In vitro cultivation of adult digeneans can benefit research on their biology and contribute to the development of new drugs and vaccines. Successful in vitro growth of excysted metacercariae into adults capable of producing embryonated eggs typically requires that the worms be inseminated. The goal of the study was to develop an in vitro insemination procedure for the progenetic microphallid digenean Gynaecotyla adunca. To do so, we determined the length of time needed for in vitro sperm development in excysted metacercariae and whether the adult worms could self-inseminate in the absence of conspecifics. We also examined the effect of different culture vessels, worm densities, incubation temperatures, length of time incubated with conspecifics, and different pH levels on the percentage of worms inseminated. We found that sperm maturation time for G. adunca was 8–10 hr postexcystment. In the absence of conspecifics, the parasite did not self-inseminate. We observed the highest percentage of inseminated worms when 50 excysted metacercariae were incubated at 37 C for 48 hr in 15-ml conical-bottom tubes containing pH 7 Hank's balanced salt solution. Furthermore, freshly excysted worms incubated in these conditions and then transferred to culture in Dulbecco's modified Eagle medium/F-12 medium and horse serum deposited normal-shaped, embryonated eggs. Our findings provide the basis for a straightforward, reproducible procedure that permits the in vitro insemination of the parasite G. adunca and should be applicable to other progenetic digeneans.

Complex societies where individuals exhibit division of labor with physical polymorphism, behavioral specialization, and caste formation have evolved several times throughout the animal kingdom. Recently, such complex sociality has been recognized in digenean trematodes; evidence is limited to 6 marine species. Hence, the extent to which a soldier caste is present throughout the Trematoda is sparsely documented, and there are no studies detailing the structure of a species lacking such a social structure. Here we examine colony structure for an additional 5 echinostomoid species, 4 of which infect the marine snail Cerithidea californica and 1 (Echinostoma liei) that infects the freshwater snail Biomphalaria glabrata. For all species, we present redia morphology (pharynx and body size) and the distribution of individuals of different castes throughout the snail body. When morphological evidence indicated the presence of a soldier caste, we assessed behavior by measuring attack rates of the different morphs toward heterospecific trematodes. Our findings indicate that each of the 4 species from C. californica have a permanent soldier caste while E. liei does not. The observed intra- and inter-specific variation of caste structure for those species with soldiers, and the documentation of colony structure for a species explicitly lacking permanent soldiers, emphasizes the diverse nature of trematode sociality and the promise of the group to permit comparative investigations of the evolution and ecology of sociality.

This study describes the effect of increasing exposure dose on Ichthyophonus prevalence and infection intensity in experimentally infected rainbow trout, Oncorhynchus mykiss. Specific-pathogen free trout were exposed per os to increasing numbers of Ichthyophonus schizonts obtained from naturally infected donor fish, then sampled after 30 and 60 days post-exposure. Both in vitro explant culture and histology revealed that as the number of schizonts per dose increased there was a proportionate increase in the number of infected fish, as well as an increase in the number of infected organs; parasite density in individual infected organs also increased with dose. Explant culture revealed that all fish exposed to the highest dose (≥2,080 schizonts) became infected, while only 67% of those exposed to the intermediate dose (1,040–1,153 schizonts) were Ichthyophonus-positive after 60 days; Ichthyophonus was not detected in fish exposed to the 2 lowest doses (≤280 schizonts). Histologic examination of individual infected organs also revealed increasing infection prevalence and parasite density in response to exposure to increasing numbers of Ichthyophonus schizonts.

The Richelieu River, Quebec, is a highly-regulated waterway subject to numerous anthropogenic influences from municipal effluents and agricultural activities. Parasite communities in 234 spottail shiners (Notropis hudsonius) were examined from 4 localities in late spring 2003 and 2004. Component species richness varied between 15 and 18 species in 2003 but declined to 9 to 13 in 2004, while total parasite abundance was consistently lower in 2003. Parasite component community similarity among localities could not be directly linked to available upstream water quality measurements or anthropogenic activity and was best explained by precipitation. Total precipitation in May 2003 was approximately 40% more than in May 2004, presumably altering patterns of runoff, river flow rates, and water quality. This study suggests that fish parasite species composition and richness in the Richelieu River are influenced by environmental parameters which in turn ultimately are driven by a combination of climatic conditions and anthropogenic activities in the watershed.

