Myxozoans are a clade of highly derived cnidarians. The phylogenetic identity of these extremely simplified parasites of aquatic vertebrates and invertebrates had long been uncertain, with all early classifications designating Myxozoa as protists. Though suggestions were frequently made that the infective spores of these parasites are multicellular and possibly of cnidarian origin, it would take a phylogenetic analysis of ultrastructural developmental characters in combination with rRNA gene sequences to verify the Myxozoa as secondarily reduced cnidarians, sister to the polypoidozoan parasite Polypodium hydriforme. While a series of subsequent molecular studies suggested hypotheses of Myxozoa as basal bilaterians, triploblasts, or even nematodes, phylogenomic analyses with improved taxon sampling corroborated the landmark paper that verified the cnidarian nature of this group. This review of the body of phylogenetic work on Myxozoa aims to clarify historical progress and current knowledge, as well as to emphasize the opportune position that myxozoan biologists now are in, to address fundamental questions of cell biology of these parasites as well as the evolution of animal life.

Given their ubiquitous nature, it is surprising that more oligochaete annelid worms (Annelida: Clitellata) have not adopted an endoparasitic lifestyle. Exceptions, however, are the understudied members of the genus Dero (Allodero) that parasitize the ureters of tree frogs and toads. This study experimentally explores the life cycle and host specificity of Allodero hylae, the worm's use of chemical cues in host searching, and its seasonal prevalence and abundance over a year-long collection period on the Florida Southern College campus. A total of 2,005 A. hylae was collected from the ureter, urinary bladder, or expressed urine of wild Osteopilus septentrionalis; a significant positive correlation was found between host snout–vent length and parasite intensity for female but not male hosts. Monthly prevalence of A. hylae reached a peak of 58% in April, but never dropped below 20% in any month; mean abundance peaked March–May, whereas few worms were recovered in December and January. Confirming a parasitic lifestyle, wild-collected hosts with intense infections, typically >40 worms, showed obvious dilatation of the ureter wall, and some young-of-the-year O. septentrionalis exposed to A. hylae in the laboratory were killed by the apparent rupture of the host's ureter. The worm has a direct life cycle: worms expelled in the host's urine are capable of locating and re-infecting other hosts within aquatic microhabitats such as bromeliad tanks, and worms can survive for weeks in a free-living environment, even undergoing a morphological change. Further, chemotaxis assays found a positive response to a tree frog attractant for worms recently removed from hosts. Overall, this study provides the first multifaceted investigation on the life history and ecology of any Allodero spp., which offers new insights into an understudied endoparasitic oligochaete.

Parasite assemblages acquired through trophic interactions in fish hosts are increasingly cited as a means to determine pollution effects on water quality and food web structure. We examined gastrointestinal parasite community changes above and below coal mine input from 597 individuals representing 3 species of sunfish: green sunfish (Lepomis cyanellus), bluegill (L. macrochirus), and longear sunfish (L. megalotis). Hosts were collected from 6 sites in or near the south fork of the Saline River Basin in southern Illinois in the spring and fall of 2006. Three sites received no known effluent from coal mines. An additional 3 sites received effluent termed acid mine drainage (AMD). We recovered 1,064 parasites from 12 genera. The parasite community in sunfish collected downstream nearest to the source of AMD was significantly different from 3 upstream sites. In addition, 2 sites farther downstream receiving AMD were different from 2 of 3 reference sites. However, there was also considerable variability in parasite assemblages between sites grouped as above or below coal mine effluent. Several parasite species responded to changes in water quality. Spinitectus sp. (Nematoda), which uses sensitive mayfly hosts to complete its life cycle, was less abundant at sites downstream of coal mine effluent in both green sunfish and bluegill. In contrast, 2 acanthocephalans (Neoechinorhynchus sp. and Eocollis arcanus) and a nematode (Spiroxys sp.) were found in green sunfish more frequently in areas downstream of AMD. This study further suggests metazoan parasites may be useful as indicators of water quality; however, variability among similar sites may limit their application. In addition, strong assemblage differences were found among the 3 sunfish species, suggesting variable habitat usage and potential resource partitioning among congeneric fish hosts in streams.

