This study examined the relationship between the metazoan parasite community in a focal species, Fundulus heteroclitus, and the species richness of the free-living community. Four distinct salt marsh areas were selected in order to reflect a gradient in free-living species richness and time post-restoration (unrestored, 0, 10, and 20 yr post-restoration). A total of 960 fish was studied, with 30 collected per site in each of 8 consecutive seasons between 2005 and 2007. Species richness and abundance of birds, fish, and benthic macroinvertebrates varied significantly between sites. However, increasing richness in the free-living community was neither positively nor negatively correlated with richness in the parasite infracommunity in F. heteroclitus. These results demonstrate that the relationship between community species richness and parasite richness is not linear. The data suggest that the diversity of parasites may be more correlated with measures of community composition, such as the structure and cohesiveness of the food web, rather than with prevailing measures of biodiversity.

Hairworms infect terrestrial arthropods and are 1 of the most understudied groups of parasites. Recently, life cycles of 2 gordiids (Paragordius varius and Paragordius obamai) have been domesticated in the laboratory. We tested the viability of laboratory reared and post-frozen larval and cyst stages of the North American gordiid, P. varius, frozen at −80 C for 7 mo, and the viability of field collected and post-frozen cysts of the African (P. obamai) and North American (P. varius) gordiid frozen at −20 C for 2 mo. All snails exposed to post-frozen or control P. varius larvae became infected with cysts, and there was no significant difference in prevalence or mean intensity of cysts among control or experimental snail groups. As with larvae, no significant differences were observed in prevalence or mean intensity of emerging worms from crickets infected with post-frozen or control P. obamai or P. varius cysts. All female P. obamai and P. varius worms from control and post-frozen cyst infections laid eggs and larvae hatched from some of these eggs. Survival and cyst formation of P. varius larvae exposed to different combinations of drying and/or freezing temperatures indicated that gordiid larvae have the ability to survive drying and freezing, but survival significantly increases during freezing at lower temperatures. The major contribution of our study is the demonstration that gordiid larval and cyst stages can survive freezing temperatures to infect and develop in the next host.

Populations of several species of gregarine parasites within a single host species, the damselfly Ischnura verticalis, were examined over the course of 1 season at 4 geographic localities separated by a maximum distance of 9.7 km. Gregarines, having a life cycle with both exogenous and endogenous stages, are subject to a wide variety of selective pressures that may drive adaptation. Gregarine species showed some specificity for host life cycle stage, i.e., Steganorhynchus dunwoodyi and Hoplorhynchus acanthatholius were most prevalent in larval hosts while Steganorhynchus dunwoodyi, Actinocephalus carrilynnae, and Nubenocephalus nebraskensis were most prevalent in adult hosts. Species prevalence and abundance differed by geographic locality. Gregarine prevalence was significantly higher in adult female damselflies than in males at 2 localities; sex differences in prevalence were insignificant for larval damselflies at all 4 localities. In larval hosts, gregarine abundance was independent of age (size). The present study, therefore, shows that pond characteristics, host life cycle stage, and adult host sex are the main factors that influence the prevalence and abundance of gregarine populations.

In this study we show that intact DNA can be recovered from both alcohol- and formalin-preserved specimens of the parasitic tapeworm Bertiella studeri for >1 yr after the original fixation and can be successfully amplified and quantified using real-time polymerase chain reaction (RT-PCR). Three different DNA extraction techniques on formalin- and alcohol-fixed material were evaluated to determine which is best suited for obtaining DNA of sufficient yield and purity to be used in downstream applications such as RT-PCR. These techniques included a first organic extraction procedure with an extensive washing step based on a glycine-containing buffer, a second organic extraction procedure that omits the glycine-containing buffer and halves the number of washes in organic solvents, and a third procedure that involves the use of a silica-based DNA binding column from Qiagen^©. The quality of extracted DNA was first examined by agarose gel electrophoresis and ethidium bromide staining and the concentrations were evaluated by OD₂₆₀. We then used an RT-PCR Applied Biosystems TaqMan® Fluorogenic 5′ Nuclease Gene Expression Assay based on the detection of a universal eukaryotic 18S rRNA gene sequence to evaluate the efficiency of the 3 methods. Here we report that, first, the use of a silica column-based DNA extraction technique results in the maximum yield of DNA, on average 55% higher than for the organic extraction methods; second, the use of glycine as a formaldehyde-binding agent in the washing buffer does not necessarily result in a better DNA yield and; third, specimens preserved in ethanol result in significantly higher yields of amplifiable DNA than do specimens preserved in formalin. We, therefore, strongly recommend the use of ethanol as the appropriate fixative if specimens are to be used for molecular work. This is the first report of DNA extraction from preserved specimens of Bertiella studeri.

Aplectana hamatospicula is a nematode that parasitizes the large intestine of anurans, and has been reported from bufonids, eleutherodactylids, hylids, microhylids, and ranids from North and Central America. Aplectana hamatospicula was first described, over 70 yr ago, from Hyla eximia and Bufo peltocephalus from Mexico and Cuba, respectively, and reported from Gastrophryne carolinensis from Florida. Since then there have been no reports of this nematode in North America north of Mexico. The life cycle of A. hamatospicula is not known, and there is limited information on Aplectana spp. from North America. During 2010–2011, we collected 351 anurans of 8 species from 4 locations in Stillwater, Payne County, Oklahoma, U.S.A., and examined them for the presence of Aplectana spp. Of the 8 species of anurans surveyed, A. hamatospicula infected only the Great Plains narrowmouth toad (Gastrophryne olivacea). The prevalence of A. hamatospicula was 85.7% (12/14) with a mean abundance of 28.4 24.0 and a mean intensity of 33.1 22.8 worms per infected toad. We provide new morphological measurements for male and female A. hamatospicula along with new locality records for this nematode in North America.