The course of infection of Echinostoma caproni was followed in female ICR mice, a permissive laboratory host, from infection to natural termination. Twenty-one mice were infected with 20 metacercariae via oral intubation and housed 3 per cage. Three mice from a randomly selected cage were necropsied at 1 mo intervals. A second group of 15 mice was infected approximately 1 yr later to replace mice negative at necropsy in the first group. Mice in the second group were examined weekly for the presence of eggs in the feces. Mice negative for eggs on consecutive days were killed and necropsied. The location of individual worms and worm clusters were located in 20 segments of the small intestine. Adult worms were killed and fixed in hot formalin, stained, and prepared as whole mounts. Standard measurements were taken using a compound microscope fitted with an ocular micrometer. The infection spontaneously resolved in 10 mice from 7 to 32 wk PI, indicating the host response is highly variable and extending the maximum recorded length of E. caproni infections in ICR mice to 31 wk. A moribund worm was found in the feces of an animal that continued to pass eggs for an additional 2 mo indicating individual variation in worm responses. Worms located preferentially in the ileum (segments 11–13) during the first 3 mo of the infection but shifted to the jejunum (segments 8–9) during weeks 4–6. Morphologically, worms of different ages clustered together in multivariate space, with substantial overlap between the 3- and 4-mo-old infrapopulations and between the 5- and 6-mo-old infrapopulations. Muscular structures increased in size throughout the experiment, while the gonads increased in size for the first 3 mo and then declined during the last 3 mo. The relationship between E. caproni and ICR mice is more nuanced than previously reported. The reduction in gonad size and the shift from the ileum to the jejunum in the last 3 mo likely are related. These changes might be attributable to a localized immune response by mice to E. caproni that results in the ileum becoming less hospitable and a resultant relocation of the worms to a less favorable location in the jejunum.

Schistocephalus solidus plerocercoids commonly infect three-spined stickleback Gasterosteus aculeatus populations in brackish and freshwaters, but infections are typically absent from marine populations. Here we provide an experimental test of the salinity tolerance of S. solidus eggs, to determine the role of salinity in limiting the distribution of infection in coastal zones. We find that S. solidus eggs, derived from the in vitro culture of 3 different plerocercoids, developed normally in salinities of up to 12.5‰, but above this egg viability dropped rapidly, and no egg hatching was observed at salinities above 20‰. Our results are consistent with the distribution of infections in natural stickleback populations and add resolution to previous descriptive observations on salinity tolerance in S. solidus. They also demonstrate that S. solidus presents a novel disease challenge to marine populations of three-spined sticklebacks entering brackish and freshwater environments.

Helminths often demonstrate preferential site selection in which a parasite will only occur in 1 microhabitat or a restricted portion of its fundamental niche within its host. However, factors responsible for helminth site specificity are poorly understood, and very little is known about how these factors vary among multiple host species. Some helminths, such as Halipegus occidualis, have been reported from different habitats (stomach or under the tongue) within multiple anuran host species, suggesting that the site selected varies within anuran species. This study examined the site selection by H. occidualis in 7 definitive anuran host species using experimental infections. Then, the site fidelity of H. occidualis was further tested by transplanting worms from under the tongue to the stomach and vice versa in different anuran host combinations, and the movement of worms was recorded. Halipegus occidualis individuals occupied the habitat under the tongue in 6 of 7 anuran species. However, worms always occupied the stomach of American bullfrogs (Lithobates catesbeianus) and were never found under the tongue or in the mouth of these hosts. More importantly, all worms remained in the original habitat when transplanted from the stomach to the stomach or the buccal cavity to the buccal cavity within another individual of the same amphibian species. However, when worms were transplanted from the stomach to the buccal cavity or vice versa in the same host species, the worms always migrated back to the original habitat. The main contribution of this study is that it experimentally documented the variability in the site fidelity of H. occidualis within multiple definitive host species and determined that site fidelity is not as strongly conserved in this genus as suggested previously. Additionally, this work suggests that the variation in site selection in different host species could lead to speciation of the parasites.

