. Based on light and scanning electron microscopy examination, the new species differs from most other gonad-infecting Philometra spp. in the length of spicules (111–126 μm), number and arrangement of genital papillae, and a U-shaped, dorsally interrupted caudal mound in the male. A unique feature among all gonad-infecting philometrids is the presence of 2 reflexed dorsal barbs on the distal end of the gubernaculum. From a few congeneric, gonad-infecting species with unknown males, it can be distinguished by some morphological and biometrical features found in gravid females (body length, length of first-stage larvae or esophagus, structure of caudal end) and by the host type (fish family) and geographical distribution. Philometra atlantica is the fourth valid gonad-infecting species of Philometra reported from fishes of the family Scombridae.

A new nematode species, Rhabdochona (Rhabdochona) hypsibarbi n. sp. (Rhabdochonidae), is described from the intestine of the freshwater cyprinid fish Hypsibarbus wetmorei (Smith) in the Mekong River, Nakhon Phanom Province, northeast Thailand. It is mainly characterized by medium-sized, bifurcate deirids, the presence of 14 anterior prostomal teeth and distinct basal teeth, length ratio of the muscular and glandular portions of esophagus (1:6–9), length of the left spicule (669–774 μm), absence of a dorsal barb on the right spicule, length ratio of spicules (1:4.9–6.0), arrangement of genital papillae, and smooth eggs without filaments or swellings. In addition, specifically unidentified fourth-stage larvae of Rhabdochona (Rhabdochona) sp., morphologically similar to R. hypsibarbi, were recorded from the red-tailed tinfoil Barbonymus altus (Günther) (Cyprinidae) in the Mekong River, Nakhon Phanom Province, northeast Thailand. Rhabdochona hypsibarbi is the fourth nominal species of Rhabdochona Railliet, 1916 reported from fishes in Thailand and the second species of the nominotypical subgenus found in this country.

Rhabdias himalayanus n. sp. from the lungs of Duttaphrynus himalayanus and Rhabdias dehradunensis n. sp. from the lungs of Nanorana minica from Dehradun, India are described and figured. Of the 3 previously described Indian species, Rhabdias himalayanus n. sp. is most similar to Rhabdias shortii in having a cylindrical corpus, inflated cuticle, and conical tail; it differs from R. shortii in having greater body measurements, longer esophagus, larger eggs, and a different pattern of cuticle inflation at the vulva and tail region. Rhabdias dehradunensis n. sp. is most similar to Rhabdias bulbicauda in that both possess a swollen posterior end; it differs from R. bulbicauda by having a subterminal anus, a prominent tail, and a postequatorial vulva.

Microscopic and molecular procedures are used to describe a new myxosporean species, Chloromyxum clavatum n. sp., infecting the cartilaginous fish Raja clavata Linnaeus, 1758 (Chondrichthyes: Rajidae), collected from the northwest Atlantic coast of Portugal. Young plasmodia and mature spores were found floating free in the gall bladder of R. clavata. Spores were spherical to subspherical with a pointed anterior end, measuring14.4 ± 0.5 μm (n = 25) in length, 11.9 ± 0.5 μm (n = 25) in width, and 9.4 ± 0.5 μm (n = 15) in thickness. The spore's wall was composed of 2 equally sized valves, each displaying 6–8 elevated surface ridges and a bundle of several tapering caudal filaments attached to the basal portion. Spores contained 4 pyriform equally sized polar capsules (5.5 ± 0.4 μm × 2.9 ± 0.5 μm) (n = 25), each possessing an obliquely arranged isofilar polar filament coiled in 7–8 coils. Morphological data, host specificity, tissue tropism, and molecular analysis of the SSU rDNA gene identify this parasite as a new species of Chloromyxum. Neighbor-joining and maximum likelihood further reveal the parasite clustering with other species of Chloromyxum infecting the gall bladder of marine cartilaginous fish to form a clade positioned at the base of the freshwater clade, therefore constituting an exception to the major division of the class Myxosporea into the freshwater and marine clades, while supporting the existence of a correlation between tissue tropism and myxosporean phylogeny.