An examination was made of Giemsa-stained microfilariae in thin blood films from (n = 9) dogs naturally or experimentally infected with Dirofilaria immitis or Dirofilaria repens. Morphological measurements (total length, length of cephalic space, anterior end to nerve ring and last body nucleus, and nucleus-free tail tip) were made on 2–6 microfilariae from each dog with the use of digitally captured images and imaging software. The microfilariae of D. repens were significantly greater (P < 0.001) in all measured dimensions except for the length of the cephalic space, which was significantly shorter (P < 0.001) than that of D. immitis. The cephalic space of D. repens was characterized by being short and routinely being terminated by a distinct pair of nuclei that were separate from the remaining somatic nuclei of the microfilaria. The cephalic space of the smaller microfilaria of D. immitis was longer and did not have the distinct nuclei separated from the somatic column nuclei near the anterior end. The character of the cephalic space seems to be a criterion that could be routinely used for the easy differentiation of these 2 microfilariae in stained blood films.

Trichinellosis has major economic impacts on animal husbandry and food safety, and the control and elimination of trichinellosis is a major objective of veterinary medicine. A gene encoding serine protease of Trichinella spiralis (Ts-Adsp) was identified by immunoscreening an adult T. spiralis cDNA library. In this study, the recombinant Ts-Adsp protein (rTs-Adsp) was cloned and expressed in a prokaryotic expression system and purified by Ni-affinity chromatography. To determine whether the purified rTs-Adsp is a potential vaccine candidate for the control of T. spiralis infection, we immunized BALB/c mice with this protein in combination with an alum adjuvant and subsequently challenged with T. spiralis larvae. The results showed that mice vaccinated with rTs-Adsp exhibited an average reduction in the muscle larvae burden of 46.5% relative to the control group. Immunization with the rTs-Adsp antigen induced both humoral and cellular immune responses, which manifested as elevated specific anti–rTs-Adsp IgG and IgE antibodies and a mixed Th1–Th2 response, as determined by Th1 (IFN-γ and IL-2) and Th2 (IL-4, IL-10, and IL-13) cytokine profiling, with the Th2 predominant. Thus, purified rTs-Adsp is able to limit the invasion of T. spiralis, and this protein could be an effective vaccine candidate for trichinellosis.

Bithynia funiculata is the first intermediate host of the human liver fluke Opisthorchis viverrini in northern Thailand but its identification through morphological analysis is often problematic due to the shortage of gastropod taxonomists. As a consequence, we focused on the development of species-specific primers for use as an identification tool. Our work involved recovery of a 502-base pair (bp) amplicon of unknown function through species-specific primers whose effectiveness was tested by analyzing specimens of B. funiculata from 3 locations in northern Thailand. This primer set did not amplify other species in the Bithyniidae or in other gastropod families. By providing a tool to confirm morphological identifications of B. funiculata, and by enabling the identification of juvenile specimens and those with damaged shells, these primers will improve estimates of the prevalence of parasitic infections in this snail.

Seroprevalences of Toxoplasma gondii antibodies were assessed with the use of a commercial indirect ELISA in 1,501 domestic ruminants in southern Spain. Antibodies against T. gondii were detected in 420 (83.3%) of 504 cattle, 248 (49.3%) of 503 sheep, and 124 (25.1%) of 494 goats. The herd seroprevalence was 100% (72/72), 84.7% (61/72), and 72.2% (52/72) for cattle, sheep, and goats, respectively. Seropositivity was significantly higher in herds with a low density of animals (P < 0.001). Significant differences (P < 0.05) among municipalities were also found. The seroprevalence observed in the present study indicates a widespread exposure to T. gondii in livestock in southern Spain.

During a parasitological survey of naked gobies (Gobiosoma bosc) in Lake Pontchartrain, Louisiana, U.S.A., 2 myxozoan parasites were found infecting the gall bladders of 84 of 90 fish. Parasite plasmodia of each were morphologically distinct; plasmodia bearing Ellipsomyxa-like spores were elongate and fusiform (1,126 by 403 μm), and those bearing Sphaeromyxa-like spores were large and discoid (1,400 μm diameter). Neither parasite conformed to any described species of myxozoan, and both are described here. Sphaeromyxa kenti n. sp. bears boomerang-shaped spores ranging 17.5–19.8 μm long by 3.8–5.2 μm wide. Two unequal sized polar capsules contain ribbon-like polar filaments characteristic of this genus. Ellipsomyxa adlardi n. sp. bears ovoid spores 11.3–14.4 by 7.1–8.8 μm, with 2 rounded polar capsules oriented in opposite directions relative to the longitudinal plane of the spore. Phylogenetically both species cluster with sequenced members of their respective genera. Plasmodia of E. adlardi n. sp. were encountered in 72.2% of gobies examined, and S. kenti n. sp. in 25.6%. Overall, 6.7% of animals were uninfected, and only 4.4% of the 90 fish examined were infected with both species. The observed co-occurrence was lower than expected by chance, strongly suggestive of an interaction between these species.