Anaplasma marginale is the causative agent for bovine anaplasmosis (BA) and Ehrlichia ruminantium is the causative agent for heartwater, 2 devastating diseases of cattle. BA is common in the United States and frequently reported in western Tennessee cattle; however, cases of heartwater are not yet established in the continental United States. Because both pathogens are transmitted via the bites of infected ticks, the objective of this study was to survey cattle and pastures for ticks and for each pathogen. University of Tennessee AgResearch has 7 research and education centers (REC) located throughout the state at which they manage cattle. Ticks were collected from selected cattle (every fourth to sixth animal) and pastures (via dragging) associated with the herd from each REC during the summer of 2013. A total of 512 ticks were collected from cattle (n = 386) and pastures (n = 126) and were PCR-screened for Anaplasma and Ehrlichia using genus-specific primers. Collections consisted of 398 (77.7%) Amblyomma americanum, 84 (16.4%) Amblyomma maculatum, and 30 (5.9%) Dermacentor variabilis. Ticks were not recovered from pastures or cattle east of the Tennessee Plateau. The North American vectors for An. marginale and E. ruminantium were identified (D. variabilis and A. maculatum, respectively), but neither pathogen was recovered. A large proportion of ticks were collected from cattle and, of these, a majority were attached to their host (compared to questing on their host or engorged on the host). Four A. americanum were positive for Ehrlichia spp. (Ehrlichia ewingii, Ehrlichia chaffeensis, and Panola Mountain Ehrlichia), all in western Tennessee. With the identification of a few Ehrlichia infections in cattle-associated ticks and current A. marginale rates in Tennessee beef cattle nearing 11%, additional research is needed to establish baseline tick, Anaplasma, and Ehrlichia data for future management studies.

Rhinosporidium seeberi is an uncultivated Ichthyosporean infecting animals, including humans. Recent studies suggested R. seeberi undergoes synchronized nuclear division without cytokinesis. We used confocal microscopy to investigate R. seeberi nuclear division cycles in formalin-fixed tissues stained with DAPI and phalloidin. We report that R. seeberi nuclei in juvenile and intermediary sporangia synchronously divided without cytokinesis. Intermediary sporangia display numerous 3–4 μm nuclei at different mitotic stages as well as a thick inner layer with strong affinity for phalloidin. Mature sporangia showed numerous 5–12 μm cell-walled endospores, each containing a 2–4 μm in diameter nucleus. Phalloidin did not bind to the inner layers of mature sporangia or endospores. The development of a “germinative zone” in the inner layer of mature sporangia containing hundreds of nuclei was also confirmed. This study establishes that during the R. seeberi life cycle synchronous nuclear divisions without cytokinesis takes place, resulting in the formation of thousands of nuclei. Cytokinesis, on the other hand, is a 1-time event and occurs in the latest stages of intermediate sporangia, after the formation of thousands of nuclei and just before mature sporangia development.