Eggs and larvae of Huffmanela oleumimica n. sp. infect red snapper, Lutjanus campechanus (Poey, 1860), were collected from the Texas–Louisiana Shelf (28°16′36.58″N, 93°03′51.08″W) and are herein described using light and scanning electron microscopy. Eggs in skin comprised fields (1–5 × 1–12 mm; 250 eggs/mm²) of variously oriented eggs deposited in dense patches or in scribble-like tracks. Eggs had clear (larvae indistinct, principally vitelline material), amber (developing larvae present) or brown (fully developed larvae present; little, or no, vitelline material) shells and measured 46–54 μm (x̄ = 50; SD ± 1.6; n = 213) long, 23–33 (27 ± 1.4; 213) wide, 2–3 (3 ± 0.5; 213) in eggshell thickness, 18–25 (21 ± 1.1; 213) in vitelline mass width, and 36–42 (39 ± 1.1; 213) in vitelline mass length with protruding polar plugs 5–9 (7 ± 0.6; 213) long and 5–8 (6 ± 0.5; 213) wide. Fully developed larvae were 160–201 (176 ± 7.9) long and 7–8 (7 ± 0.5) wide, had transverse cuticular ridges, and were emerging from some eggs within and beneath epidermis. The new species differs from its congeners by having eggs <65 μm in total length and that have a brown eggshell when fully developed, an envelope throughout development, and irregularly-dispersed eggshell spines plus a larva >110 μm long with transverse cuticular ridges. The eggs lack a spindle-shaped envelope, polar filaments, and eggshell ridges. This is the first report of a species of Huffmanela from a snapper (Lutjanidae) or from the Gulf of Mexico. A table of egg and larval characteristics, hosts, and localities for Huffmanela spp. is provided.

Angiostrongylus felineus n. sp. (Nematoda, Metastrongyloidea), parasitic in Puma (Herpailurus) yagouaroundi (É. Geoffroy, 1803) (Carnivora, Felidae) from the municipality of Juiz de Fora, Minas Gerais state, Brazil, is described and illustrated herein. Angiostrongylus felineus n. sp. differs from all congeneric species by having the anterior extremity with accentuated cuticular expansion and by smaller size of spicules. This study describes for the first time a species of Angiostrongylus in a wild Felidae in Brazil.

The infective larvae of Teladorsagia circumcincta have a protective sheath that is lost soon after they reach the rumen of the sheep (the definitive host). Incubation in vitro with 50 mM imidazole caused more than 75% of L₃ T. circumcincta to begin exsheathing within 2 hr. The initiation of exsheathing was less likely at pH 6.2 than at pH 7.8. The apparent pK_a of this process was 7.08, similar to that for the conversion of imidazolium to imidazole. Both the extent and the initial rate of exsheathing initiation increased with imidazole concentration (the apparent K_½ was about 50 mM). The initial rate of exsheathing initiation was stimulated by lactose and maltose, but not by some other carbohydrates, and by propylamine and imidazole acetic acid, but not by histidine.

Because of outbreaks of cryptosporidiosis in humans, some Cryptosporidium spp. have become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by Cryptosporidium suis, a fraction may be comprised of other Cryptosporidium species infectious to humans such as Cryptosporidium parvum. To better understand the survival dynamics of Cryptosporidium spp., oocysts associated with swine operations, 2 experiments were performed to determine die-off rates of C. parvum oocysts in a swine waste lagoon (2009 and 2010) and its spray field (2010 and 2011). Sentinel chambers containing a lagoon effluent suspension of C. parvum oocysts were submerged in the lagoon, and triplicate chambers were removed over time; oocysts were extracted and assayed for viability. For comparative purposes, inactivation rates of Ascaris suum eggs contained in sentinel chambers were also determined. For 2 spray field experiments, air-dried and sieved surface soil was placed in sentinel chambers, hydrated, and inoculated with a lagoon effluent suspension of C. parvum oocysts. Sentinel chambers and control oocysts in PBS contained in microcentrifuge tubes were buried 1.5 cm below the soil surface in 3 blocks. Triplicate chambers and controls were removed over time; oocysts were extracted and assayed for viability. Based on the first order decay equation, days to reach 99% die-off (T₉₉) were determined. T₉₉-values determined for the 2 lagoon experiments were 13.1 and 20.1 wk, respectively. A T₉₉-value for C. parvum in the spray field was significantly longer at 38.0 wk than the control oocysts in PBS at 29.0 wk. The waste lagoon and spray field system of manure management at this large-scale farrowing operation appeared to reduce the load of C. parvum oocysts before they can be hydrologically transported off the operation and reduces their likelihood of contaminating surface waters and threatening public health.