The need to use complex in vitro cell culture, expensive equipment, and highly-trained technicians that are available only to specialist laboratories has significantly limited studies assessing the potential of pulsed UV light (PUV) to inactivate the waterborne parasite Cryptosporidium parvum in drinking water. This constitutes the first study to report on the use of different non-pathogenic Bacillus endospores as potential surrogate organisms to indicate the PUV inactivation performance of a C. parvum oocyst suspended in water. Findings showed that PUV effectively inactivated approximately 5 log₁₀ CFU/ml Bacillus megaterium and Bacillus pumilus endospores suspended in water at a UV dose of 9.72 μJ/cm² that also inactivated statistically similar levels of C. parvum oocysts (P < 0.05), as determined by combined in vitro HCT-8 cell culture and quantitative PCR. Specifically, this study demonstrated that B. megaterium exhibited greater or similar PUV-inactivation kinetic data compared to that of similarly treated C. parvum over the UV dose range 6.4 to 12.9 μJ/cm². Therefore, the former may be used as an indicator organism for safely investigating the PUV-inactivation performance of this chlorine-resistant, waterborne parasite at the waste-water treatment plant level. Findings presented will impact positively on future water quality studies and on public health.

Neospora caninum is an Apicomplexan parasite that causes repeated abortion and stillbirth in cattle. The aim of this study was to clone the gene encoding the N. caninum orthologue (NcBAG1) of the Toxoplasma gondii bradyzoite-specific protein TgBAG1 and characterize its expression pattern in the parasite. Isolation of the full-length 684-bp gene revealed that it shared 78.3% sequence similarity with TgBAG1. NcBAG1 encodes a predicted protein of 227 amino acids with 80.3% similarity to TgBAG1. A putative signal peptide sequence and an invariant GVL motif characteristic of small heat-shock proteins were identified in the predicted N. caninum amino acid sequence. We expressed the NcBAG1 gene as a recombinant glutathione S-transferase fusion protein (rNcBAG1) in Escherichia coli and used the purified 60 kDa protein to obtain a monoclonal antibody (Mab). rNcBAG1 reacted to Mabs specific for NcBAG1 and TgBAG1. No reaction between the NcBAG1 Mab and N. caninum tachyzoites was observed. Although the predicted molecular mass of NcBAG1 is 25 kDa, Western blot analysis of parasite lysates using the NcBAG1 Mab revealed a cross-reactive protein of approximately 30 kDa. Additionally, immunofluorescence assays using the tachyzoite-specific Mab for NcSAG1 and the bradyzoite-specific Mab for TgBAG1 or NcSAG4 revealed NcBAG1-specific expression in bradyzoites in cultures exposed to sodium nitroprusside, a reagent that increases the frequency of bradyzoites. Interestingly, the NcBAG1 protein was identified in the cytoplasm of the bradyzoite-stage parasites. This preliminary analysis of the NcBAG1 gene will assist investigations into the role of this protein in N. caninum.

Besnoitia besnoiti is an apicomplexan that causes serious economic loss in cattle in many countries and the disease is now spreading in Europe. At least 2 phases of bovine besnoitiosis are recognized clinically. An acute febrile phase characterized by anasarca and necrosis of skin is associated with multiplication of tachyzoites in vascular endothelium; this phase is short-lived and rarely diagnosed. Chronic besnoitiosis characterized by dermal lesions is associated with the presence of macroscopic tissue cysts and is easily diagnosed. Here we report the development of early B. besnoiti tissue cysts in a naturally infected Hugenoot bull from South Africa. Tissue cysts were 10–70 μm in diameter, contained 1–12 bradyzoites, and were most numerous in the dermis, testicles, and pampiniform venous plexus. Amylopectin granules in bradyzoites stained red with periodic acid Schiff (PAS) reaction. Bradyzoites varied in size and in the intensity of PAS reaction (some were PAS-negative), some were plump, and others were slender. With immunohistochemical staining with Besnoitia oryctofelisi and bradyzoite-specific antibodies (BAG-1 made against Toxoplasma gondii bradyzoites), the staining was confined to parasites, and all intracystic organisms were BAG-1 positive. With Gomori's silver stain only bradyzoites were stained very faintly whereas the rest of the tissue cyst was unstained. Ultrastructurally many tissue cysts contained dead bradyzoites and some appeared empty. Unlike bradyzoites from mature cysts, bradyzoites in the present case contained few or no micronemes. These findings are of diagnostic significance. Ultrastructually host cyst cells had features of myofibroblasts, and immunohistochemistry using antibodies against MAC387, lysozyme, vimentin, Von Willebrand factor, and smooth muscle actin confirmed this. Polymerase chain reaction on DNA extracted from lymph node of the bull confirmed B. besnoiti diagnosis. Associated clinical findings, lesions, and histopathology are briefly presented. The bull died of nephrotic syndrome; anasarca in acute besnoitiosis due to protein-losing glomerulopathy is a finding not previously reported in cattle.

A new species of fish leech from Poland, Piscicola burresoni n. sp. (Piscicolidae), is described. The leech was found in 2007 on stones and on pike (Esox lucius L.) and chub (Leuciscus cephalus L.) in the Łyna River, Poland. The leech is small, relatively flattened, with a prominent coarctation between the trachelosome and urosome. The pigmentation is unique to the new species and it has 14 annuli with gonopores separated by 4 annuli. The receptaculum seminis consists of an elliptical aperture located at a distance of 3 annuli behind the female gonopore. The copulatory area on the clitellum is characteristic in the shape of a parallel ellipse. The vector tissue is in the form of a transverse plate below the oviduct outlet. For a complete description of the external morphology, a geometric–mathematical model of the leech body form was used.