Biological invasions can bring both the invader and native taxa into contact with novel parasites. As cane toads (Rhinella marina) have spread through Australia, they have encountered lungworms (Rhabdias hylae) that occur in native frogs. Field surveys suggest that these lungworms have not host-switched to toads. In our laboratory studies, R. hylae infected cane toads as readily as it infected native frogs, but failed to reach the lungs of the novel host (i.e., were killed by the toads' immune response). Plausibly, then, R. hylae might reduce the viability both of their native hosts (frogs, that can exhibit high parasite burdens) and cane toads (that must deal with infective larvae traveling through the host body). Our laboratory trials suggest, however, that the impacts of the parasite on infected anuran hosts (both frogs and toads) were minimal, with no significant decrements to host survival, activity, growth, or locomotor performance. Ironically, the lack of impact of the parasite on its native hosts appears to be an outcome of co-evolution (frogs tolerate the lungworm), whereas the lack of impact on the novel host is due to a lack of co-evolution (toads can recognize and eliminate the lungworm).

Echinococcus multilocularis (EM) is a pathogenic and potentially fatal cestode causing human alveolar echinococcosis (AE). A meta-analysis was conducted using a generalized estimation equation approach (GEE) to assess the effect of taxonomic, environmental, and diagnostic variables on EM prevalence in different hosts. Red foxes (Vulpes vulpes) had significantly higher prevalence of EM than domestic dogs (Canis lupus familiaris), with the diagnostic method playing an important factor in assessing prevalence. For intermediate hosts genera was significantly associated with EM prevalence, although there was some indication of publication bias in this dataset. This study also highlights the possible importance of temperature and precipitation to EM transmission. This implies the possibility of a changing climate affecting the future distribution of the parasite.

The chewing louse fauna of pigeons and doves in Japan is reviewed based on published records and new collections. An updated checklist of the chewing lice of Japanese pigeons and doves is provided, and 3 new species are described: Columbicola asukae n. sp. and Coloceras nakamurai n. sp., both from Columba janthina Temminck, 1830 (Japanese wood pigeon), and Columbicola lemoinei n. sp. from Treron formosae permagnus Stejneger, 1887, and Treron formosae medioximus (Bangs, 1901) (whistling green-pigeons). This checklist includes data on the first records of Coloceras chinense (Kellogg and Chapman, 1902), Coloceras piriformis (Tendeiro, 1969), and Columbicola guimaraesi Tendeiro, 1965, in Japan. New host records of Hohorstiella sp. from Columba janthina and Treron formosae permagnus, and Coloceras sp. from Treron sieboldii sieboldii (Temminck, 1835) (white-bellied green-pigeon) are provided.

Three new gamasid mite species belonging to the genus Macronyssus Kolenati, 1858 (Acari: Macronyssidae), namely, Macronyssus sibiricus n. sp., Macronyssus stanyukovichi n. sp., and Macronyssus tigirecus n. sp., are described (females only; males, protonymphs, and larvae remain unknown). All species are known from Western Siberia and belong to the Siberian–Far Eastern bat ectoparasite fauna complex. The parasite hosts are the eastern water bat Myotis petax Hollister, 1912, and Hilgendorf's tube-nosed bat Murina hilgendorfi Peters, 1880 (Chiroptera: Vespertilionidae). An identification key for females of the genus Macronyssus Kolenati, 1858, in the boreal Palearctic region is presented.

Within Oligochaeta, Chaetogaster limnaei is unusual in exhibiting a parasitic relationship with freshwater pulmonate snails. Taxonomic confusion has been caused by differences in what have been considered 2 subspecies of this worm: Chaetogaster limnaei limnaei is an ectosymbiont and is present inside the mantle cavity of the snail, whereas Chaetogaster limnaei vaghini is parasitic and lives in the kidney of the snail. This study explored the distribution of these annelids in central New York and used mitochondrial DNA sequence data from the COI locus to examine the relationship, evolution, and species status of the ectosymbiotic and parasitic forms of C. limnaei. Snails (Physa gyrina) were collected from 6 streams and lakes in central New York, with additional specimens collected from a lake in Massachusetts for comparison. One hundred and forty snails were examined, and at least 1 form of Chaetogaster was present in 88 specimens, a prevalence of 62.9%. COI sequence data from New York and Massachusetts did not reveal separate ectosymbiotic and parasitic lineages. Instead, all parasitic forms were part of a mixed clade that included both ectosymbiotic and parasitic forms. This mixed clade was nested within clades of ectosymbiotic forms only, suggesting that a plastic lineage of C. limnaei, able to be both ectosymbionts and parasites, evolved from ectoparasitic ancestors.