Giardia duodenalis is considered the most common intestinal parasite in humans worldwide. Children are especially affected, with more severe consequences than adults. The present study was designed to determine the distribution of assemblages A and B Giardia infection in children and adults, with the use of light microscopy and polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) as diagnostic procedures, and to investigate its associations with clinical and epidemiological data collected from children and adult groups. This cross-sectional study was conducted from October 2012 to October 2013 by collecting fecal samples from 200 children and 200 adults complaining of diarrhea. Samples were subjected to parasitological examination by direct wet smear and formol-ether methods. Genotyping of G. doudenalis samples was conducted by PCR-RFLP analysis. Giardia duodenalis infection caused by assemblages A and B was identified in 60 samples, 34 from children and 26 from adults. Assemblage B was detected in 38 patients (63.34%), and assemblage A was detected in 22 patients (36.66%). Assemblage A was significantly more frequent in children with age range 2–8 yr, and assemblage B was higher in children with age range 6–16 yr old. Diarrhea frequency/day and recurrences per month affected patients infected with assemblage A (P value < 0.001) more frequently. Children infected with assemblage A presented significantly more severe diarrhea and dehydration than those infected with assemblage B (P value < 0.001). Although both Giardia assemblages A and B were identified in children and adults, assemblage A infected younger children more frequently and was more closely related to severe clinical manifestations than assemblage B.

We detail gross and histopathological changes associated with infection by the eggs, larvae, and cuticular remnants of Unionicola sp. in the mantle, gill, and visceral mass of 25 Alabama creekmussels, Strophitus connasaugaensis, collected during May 2010 through July 2012 from 2 Alabama streams. A multitude (estimated mean intensity >100) of mite eggs and larvae typically infected mantle, gill, and visceral mass integument. Pathology associated with eggs (prevalence = 0.57) and larvae (prevalence = 0.39) typically consisted of localized distension of the infection site; a host response to these infections was indeterminate. However, larval mites embedded in suprabranchial connective tissues were typically encapsulated (prevalence = 0.89). Mite remnants (prevalence = 0.5) occurred in mantle, gill, visceral mass integument, foot, heart, pericardial gland, intestinal lamina propria, and were typically encapsulated. We speculate that S. connasaugaensis clears some infections but is recolonized by autoinfection or horizontal dispersal of mites in the stream. Noteworthy is that high-intensity infections seemingly do not markedly impact the histological picture of mussel tissues, indicating that mites are relatively benign symbionts that are of little concern to mussels under normal environmental conditions.

The phylogenetic relationships of Cucullanidae were explored using near-complete sequences of the 18S rDNA (rRNA gene). Sequences (1,750–1,760 bp) were obtained from 7 species of Cucullanidae belonging to 3 genera, Cucullanus (2 spp.), Dichelyne (2 spp.), Truttaedacnitis (3 spp.), and 1 species of Quimperiidae (Paraseuratum sp.). These sequences were aligned with those of 128 other nematode species available in GenBank, including 3 other cucullanids (Dichelyne mexicanus, Cucullanus robustus, and Cucullanus baylisi) and 2 non-cucullanid seuratoids (Paraquimperia africana, and Linstowinema sp.). Bayesian (BPP) and maximum likelihood (ML) analyses of 2 different datasets strongly supported a monophyletic Cucullanidae. Bayesian analysis placed this family as the sister group to a clade containing species of Diplogasterida, Strongylida, Rhabditida, and Tylenchida with very strong support. Neither BPP nor ML analyses recovered a close relationship of Cucullanidae to Ascaridida. None of the 3 non-cucullanid seuratoid species were sister to Cucullanidae, nor did they form a monophyletic group of their own, which questions the monophyly of Seuratoidea and the relationships among species within this superfamily. The 3 genera of cucullanids were also not monophyletic, although morphologically similar species such as the 2 species of Cucullanus from Neotropical catfishes and 2 species of Dichelyne from Nearctic ictalurid catfishes were sister taxa with strong support. The results were ambiguous with respect to the relationship of 2 Truttaedacnitis spp. in Nearctic freshwater fishes but do not support Truttaedacnitis heterodonti, a parasite of heterodontid sharks, as belonging to this genus. The study shows that all aspects of the conventional classification of Seuratoidea and its taxa should be scrutinized by even more extensive sampling across hosts and habitats.