Parapharyngodon grenadaensis n. sp. (Nematoda: Pharyngodonidae) from the large intestine of the cane toad, Rhinella marina, is described and illustrated. Parapharyngodon grenadaensis n. sp. is the 48th species assigned to the genus and the 16th species from the Neotropical region. It differs from other species in the genus by possessing 4 pairs of caudal papillae, an echinate anterior cloacal lip, and a blunt spicule of 67–104 μm. This is only the second report of R. marina harboring a species of Parapharyngodon.

The proteocephalidean cestode Harriscolex nathaliae n. sp. (Proteocephalidae: Zygobothriinae) is described from the intestine of the spotted sorubim Pseudoplatystoma corruscans (Spix and Agassiz) (Siluriformes: Pimelodidae) from the Paraná River basin in Argentina. This new species differs from the only species of the genus, Harriscolex kaparari (Woodland, 1935), which is a parasite of Pseudoplatystoma tigrinum (Linnaeus) from the Amazon River in Brazil, by its larger scolex (width of 450–750 μm vs. 305–340 μm), the position of the vagina in relation to the cirrus sac (anterior and posterior vs. only anterior), an asymmetrical vaginal sphincter, the arrangement of vitelline follicles (1 narrow longitudinal band on each side of the proglottid vs. 2 pairs of wide longitudinal bands on dorsal and ventral sides), and a higher number of uterine diverticula (22–45 vs. 16–20). Harriscolex nathaliae is covered with 2 types of microtriches, acicular filitriches and gladiate spinitriches.

Macvicaria magellanica n. sp. (Digenea: Opecoelidae) is the third representative of this genus occurring in fish in the Magellanic sub-region of sub-Antarctica and the first one found in sub-coastal waters in this area (the Beagle Channel). Its main taxonomic features include an elongate body, oral:ventral sucker ratio based on widths of 1.0:1.52–1.98, cirrus sac reaching to level of posterior half of the ventral sucker, testes arranged in tandem, numerous vitelline follicles divided into 2 groups separated by a gap parallel to the ventral sucker, vitelline follicles dorsally not confluent at the uterus and gonads, and egg dimensions of 40–51 × 25–32 μm. The most similar species is Macvicaria antarctica, but it differs from the newly described species in having vitelline follicles dorsally confluent at the level of the uterus and by occurring in fish associated with the Falkland–Patagonian shelf at greater depths (at the North Scotia Ridge, 300–500 m). A key to 9 Antarctic and sub-Antarctic species of Macvicaria is included.

An injured juvenile sharp-shinned hawk, Accipiter striatus Vieillot, 1807 (Aves: Accipitriformes), housed and treated at the College of Veterinary Medicine Teaching Hospital at Kansas State University, Manhattan, Kansas, USA, was found to be passing oocysts of an undescribed species of Caryospora in its feces. Sporulated oocysts of Caryospora petersoni n. sp. were subspherical, with a bilayered wall, and they measured 43.1 × 39.8 μm; micropyle, oocyst residuum, and polar granule were absent. Sporocysts were subspherical to spherical, 23.4 × 23.3 μm; Stieda, substieda, and parastieda bodies were absent, but a spherical sporocyst residuum was present as a compact mass, ∼15.1 μm wide, composed of many homogeneous globules. The new species represents the first caryosporan documented from this species of hawk.

Two mountain beavers, Aplodontia rufa, were collected in Lincoln County, Oregon, and examined for coccidia. Both were infected with 2 new species of Eimeria. Oocysts of Eimeria chitkoae n. sp. were ellipsoidal with a bilayered wall and measured (L × W) 24.5 × 20.2 μm, with a shape index (SI) of 1.2. Both micropyle and oocyst residuum were absent, but a polar granule of several fragments was present. Sporocysts were ovoidal, 12.5 × 7.9 μm, SI was 1.6. Stieda and substieda bodies were present, but a parastieda body was absent; a sporocyst residuum was present, composed of a cluster of moderately coarse granules with many scattered fine granules. Stout sporozoites were 14.7 × 2.9 μm in situ, with spheroidal anterior and posterior refractile bodies. Oocysts of Eimeria lewisi n. sp. were ovoidal, with a smooth single-layered wall, and measured 13.7 × 7.8 μm, SI was 1.7. A micropyle and oocyst residuum were absent, but 1–2 polar granule(s) were present. Sporocysts were 6.6 × 4.2 μm, with SI of 1.6. A Stieda body was present, but substieda and parastieda bodies were absent; a sporocyst residuum was present, composed of a small cluster of several granules. Sporozoites were granular, 8.2 × 1.8 μm in situ, with a posterior refractile body. These are the first coccidians reported from the mountain beaver.

Gravid females of the little-known nematode species Philometra rubra (Leidy, 1856) (Philometridae) are described from specimens from the abdominal cavity of the striped bass, Morone saxatilis (Walbaum), in South Carolina and Canada. The specimens were studied with the use of light and scanning electron microscopy. This species is mainly characterized by the distribution and different sizes of cephalic papillae from the external circle, which is a unique feature compared to other philometrids. Other characteristic features are the shape of the posterior end of body, size and location of caudal projections, and the presence of a well-developed anterior esophageal inflation. The morphology of the male of P. rubra and the life cycle of this nematode are still unknown.