The aim of the present study was to investigate antibodies to Toxoplasma gondii in the serum of mules and donkeys bred in the northeast of Brazil. In total, 483 samples were used (395 mules and 88 donkeys) from 4 states (Pernambuco, Rio Grande do Norte, Paraíba, and Sergipe). The indirect immunofluorescence reaction (IFI) technique was used to investigate antibodies to T. gondii with a cut-off point of 64. Positive frequencies of 23.8% and 43.2% were recorded for mules and donkeys, respectively. The state of Pernambuco had the highest prevalence of positive samples (29%) with statistically significant differences for species (P < 0.001) and state (P = 0.048). This is the first study of antibodies to T. gondii in mules and donkeys in these 4 states of the northeastern region of Brazil and serves as a warning to health authorities regarding the risks of ingesting equine meat.

The tick Ixodes brunneus Koch is a rare species occurring primarily in North America, where it feeds on many species of passeriform birds. Virtually nothing is known about the questing activity of this tick, although adults often stand with their front legs straight up, suggesting that they quest from a horizontal position. The present study analyzed I. brunneus questing behavior based on field data from drag cloth collections in northern Mississippi, as well as observational laboratory data from 10 I. brunneus ticks released into an experimental “questing apparatus.” Ten ticks of a related species, I. scapularis Say, were used for comparison, and there were 3 replications each trial. Eight I. brunneus adults were collected along a nature trail in a northern Mississippi park during 20 total swaths with a drag cloth over a 2-day period (each time 5 swaths in the middle of the trail, with little or no vegetation; and 5 swaths along the edge of the trail, with taller vegetation). All 8 ticks were collected in the middle of the trail in vegetation no taller than 40 mm. In the laboratory experiment, the majority (>70%) of ticks of both species made no attempt to climb the metal or wood artificial stems, but instead they crawled around on the substratum. In 8/30 instances, I. brunneus climbed metal artificial stems to various heights as opposed to 4/30 instances for I. scapularis. Sometimes, ticks of both species seemed to quest at the base of both types of artificial stems. The mean height for questing by I. scapularis on metal stems was 38.2 mm as opposed to 31.8 mm for I. brunneus. Although the mean height was slightly higher for I. scapularis compared with I. brunneus, there was no statistical difference in questing heights observed between the 2 species. Ixodes brunneus and I. scapularis climbed wooden artificial stems in only 2/30 instances for each tick species, again with no statistical difference in questing heights between species. The field observations suggest that I. brunneus quests at ground level; however, laboratory simulations revealed that although the average height climbed was below 40 mm, they sometimes climb artificial stems to greater heights (approximately 50 mm in 4 instances).

Babesiosis is a tick-borne protozoan disease affecting many mammalian species worldwide, caused by the intraerythrocytic multiplication of Babesia spp. The present study aimed to detect the presence of Babesia sp. in 13 American mink from Hokkaido, Japan. One of 13 animals was positive, as indicated by nested PCR targeting the 18S ribosomal RNA (SSU rDNA) and subunit 7 (eta) of the chaperonin-containing t-complex polypeptide 1 (CCT7) genes from species of Babesia and Theileria. Sequencing of the PCR product of SSU rDNA revealed 99% homology to the isolates of Babesia sp. SAP#131 found in raccoons in Hokkaido, whereas that of the CCT7 gene showed 80% homology to the isolates of Babesia gibsoni in dogs as determined by BLAST analysis. We refer to the cognate sequence as Babesia sp. NV-1. Phylogenetic analyses of SSU rDNA and CCT7 genes from Babesia sp. NV-1 revealed them to be most closely related to the Babesia sp. SAP#131 from a raccoon in Hokkaido and to canine B. gibsoni, respectively. Here, we provide the first molecular evidence of the Babesia sp. NV-1 parasite in feral American mink (Neovison vison) in Hokkaido, Japan.

In view of the interest in genotype-specific pathogenesis in Giardia duodenalis, the aim of the present study was to examine the effects of infection with different, or mixed, G. duodenalis assemblages on the integrity of human intestinal epithelia. To that end, human epithelial cells (HCT-8) were cultured and exposed to different G. duodenalis assemblages (A, B, and E) or a combination of these assemblages. Epithelial disruption and apoptosis were evaluated by fluorescent microscopy and apoptotic oligonucleosome quantification. The results indicate that infection with trophozoites disrupts epithelial tight junctions and induces varying degrees of enterocyte apoptosis, depending on the infecting assemblage. All disruptions were caspase-3 dependent and were more pronounced when caused by a non-host specific assemblage. Furthermore, infections by isolates in combination with isolates from another assemblage enhanced the epithelial disruption and apoptosis. Further studies in vitro and in vivo are required to confirm the mechanisms of enhanced pathogenicity of mixed or non-host specific (or both) G. duodenalis infections. Findings in the present study point to the potential pathogenic importance of intra-species polyparasitism in giardiasis.