Two species of digeneans belonging in Homalometron are described from Nearctic freshwater fundulid fishes: Homalometron robisoni n. sp. is described from the Blackstripe Topminnow, Fundulus notatus, from Oklahoma and Homalometron frocioneae n. sp. is described from the Banded Killifish, Fundulus diaphanus, from New York. Homalometron robisoni n. sp. differs from all congeners by having vitelline follicles that extend into the forebody, a feature that necessitates altering the generic diagnosis for the genus. Homalometron frocioneae n. sp. may be distinguished from North and Middle American congeners by the position of the intestinal bifurcation (relatively more posterior in the forebody than in other species) and tegumental spine coverage on the body (spines are absent from the posterior body extremity and on most of the dorsal surface). Comparison of ribosomal DNA (ITS 1 and 2 regions, 5.8S gene, and partial fragment of 28S gene) from the 2 new species and some congeners from the Western Hemisphere provided evidence for the validity of the 2 new species and affirmed a close relationship between H. robisoni n. sp. and Homalometron pallidum. Comparison of ribosomal DNA from newly collected Homalometron spp. and larval stages of an apocreadiid from brackish water hydrobiid snails (cercariae in rediae in Littoridinops palustris and metacercariae in L. palustris and Amnicola limosa) from a tidal river in Mississippi revealed that larval stages represented Homalometron cupuloris. A phylogeny based on Bayesian inference analysis using partial 28S rDNA gene fragments from 14 species of Homalometron (all from the Western Hemisphere) and 1 megaperine and rooted by a second megaperine was conducted and produced a strongly supported phylogram that estimates the interrelationships among species. The estimated phylogeny suggests that ecological factors such as salinity and food web interactions between species of Homalometron, intermediate hosts, and fishes drive coevolutionary forces influencing speciation within Homalometron.

The channel catfish Ictalurus punctatus is a known host for 10 species of Henneguya, but few other myxozoan genera are described from this species. Unicauda is a genus of myxozoan parasites within the family Myxobolidae that consists of 10 valid species from freshwater fish. Herein, we describe a novel species of Unicauda from the intestinal tract of farm-raised channel catfish in Mississippi. Myxospores were consistent with the genus Unicauda but exhibited a unique branching at the terminal end of the caudal process that has not previously been reported. Myxospores measured 90.39 ± 14.97 μm (mean ± SD; range = 70.88–126.02 μm) in total length. The spherical spore body measured 7.31 ± 0.26 μm (6.75–7.84 μm) in length and 7.01 ± 0.63 μm (6.1–8.01 μm) in width. The 2 polar capsules measured 3.45 ± 0.33 μm (3.02–4.03 μm) in length and 2.65 ± 0.32 μm (2.18–3.11 μm) in width. The single caudal process measured 82.98 ± 14.97 μm (63.39–118.63 μm) in length from the base of the spore body to the end of the most terminal projection. Terminal projections measured 26.83 ± 8.8 μm (12.34–42.29 μm) in length and 0.95 ± 0.23 μm (0.52–1.6 μm) in width. The 18S rRNA gene sequence obtained did not match any published sequences. Given the uniqueness of the myxospore morphology, histological presentation, and gene sequence data, we describe this as an unreported species, Unicauda fimbrethilae n. sp.