Intromugil alachuaensis n. sp. is described based on specimens collected from the flathead grey mullet (Mugil cephalus) from the Santa Fe River in Florida. The new species is the fourth recognized species in the genus and the second from North America, with the other 2 being confined to South America. Intromugil mugilicolus from Louisiana and Mississippi is redescribed based on the holotype and newly collected material that was not flattened prior to fixation. Two generic features not previously reported are apparent in the new material from I. mugilicolus and I. alachuaensis n. sp.: an armed oral sucker and a series of sacs containing glandular material arranged in symmetrical rows in the hermaphroditic duct. Intromugil alachuaensis differs from I. mugilicolus by having an oral sucker longer than wide, body spines smaller and lanceolate rather than longer and hastate, and smaller vitelline follicles. Intromugil alachuaensis n. sp. differs from Intromugil simonei by having a large elongated pharynx rather than a smaller subspherical one, a proportionately larger and longer oral sucker, and a longer prepharynx (greater than 1 pharyngeal length). Intromugil alachuaensis n. sp. differs from Intromugil annakohnae by having a longer than wide pharynx, a relatively large oral sucker, less extensive vitellarium, and smaller body spines. Comparison of more than 2,400 base-pair-long sequences of nuclear rDNA (partial 18S, complete ITS1, complete 5.8S, complete ITS2, and partial 28S) from I. mugilicolus and I. alachuaensis n. sp. reveals 110 pairwise differences, including gaps, thus supporting our proposal of a new species. These represent the first published sequences from species in this genus.

Oswaldocruzia lamotheargumedoi n. sp., inhabiting the intestine of the cane toad, Rhinella marina (L.), in Laguna de Coyuca, Guerrero, México, is described here. The new species differs from 10 congeners infecting bufonid hosts because it has a type I bursa. In contrast, 7 of these species have type II bursa and 3 more a type III bursa. The species most similar to the species described herein is Oswaldocruzia pipiens Walton, 1929. These 2 species share traits such as body size, bursa type, presence of cervical alae, and dorsal ray morphology. Nevertheless, both species can be distinguished based on the number of synlophe ridges at mid-body (54–56 for O. lamotheargumedoi vs. 45–48 for O. pipiens) and by the presence of a chitinous support in the long, and well developed, cervical alae of O. pipiens. In the new species, these structures are short, poorly developed, and lack chitinous support. Previous records of species of Oswaldocruzia in México include Oswaldocruzia subauricularis (Rudolphi, 1819) Travassos, 1917 in the Neotropical Realm and O. pipiens in the Nearctic.

Here we describe Haemoproteus (Haemoproteus) multivolutinus n. sp. from a tambourine dove (Turtur timpanistria) of Uganda and Haemoproteus (Haemoproteus) paramultipigmentatus n. sp. (Haemosporida, Haemoproteidae) from the Socorro common ground dove (Columbina passerina socorroensis) of Socorro Island, Mexico. These parasites are described based on the morphology of their blood stages and segments of the mitochondrial cytochrome b gene that can be used for molecular identification and diagnosis of these species. Gametocytes of H. multivolutinus possess rod-like pigment granules and are evenly packed with volutin, which masks pigment granules and darkly stains both macro- and microgametocytes in the early stages of their development. Based on these 2 characters, H. multivolutinus can be readily distinguished from other species of hemoproteids parasitizing columbiform (Columbiformes) birds. Haemoproteus paramultipigmentatus resembles Haemoproteus multipigmentatus; it can be distinguished from the latter parasite primarily due to the broadly ovoid shape of its young gametocytes and significantly fewer pigment granules in its fully developed gametocytes. We provide illustrations of blood stages of the new species, and phylogenetic analyses identify DNA lineages closely related to these parasites. Cytochrome b lineages of Haemoproteus multivolutinus and H. paramultipigmentatus cluster with hippoboscid-transmitted lineages of hemoproteids; thus these parasites likely belong to the subgenus Haemoproteus. We emphasize the importance of using cytochrome b sequences in conjunction with thorough microscopic descriptions to facilitate future identification of these and other avian hemosporidian species.

This study documents the prevalences and lineages of hemoparasites in wood stork nestlings from 3 regions of the American continent: southeastern United States (n = 90), northern Brazil (n = 74), and central-western Brazil (n = 125). Identification was based on PCR amplification of a mitochondrial small subunit ribosomal RNA gene. A fragment of the hemoparasite cytochrome B gene in infected individuals was utilized for Bayesian phylogenetic analysis. Four wood stork nestlings were infected by Haemoproteus, 1 from northern Brazil and 3 from the United States, and all shared the same haplotype. Morphological analysis confirmed the infection of the U.S. birds by Haemoproteus. Infection by Plasmodium was found in wood stork nestlings from northern (6) and central-western Brazil (14). Five Plasmodium lineages (MYCAMP1–2, and MYCAMP4–6) were found in the Brazilian central-western region and 3 Plasmodium lineages (MYCAMP2–3, and MYCAMP7) were found in the northern region. The most prevalent haplotype (MYCAMP2) differs from the others by 1 mutation, and the less prevalent haplotypes are derived from MYCAMP2. We did not find Plasmodium or Haemoproteus in nestlings younger than 15 and 30 days old, respectively. This is the first documentation of Plasmodium and Haemoproteus infection in wood storks in Brazilian breeding populations. Potential connectivity among wood stork populations was indirectly supported by the presence of identical Haemoproteus lineages in U.S. and northern Brazilian populations, and by the presence of identical Plasmodium haplotypes in the northern and central-western Brazilian populations.