The relative importance of environmental factors and host factors in explaining variation in prevalence and intensity of flea parasitism in small mammal communities is poorly established. We examined these relationships in an East African savanna landscape, considering multiple host levels: across individuals within a local population, across populations within species, and across species within a landscape. We sampled fleas from 2,672 small mammals of 27 species. This included a total of 8,283 fleas, with 5 genera and 12 species identified. Across individual hosts within a site, both rodent body mass and season affected total intensity of flea infestation, although the explanatory power of these factors was generally modest (<10%). Across host populations in the landscape, we found consistently positive effects of host density and negative effects of vegetation cover on the intensity of flea infestation. Other factors explored (host diversity, annual rainfall, anthropogenic disturbance, and soil properties) tended to have lower and less consistent explanatory power. Across host species in the landscape, we found that host body mass was strongly positively correlated with both prevalence and intensity of flea parasitism, while average robustness of a host species to disturbance was not correlated with flea parasitism. Cumulatively, these results provide insight into the intricate roles of both host and environmental factors in explaining complex patterns of flea parasitism across landscape mosaics.

Trichuris suis is a common parasitic helminth of pigs. As with many other parasites, T. suis ensures its own survival by evading host immune responses, but little is known about how this is achieved. MicroRNAs (miRNA) have been shown to be involved in various immunological processes by post-transcriptional regulation of specific genes, and the potential of using these molecules as biomarkers of disease is currently being examined. It has recently been shown that parasites may secrete extracellular structures such as exosomes and microvesicles, containing proteins and miRNA. The fusion of these structures with host cells has been demonstrated, and a role of exosome-derived miRNA in host gene regulation has been suggested. In the present study, we show that exosome- and microvesicular-like structures are secreted by T. suis L1 larvae and also demonstrate the presence of miRNA-sized RNA inside these structures. A potential role of these molecules in host-parasite interactions is suggested. In addition, an electron-dense layer covering the surface of the larvae was observed, which may play a function in host immune evasion.

An analysis of gastrointestinal parasites of Ecuadorian mantled howler monkeys, Alouatta palliata aequatorialis, was conducted based on examination of fecal smears, flotations, and sedimentations. At least 1 type of parasite was detected in 97% of the 96 fecal samples screened across 19 howler monkey groups using these techniques. Samples averaged 3.6 parasite species per individual (±1.4 SD). Parasites included species representing genera of 2 apicomplexans: Cyclospora sp. (18% of individual samples) and Isospora sp. (3%); 6 other protozoa: Balantidium sp. (9%), Blastocystis sp. (60%), Chilomastix sp. (4%), Dientamoeba sp. (3%), Entamoeba species (56%), Iodamoeba sp. (5%); 4 nematodes: Enterobius sp. (3%), Capillaria sp. (78%), Strongyloides spp. (88%) which included 2 morphotypes, Trypanoxyuris sp. (12%); and the platyhelminth Controrchis sp. (15%). A statistically significant positive correlation was found between group size and each of 3 different estimators of parasite species richness adjusted for sampling effort (ICE: r² = 0.24, P = 0.05; Chao2: r² = 0.25, P = 0.05, and Jackknife: r² = 0.31, P = 0.03). Two significant associations between co-infecting parasites were identified. Based on the prevalence data, individuals infected with Balantidium sp. were more likely to also be infected with Isospora sp. (χ² = 6.02, P = 0.01), while individuals harboring Chilomastix sp. were less likely to have Capillaria sp. present (χ² = 4.03, P = 0.04).