New records of Macrovalvitrematidae Yamaguti, 1963 are reported from Puerto Rico including Buccamagna archosarga n. gen., n. sp. from the western Atlantic sea bream Archosargus rhomboidalis, Pseudotagia pomadasys n. sp. from the roughneck grunt Pomadasys corvinaeformis, and new locality records for the species Neopterinotrematoides avaginata Suriano, 1975 and Macrovalvitrema argentinensis (Suriano, 1975) n. comb. from the whitemouth croaker Micropogonias furnieri. Buccamagna n. gen. is based on the anterior end and the oral disc being much larger and wider than width of the body proper, by having a pair of rectangular and sclerotized oral suckers, by lacking a pharynx, by having an excessive narrowness in the proximal area of both valves, and by having both valves of the 3 pairs of large clamps armed with several accessory sclerites. Pseudotagia pomadasys n. sp. is distinguished by the absence of serration of lateral sclerites and the relative size of the clamps, having the 2 larger pairs positioned centrally in the haptor and the 2 smaller pairs in the outer positions. Macrovalvitrematoides and Neomacrovalvitrema are synonymized with Macrovalvitrema, which is amended to include Macrovalvitrema micropogoni (Pearse, 1949) n. comb. and Macrovalvitrema argentinensis (Suriano, 1975) n. comb. based on recognition of identical clamp morphologies. In this study the clamp is emphasized as the key taxonomic character for macrovalvitrematid genera. Except for Pseudotagia, the Macrovalvitrematidae have distinctively longer than wide clamps (at least twice as long as wide); with the dorsal valve larger and with a more-complex skeletal structure (g1, g2, i, k, and f sclerites) than ventral valve (c and d sclerites); and 2 clamp morphologies. Characteristics not previously described but present in all macrovalvitrematid monogeneans except Pseudotagia included: (1) asymmetrical plate (plate b) along the ventral valve with a thin fissure on one side, and (2) mirror image arrangement of the pairs of clamps.

This study investigates the systematics of Paracreptotrema Choudhury, Pérez-Ponce de León, Brooks and Daverdin, 2006 using morphological data (stained whole mounts and scanning electron microscopy) and partial sequences of the 28S ribosomal rRNA gene, obtained from freshly collected material. In total, 484 specimens representing 4 species, i.e., Paracreptotrema blancoi (157), Paracreptotrema profundulusi (12), Paracreptotrema rosenthali (8), and Paracreptotrema blancoi sensu Salgado-Maldonado et al. (2011) (307) were collected. Existing museum depositions were also studied. The 28S rRNA gene sequences of these Paracreptotrema spp. were aligned, along with sequences from 22 other allocreadiids and 4 other non-allocreadiid xiphidiatan species. Bayesian inference and maximum likelihood analyses indicated a paraphyletic Paracreptotrema split into 3 clades: 1 comprising P. blancoi and P. rosenthali that was sister to a clade formed by 3 other species of allocreadiids (species of Wallinia, Creptotrematina, and Auriculostoma) typically found in characid fishes, a second clade formed solely by Paracreptotrema heterandriae as the sister taxon of the aforementioned species, and a third by P. profundulusi and specimens erroneously identified as P. blancoi. Two new taxa were erected to reflect these results: Paracreptotrematoides for Paracreptotrema heterandriae, and Pseudoparacreptotrema for Paracreptotrema profundulusi and P. macroacetabulata (the species erroneously identified as P. blancoi from profundulids across Middle America). Closer consideration of the morphology corroborates these findings. The revised systematics also indicated that Paracreptotrema spp. are found in poeciliids, whereas Pseudoparacreptotrema spp. parasitize profundulids. The study demonstrates the value of an integrative taxonomy approach to address the apparently complicated systematics of the allocreadiids.

One of the recalcitrant questions regarding the evolutionary history of clitellate annelids involves the feeding preference of the common ancestor of extant rhynchobdellid (proboscis bearing) and arhynchobdellid (jaw bearing) leeches. Whereas early evidence, based on morphological data, pointed towards independent acquisitions of blood feeding in the 2 orders, molecular-based phylogenetic data suggest that the ancestor of modern leeches was a sanguivore. Here, we use a comparative transcriptomic approach in order to increase our understanding of the diversity of anticoagulation factors for 3 species of the genus Placobdella, for which comparative data have been lacking, and inspect these in light of archetypal anticoagulant data for both arhynchobdellid and other rhynchobdellid species. Notwithstanding the varying levels of host specificity displayed by the 3 different species of Placobdella, transcriptomic profiles with respect to anticoagulation factors were largely similar —this despite the fact that Placobdella kwetlumye only retains a single pair of salivary glands, as opposed to the 2 pairs more common in the genus. Results show that 9 different anticoagulant proteins and an additional 5 putative antihemostasis proteins are expressed in salivary secretions of the 3 species. In particular, an ortholog of the archetypal, single-copy, anticoagulant hirudin (not previously available as comparative data for rhynchobdellids) is present in at least 2 of 3 species examined, corroborating the notion of a single origin of blood feeding in the ancestral leech.