A new species of monorchiid trematode, Leiomonorchis zhanjiangensis n. sp. (Trematoda: Monorchiidae), is described from the intestine of marine fish Secutor ruconius (Hamilton-Buchanan) (Leiognathidae) from Zhanjiang (21°2′N, 110°3′E), Guangdong Province, China. The new species is distinguished from all known species of the genus, mainly by the configuration of vitelline follicles and the shape of excretory bladder (saccular in the new species vs. tubular in the others). According to Madhavi's opinion, Caballerotrematoides secundus Ahmad, 1981 was transferred from Caballerotrematoides Madhavi, 1977 into Leiomonorchis Mamaev, 1970 as L. secundus (Ahmad, 1981) n. comb.

In Lao People's Democratic Republic (PDR), we detected 7 patients infected with Centrocestus formosanus (1–122 adult specimens) after praziquantel treatment and purgation, together with several other trematode species including Opisthorchis viverrini and Haplorchis taichui. The patients were all men, 23–42 yr-of-age. Three subjects were from Vientiane Municipality and 1 each were from Khammouane, Saravane, Champassak, and Xiengkhouang Province. The patients had frequently eaten raw freshwater fish and were experiencing variable degrees of epigastric pain and indigestion accompanied by occasional diarrhea, although the relationship of these symptoms with C. formosanus infection was unclear. Centrocestus formosanus specimens were ovoid, 0.46 mm (0.41–0.52 mm) long, and 0.18 mm (0.16–0.20 mm) wide (n = 10) and were equipped with 32 circumoral spines on the oral sucker. The uterine eggs were 33.2 μm long (31.8–34.9 μm) and 18.5 μm wide (17.4–19.8 μm) (n = 20). Analysis of the nucleotide sequence of the 18S ribosomal RNA gene of our specimens (Laotian isolate) revealed 100% homology with that of an isolate from the United States reported in GenBank. Several species of freshwater fish collected from Xiengkhouang Province revealed a 17.0% prevalence (9 of 53 fish examined) for C. formosanus metacercariae. The results suggest that human C. formosanus infections have been masked by other trematode infections.

The seroprevalence of Toxocara canis antibodies in children aged from 1 to 12 yr old was evaluated in Pelotas City, Rio Grande do Sul, Brazil. Human toxocariasis or visceral larva migrans (VLM) was diagnosed with the use of an ELISA based on the T. canis excretory–secretory (TES) antigens; Western blotting was used to confirm the ELISA-positive results. From 427 samples, 50.6% were positive for the presence of anti-TES antibodies. A confirmatory test (Western blot) was carried out on a sample of the ELISA-positive sera (n = 70), and all were positive. The Western blots had specific banding pattern characteristics, where the 30-kDa fraction demonstrated the highest reactivity. This fraction could be important for the specific diagnosis of toxocariasis.

Seroprevalence of Toxoplasma gondii infection in goats in Michoacán, Mexico, is largely unknown. Antibodies to T. gondii were determined in 341 dairy goats in Michoacán, Mexico, using the modified agglutination test. Goats were raised in 9 farms in 6 municipalities. Overall, antibodies to Toxoplasma were found in 52 (15.2%) of 341 goats, with titers of 1:25 in 16, 1:50 in 9, 1:100 in 4, 1:200 in 4, 1:400 in 4, 1:800 in 9, 1:1,600 in 3, and 1:3,200 or higher in 3. Seropositive goats were found in all 9 farms sampled, and seroprevalence varied significantly among farms (1.9–90%). Seroprevalence of T. gondii varied with age, municipality, altitude, and climate but not with breed. Increased seroprevalence was found in goats aged 13–24 and 49–86 mo old (25% and 22.9%, respectively). Goats raised in farms in a municipality with semi-warm humid climate at 1,700 m of altitude had the highest seroprevalence (62.1%). This is the first report of T. gondii infection in goats in Michoacán State, Mexico, and of an association of seropositivity to T. gondii and semi-warm humid climate. Results indicate that infected goats are likely an important source of infection with T. gondii in humans in Michoacán State.

We used a Biomphalaria glabrata snail model for our studies and investigated the suitability of B. glabrata neonates, reared on a Nostoc sp. diet, for infection with Echinostoma caproni miracidia. We found that neonatal snails could become infected with E. caproni miracidia with 31 ± 11% standard error (SE) of our exposed snails containing rediae infections at 4 wk post-exposure (PE). However, the survival of exposed neonates was significantly (P < 0.05, Student's t-test) less than that of the unexposed controls at 1, 2, 3, and 4 wk PE.

Seroprevalence of Toxoplasma gondii infection in fattening pigs in Heilongjiang Province, northeast China, was investigated between July 2011 and June 2012. In total, 1,014 pig serum samples were collected from 10 administrative regions and assayed for T. gondii antibodies by indirect hemagglutination (IHA) test using a commercially available kit. The average T. gondii antibody-positive prevalence was 4.6% (47/1,014), which is relatively lower than that in other provinces in China. Seroprevalence in fattening pigs raised on small farms (6.3%) was significantly higher than that on large farms (3.8%) (P < 0.05), and the seroprevalence ranged from 2.4% (Shuangyashan) to 6.8% (Qitaihe) between different geographical regions. Given that pork is not examined for T. gondii infection in China by law, these findings have public health implications and provide useful baseline information for the control of pig toxoplasmosis in this unique region of China.