Leucocytozoon spp. infections have been rarely studied in Neotropical countries. The apparently low prevalence of these parasites compared to the Nearctic regions suggests the absence of competent vectors; however, a 21.3% overall prevalence has recently been reported in non-migratory birds from the páramo region of Chingaza National Natural Park (NNP), where Turdus fuscater (Great Thrush) is the species most frequently infected by these parasites. The present study provides the descriptions of the Leucocytozoon spp. detected in Great Thrushes trapped in Chingaza NNP. The parasites were confirmed by microscopic examination and PCR of blood, histopathology was also done. Leucocytozoon dubreuili and L. fringillinarum gametocytes were observed in blood smears. The corresponding cytochrome b (cyt b) lineages obtained of L. fringillinarum were closely related to lineages previously found in individuals infecting turdiid species sampled elsewhere. This is one of the few reports analyzing Leucocytozoon spp. infections in resident birds from a Neotropical country.

The bloodstream form of Trypanosoma brucei acquires iron from transferrin by receptor-mediated endocytosis. However, it is unknown how procyclic forms that cannot bind transferrin acquire iron. Here, we show that the procyclic form of T. brucei efficiently takes up iron from ferric complexes via a reductive mechanism and that iron obtained using this mechanism is transported to, and used in, the mitochondria. The affinity of the transport system is comparable to that of Saccharomyces cerevisiae, with an apparent K_m of 0.85 μM.

Neospora caninum is an important cause of bovine abortion worldwide for which dogs are the definitive host. The present study was aimed at investigating the exposure to N. caninum infection based on lifestyle categories of dogs from southern Romania. For this purpose, randomly selected rural and urban dogs were examined for fecal N. caninum-like oocysts and were serologically tested for the presence of anti-N. caninum IgG antibodies. Of the 386 dog fecal samples, N. caninum-like oocysts were found in 19 (4.9%; 95% CI = 2.89–7.59) as follows: rural guard dogs (4/41; 9.8%), cattle farm dogs (6/118; 5.1%), and stray dogs (9/192; 4.7%) (P > 0.05). None of the 35 urban guard dogs was positive. Serum samples (n = 84) from all of the 19 N. caninum-like oocysts-positive dogs and another 65 randomly selected canines (15 cattle farm dogs, 21 rural guard dogs, and 29 strays) were tested by indirect fluorescent antibody test (IFAT). None of these dogs exhibited any symptoms of clinical neosporosis. However, IgG antibodies against N. caninum were detected in 17/84 (20.2%) (P < 0.05) serum samples. The highest prevalence was registered in cattle farm dogs (38.1%) followed by strays (18.4%) and rural guard dogs (8.0%). The seropositivity to N. caninum increased significantly with age (P < 0.05), reaching 66.7% in dogs >10 yr of age, suggesting post-natal exposure to N. caninum is the predominant mechanism of N. caninum recruitment.

Neospora caninum is a major cause of bovine abortion worldwide. A serological survey was carried out to determine the seroprevalence of exposure to N. caninum in dairy cattle based on age and breed from Punjab and Sindh provinces, Pakistan. Serum samples from 641 animals from 12 herds from Punjab (n = 7) and Sindh (n = 5) provinces were tested for antibodies against N. caninum using a commercially available competitive enzyme-linked immunosorbent assay. Positive reactions to N. caninum were seen in 277 (43%) of the 641 of the samples. Seropositive animals were present in all 12 herds. Animals over 2 yr of age (47%) and crossbreds (55%) were more likely to be seropositive than the other cattle examined. These results indicate that N. caninum infection is widespread among dairy cattle in Pakistan.