A complete morphological description, supported by molecular data, of Clinostomum detruncatum metacercariae is reported in this study. The metacercariae were found infecting Synbranchus marmoratus from the Paraná River, municipality of Guaíra, Paraná State, Brazil. The taenioidean uterus is the main morphological characteristic of this species. Both neighbor-joining (NJ) and maximum likelihood (ML) trees showed 2 clear separate clusters grouping together the species from the Palearctic region (Clinostomum complanatum, Clinostomum cutaneum, Clinostomum phalacrocoracis, Clinostomum philippinense) and those from the Nearctic/Neotropical regions (Clinostomum marginatum, Clinostomum tataxumui, C. detruncatum, Clinostomum sp. 7). The latter clade is divided into 2 subclades grouping species from North America and Mexico (C. marginatum and C. tataxumui), and those from Brazil (C. detruncatum and Clinostomum sp. 7).

Genotyping of Toxoplasma gondii is traditionally performed using DNA obtained from tachyzoites after isolation by bioassay in mice. In this study, genotyping of T. gondii was performed by multiplex nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) in DNA obtained from the lungs of experimentally infected mice, the hearts of naturally infected free-range chickens, and human blood samples of newborns with congenital toxoplasmosis. The efficiency of Mn-PCR varied according to the marker. We obtained complete genotypes of all of the mice lung samples. In chickens, total or partial genotyping was performed on all of the 15 samples. Two complete genotypes were obtained, including one identified for the first time, and another previously described in different hosts including dogs, cats, and humans. In blood from infants, partial genotypes were obtained in 8 of the 12 samples. Mouse bioassay is the most efficient method to obtain DNA from T. gondii, but direct tissue genotyping enhances the likelihood of obtaining molecular information on T. gondii and is an effective tool as a complement to isolation in mice. In this study, we genotyped Toxoplasma gondii directly from human (blood samples of newborns with congenital toxoplasmosis) and free-range chickens (hearts) by Mn-PCR-RFLP. We present partial and complete genotypes and provide technical and scientific information about T. gondii genotyping methods.

The prevalence and parasitemia of the piroplasm Sauroplasma sp. were evaluated in the Amazon chelonian Podocnemis expansa in Brazil. Samples were collected from 75 chelonians from 3 locations, including a commercial breeding facility, an indigenous subsistence breeding facility, and a wild population. Sauroplasma were found in 72% (54/75) of the chelonians, and the prevalence varied among the sampling sites. No significant correlations were found between the prevalence and the sex and body condition index of the chelonians. The mean parasitemia rate was 44.14/2,000 erythrocytes (2.2%), and no significant correlation was found between the parasitemia and sex and body condition index of the chelonians. These results suggest that the parasite is not pathogenic to P. expansa. No ectoparasites were found in the animals evaluated in the present study; however, due to the aquatic habit of the chelonian, it is likely that the piroplasm is transmitted by leeches and not by ticks, as would be expected for piroplasms.

The scorpion mud turtle (Kinosternon scorpioides) is a small freshwater turtle broadly distributed in South America and commonly consumed in some Brazilian regions. This study aimed to identify the species of helminths that parasitize the digestive tract of K. scorpioides and report infection parameters such as parasite prevalence, mean intensity of the infection, abundance, and the relationship between these nematodes and host body size in this species. We captured 20 adult male K. scorpioides, and 6 animals had nematodes in their gastrointestinal tract. These animals had Serpinema magathi (prevalence = 0.3) and Spiroxys figueiredoi (prevalence = 0.25). There were no correlations between the number of total parasites and carapace length (rs = 0.17, n = 6, P = 0.74) or the length of the gastrointestinal tract (rs = 0.18, n = 6, P = 0.73).