Lice infestations are commonly seen in buffaloes, causing damage directly to the animal, i.e., itching, skin lesions, and anemia. In addition, these insects can also be vectors for infectious diseases. The present study describes an outbreak of lice in buffaloes as well as evidence for Haematopinus tuberculatus acting as a vector of anaplasmosis. Lice and blood were collected from 4 young buffaloes (2- to 4-mo-old) and a molecular analysis for the presence of Anaplasma marginale was conducted. DNA of A. marginale was detected in the blood of all 4 animals. Twelve lice were collected and separated in 4 groups, with 3 insects each, to comprise a pool of samples. After DNA extraction and molecular analysis, a positive PCR for A. marginale was found in all pooled samples. These results identify sucking lice as potential vectors of anaplasmosis. However, additional studies are necessary to fully evaluate the vector potential of H. tuberculatus for A. maginale transmission.

The geographic structuring of parasite communities across the range of a single host species can illuminate patterns of host-population connectivity. To determine the location of parasite transmission in a Neotropical migrant bird species, we sampled adult and hatch-year (HY) birds across the breeding and wintering range of the Swainson's thrush (SWTH), an abundant passerine with a migratory divide. We examined the phylogenetic relationships among cytochrome b lineages of the avian blood parasite genera Haemoproteus, Plasmodium, and Leucocytozoon and determined the transmission location of unique lineages. We found that Haemoproteus and Plasmodium lineages are transmitted on California breeding grounds, whereas Leucocytozoon transmission occurs on Alaskan breeding grounds. The presence of hemosporidians on wintering grounds and shared lineages between the SWTH and resident species suggests that transmission of some of these lineages occurs on both breeding and wintering grounds. We emphasize that the sampling of HY birds and local resident heterospecifics will supplement vector studies to determine the key players in hemosporidian host switching and range-expansion events.

The aim of this study was to evaluate Cryptosporidium spp. contamination of sea and tap water samples from Sinop and Ordu Provinces, Black Sea, Turkey. The samples (10 L) were collected in spring, summer, autumn, and winter in 2011. A total of 128 water samples was analyzed using an immunofluorescence test (IFT), as well as loop-mediated isothermal amplification (LAMP) and nested polymerase chain reaction (PCR). Cryptosporidium spp. oocysts were detected by IFT in 43 of the 70 samples (61.4%; 1–40 oocysts per 0.5 L) and 35 of the 58 samples (60.3%; 1–23 oocysts per 0.5 L) in the sea water samples from Ordu and Sinop, respectively. The highest number of oocysts by IFT were detected in spring and winter in Ordu and Sinop, respectively. The results of the S-adenosylmethionine synthetase (SAM) gene LAMP assays were 65.5% positive for Cryptosporidium parvum, Cryptosporidium hominis, and Cryptosporidium meleagridis in all examined samples, while the SSUrRNA gene nested PCR assay was 31.0% positive. Six C. parvum nested PCR products from all positive samples were successfully sequenced.

Species of Haematoloechus and Rhabdias both are ubiquitous lung parasites of frogs, yet surprisingly little research has been conducted on the interactions between these worms. In a small Nebraska stream, 256 Rhabdias joaquinensis and 225 Haematoloechus complexus were found in the lungs of Lithobates (=Rana) blairi. Thirty-six of the 44 (82%) frogs were co-infected with H. complexus and R. joaquinensis whereas 5 (11%) frogs were infected with only 1 species and 3 (7%) frogs were uninfected. Tests for association between H. complexus and R. joaquinensis found a significant positive relationship between the 2 parasites. Significant positive relationships in intensity of infection were found when both lungs were combined but not when lungs were considered individually. Tests for association between H. complexus and R. joaquinensis found no relationship between the worms in the anterior and posterior portions of the lungs. These data suggest that H. complexus and R. joaquinensis do not competitively exclude each other from the lungs of L. blairi in southeastern Nebraska.

Understanding the spread of Toxoplasma gondii in waterfowl is of interest for elucidating the potential involvement of these birds in maintaining the parasitic life cycle because birds are exposed to these parasites. Sera from 103 adult, free-range game birds, representing 13 different species living in the Italian wetlands, were examined using modified agglutination tests for antibodies specific to T. gondii. In seropositive birds, the brain and heart were homogenized and DNA was extracted to perform nested polymerase chain reaction (nPCR) and genotyping. Out of the 103 serum samples, 9 (8.7%) were scored as positive with titers ranging from 1/20 to 1/160. The seropositive animals corresponded to 4 of the species: Anas platyrhynchos (mallard; 2/17), Anas clypeata (northern shoveller; 2/11), Anas crecca (common teal; 3/41), and Gallinago gallinago (common snipe; 2/8). Parasite DNA was detected in 3 out of 9 brain samples, while the PCR results from the heart specimens were negative in all of the birds. The occurrence of non-clonal types of T. gondii was suggested in all cases. In conclusion, we describe the first documented detection of DNA of T. gondii in tissues from the northern shoveller and common teal, thereby extending the range of intermediate hosts for this parasite.