The protozoan parasite Toxoplasma gondii is globally distributed, with considerable local variation in prevalence based on behavioral and environmental factors. To assess prevalence and estimate risk in Mali, we conducted a survey of 760 serum samples previously collected for malaria studies. A modified agglutination test detected antibodies in ∼27% of the adult population, with no significant differences between men and women, or between urban and rural study sites. In the village of Kolle, seroprevalence rose from 0% in infants (<1 yr, but after weaning of maternal immunoglobulin G) to 0.8% (1–5 yr), 2.7% (6–10), 11.3% (11–15), and 26.8% (>15); differences between the <10-, 11–15-, and >15-yr age groups were highly significant (P ≤ 0.01). We also observed an increase in anti–T. gondii antibody titers with age. Modeling the observed age distribution suggests a seroconversion rate of ∼1%/yr, indicating that congenital toxoplasmosis may be an under-appreciated public health concern in Mali.

To better define the strains and species of Hepatozoon that infect coyotes in the south-central United States, whole blood and muscle samples were collected from 44 coyotes from 6 locations in Oklahoma and Texas. Samples were evaluated by a nested polymerase chain reaction (PCR) using primers amplifying a variable region of the apicomplexan 18S rRNA gene as well as histopathology (muscle only) for presence of tissue cysts. Hepatozoon spp. infections were identified in 79.5% (35/44) of coyotes tested including 27 of 44 (61.4%) whole blood samples and 17 of 44 (38.6%) muscle samples tested by PCR and 23 of 44 (52.3%) muscle samples evaluated by histological examination. Analysis revealed 19 distinct sequences comprising 3 major clusters of Hepatozoon spp., i.e., 1 most closely related to Hepatozoon americanum, another most closely related to Hepatozoon canis, and the third an intermediate between the 2 groups. The diversity of Hepatozoon spp. in wild canids appears greater than previously recognized and warrants further investigation.

We have described the development of a 2-step nested PCR protocol based on the characterization of the 70-kDa heat shock protein (HSP70) gene for rapid detection of the human-pathogenic Cyclospora cayetanensis parasite. We tested and validated these newly designed primer sets by PCR amplification followed by nucleotide sequencing of PCR-amplified HSP70 fragments belonging to 16 human C. cayetanensis isolates from 3 different endemic regions that include Nepal, Mexico, and Peru. No genetic polymorphism was observed among the isolates at the characterized regions of the HSP70 locus. This newly developed HSP70 gene-based nested PCR protocol provides another useful genetic marker for the rapid detection of C. cayetanensis in the future.

The objective of this study was to estimate the seroprevalence of anti–Toxoplasma gondii antibodies and DNA of women with spontaneous abortions in 2 hospitals located in Yucatan, Mexico. Between June 2008 and May 2009, blood samples were taken from 100 women with spontaneous abortion attending the Ticul City Communitarian Hospital and the Merida Mother–Kid Hospital. The sera were tested for anti–T. gondii IgG and IgM antibodies. Blood samples (5 ml with anticlotting agent) were also used for polymerase chain reaction (PCR) assay, to detect T. gondii DNA. Forty-two of the 100 samples were negative. Of the positive samples (n = 58), 32 were positive to IgG, 2 to IgM, 5 to IgG and IgM, 6 to IgG and PCR, 1 to IgM and PCR, and 12 to IgG, IgM, and PCR. Accordingly, 55% of the women were seropositive to at least IgG, 20% to at least IgM, and 19% via PCR. Differences between hospitals were significant (P < 0.05) only for IgM. The risk of infection (IgM positive) was 2.85 (odds ratio [OR] 95%, confidence interval [CI]; 1.03–7.87) times greater in women patients at the Merida Mother–Kid Hospital, than those at the Ticul Communitarian Hospital. More studies are needed to evaluate the impact of this disease and to establish strategies to follow in order to reduce congenital toxoplasmosis in the populations at risk.

Prevalence is one of the few estimates that rarely are reported with an appropriate measure of error in the parasitological literature. A minimum sample size recommendation of 15 samples, based on the relationship between sample size and standard error, likely has led to a false degree of confidence because of the nonlinear relationship between standard error and “true” 95% confidence intervals (as determined by Monte Carlo simulation or integration of the Bayesian posterior). Given that 95% confidence intervals for proportions are influenced by both sample size and the actual estimate of the proportion, there is no “gold standard” sample size beyond which estimates of binomial proportions can be considered “reliable.” This necessitates the reporting of confidence interval estimates that have been shown to be conservative, such as the Clopper-Pearson estimate, or robust, such as the Wilson score approximation, or the computationally intensive integration of the Bayesian posterior.