The amphibian genus Leptodactylus includes around 50 species, of which only 2 are distributed in Mexico; the helminth fauna of these 2 species is poorly known. As part of a research program on amphibian parasites in Mexico from 1997 to 2005, 281 sabinal frogs Leptodactylus melanonotus from 42 localities in 11 Mexican states were examined from a helminthological perspective. A total of 20 taxa of helminths—7 digeneans (5 adults, 2 larvae) and 13 nematodes (8 adults, 5 larvae)—was found to infect this amphibian host species. These data represent 105 new locality records, and 11 taxa are recorded in L. melanonotus for the first time. Infracommunity analyses of the sabinal frogs from Tres Palos indicated that these hosts are depauperate. The helminth community is dominated by specialist species, with Cosmocerca podicipinus the most common in almost 50% of the infracommunities. Percutaneous infection and predator-prey interactions were the 2 most common infection routes by helminths in frogs from Tres Palos, with 79% of the parasites recruited via skin penetration. Finally, our results show that the helminth fauna parasitizing L. melanonotus throughout Mexico has low similarity with the helminth fauna of leptodactylids studied comprehensively in South America, with only 2 digeneans and 3 nematodes being shared by hosts from both regions. As a result of our survey, the number of helminth species parasitizing L. melanonotus increased to 34. Considering its native distribution range, this number is now 36 with the inclusion of the nematodes Oswaldocruzia costaricensis and Cruzia empera in Costa Rica.

Tapeworm eggs from the genus Taenia sp. were identified during the study of mummy remains dated to 2,286 ± 28 BP from the Chehrabad salt mine in northwestern Iran. The presence of tapeworm in this salt mine provides paleopathological information. Moreover, it brings new information on ancient diet, indicating the consumption of raw or undercooked meat. Cultural aspects as well as archaeozoological data are discussed in order to try to detail meat consumption. Paleoparasitological data are rare in the Middle East, and this case study presents the first recovery of parasites in ancient Iran. It constitutes the earliest evidence of ancient intestinal parasites in this country and contributes to the knowledge of gastrointestinal pathogens in the Near East.

Protozoans of the family Opalinidae are intestinal commensals in amphibians. To test the hypothesis that these organisms are susceptible to the antiprotozoal antibiotic metronidazole, we randomly assigned 60 juvenile Woodhouse's toads (Bufo woodhousii) to receive a single oral dose of metronidazole or water. In pilot trials, the prevalence of opalinids in untreated members of this population was over 70%. One-third of the study population was dissected at each of 3 time points: 18 hr, 1 wk, and 2 wk post-treatment. An examiner blinded to the toad's treatment history determined the presence or absence of opalinids using a dissecting microscope. Opalinids were found in 3/10 toads in the treatment group and 9/10 in the control group after 18 hr (P < 0.02), in none of the treatment group and 8/10 in the control group after 1 wk (P < 0.001), and in none of the treatment group and 10/10 in the control group after 2 wk (P < 0.0001). These results suggest that a single-dose of metronidazole quickly and reliably clears opalinids from juvenile Woodhouse's toads with no evidence of short-term recurrence. The treatment was well tolerated, with no apparent morbidity and no mortality in either group. Future exploration of opalinid-related host fitness consequences may be facilitated by this simple method of developing a protozoan-free host population.

We report on the intestinal helminth fauna of 15 bottlenose dolphins Tursiops truncatus and 6 short-beaked common dolphins Delphinus delphis from the western Mediterranean. Eight helminth species were found in bottlenose dolphin, i.e., the digeneans Synthesium tursionis, Brachycladium atlanticum, and Pholeter gastrophilus, the nematode Anisakis sp., and the cestodes Tetrabothrius forsteri, Diphyllobothrium sp., Strobilocephalus triangularis, and tetraphyllidean plerocercoids. Brachycladium atlanticum, S. triangularis, and tetraphyllidean plerocercoids are new host records. No T. forsteri had previously been reported in Mediterranean bottlenose dolphins. Three species of helminths were recorded in the common dolphin, i.e., the digenean Synthesium delamurei (which was a new host record), and the cestodes T. forsteri and tetraphyllidean plerocercoids. The intestinal helminth communities of bottlenose and common dolphins are depauperate, similar to that of other cetacean species, but those from bottlenose dolphins harbored a higher number of helminth species. This study supports the notion that oceanic cetaceans, such as common dolphins, have a comparatively poorer helminth fauna than that of neritic species, such as bottlenose dolphins, because the likelihood of parasite recruitment is decreased.

Toxoplasma gondii is an important protozoan parasite that can infect virtually all warm-blooded animals, including humans. Oocysts in the environment are an important infection source. Microtus fortis can be used as a sentinel reflecting the distribution of T. gondii oocysts in the environment indirectly. In the present study, we developed a seminested polymerase chain reaction (PCR) targeting T. gondii 529–base-pair repeat sequence, with high sensitivity and specificity. Two hundred seventy-two free-living M. fortis were collected from Jilin Province, Northeastern China, and the lung samples were examined for T. gondii DNA by seminested PCR. Toxoplasma gondii DNA was found in 137 (50.4%) of these rodents. The high prevalence of T. gondii infection in free-living M. fortis suggests a wide distribution of T. gondii oocysts and a high infection pressure for humans and other animals.

In 2004, an article in the Journal of the New York Entomological Society claimed that individuals with delusory parasitosis actually suffer from collembola infestations. The article has been critiqued for poor methodology and results that contradict all knowledge about collembolans. Less easily accounted for has been a figure in the article purporting to show a collembolan in a skin scraping. The image appears to have been altered using photo manipulation software to an unacceptable degree, and this paper demonstrates that to be the case. The altered figure represents creation of nonexistent data, a form of scientific misconduct. Whether the deception is deliberate or a product of an otherwise well-meaning author ignorant of the limits of acceptable image manipulation is unknown, but the result is peer-reviewed support for a conclusion that complicates patient treatment. In the current era of computers, even regional entomology journals must have detailed standards for what kind of images and image manipulations are acceptable for publication.