The epidermal mucus covering the surface of a snail represents an important barrier to trematode larvae attempting to penetrate the snail and may play a role in mediating snail-trematode compatibility. In this study, Facioloides magna miracidia were exposed to mucus harvested from a compatible snail host, Lymnaea elodes (palustris), and from an incompatible snail, Helisoma trivolvis. In vitro treatment of freshly hatched miracidia with snail-derived mucus exerted dramatically different effects on larvae depending on snail species. At the lowest dilution of mucus tested (1:3) mean damage rates (tegumental damage and/or larval lysis and death) were as high as 100% for miracidia exposed to H. trivolvis mucus, while none of F. magna miracidia were damaged in L. elodes mucus. A dilution series for each snail species, and treatments with heat and proteinase K were performed to characterize the component(s) of mucus inducing the observed morphological changes. The damaging effects of H. trivolvis mucus were concentration dependent and completely abrogated by heat (65 C, 30 min) and proteinase treatment, strongly implicating a heat-labile protein(s) in mucus as the active cytotoxic agent(s). In contrast to our prediction that miracidial contact with mucus of compatible L. elodes would trigger larval transformation, mucus from either snail species tested exhibited little to no activity. Overall these data demonstrate the presence of a potent cytotoxic protein-like factor in the mucus of F. magna–incompatible H. trivolvis, and its absence in the mucus of the compatible snail, L. elodes. This finding supports the notion that the epidermal mucus layer may be serving as an important determinant of larval trematode-snail compatibility.

Although lacking an NADPH→NAD transhydrogenase system, the essentially energetically anaerobic mitochondria of the adult intestinal nematode Ascaris suum display an inner membrane-associated NADH→NAD transhydrogenation reaction. This reaction is considered to be reflective of a mechanism(s) that acts in catalyzing a transmembrane translocation of reducing equivalents from NADH in the intermembrane space to matrix NAD , thereby forming matrix NADH that would serve in electron transport. Ascarid mitochondrial lipoamide dehydrogenase rather than an NADH→NAD transhydrogenase system has been viewed as the predominant source of inner membrane-associated NADH→NAD transhydrogenation activity. However, the present study made apparent yet another source of mitochondrial, inner membrane–associated NADH→NAD activity in A. suum, viz., NADH dehydrogenase. This was made evident via comparisons of the A. suum mitochondrial NADH→NAD transhydrogenation, NADH dehydrogenase, and lipoamide dehydrogenase activities in terms of pH effects, thermal labilities, the involvement of NADH dehydrogenase in the activities of mitochondrial, membrane-associated rotenone-insensitive and rotenone-sensitive NADH-dependent cytochrome c reductases, and mitochondrial membrane versus mitochondrial soluble localizations. Studies of the responses of the NADH→NAD transhydrogenation, rotenone-insensitive and rotenone-sensitive cytochrome c reductases, and lipoamide dehydrogenase activities to inhibition by copper and cadmium lent additional support to the catalysis of an NADH→NAD transhydrogenation activity by NADH dehydrogenase. Collectively, the data presented are consistent with an additional physiological catalysis of an NADH→NAD transhydrogenation in A. suum mitochondria by an inner membrane NADH dehydrogenase component of the rotenone-sensitive cytochrome c reductase system, i.e., the NADH dehydrogenase component of the electron transport system. Comparisons of the A. suum data with those from other essentially anaerobic helminth parasites as well as free-living eukaryotic mitochondrial systems are noted.

The effects of Oxyspirura petrowi infections in northern bobwhites (Colinus virginianus) are not well understood. While studies have reported O. petrowi infections, none has histopathologically examined the eye surface and intraorbital glands to assess cellular-level impacts associated with infection. This study is the first to document the histopathology associated with O. petrowi infections. Oxyspirura petrowi occurred on the eye surface as well as in the conjunctiva, lacrimal ducts, lacrimal glands, and Harderian glands. Histopathology showed infections of O. petrowi caused cellular damage to these tissues, scarring and interstitial keratitis of the cornea, and acinar atrophy of the Harderian gland.

Our main aims were to investigate hypoxia inducible factor-1α (HIF-1α) expression in the surrounding invasion range of hepatic alveolar echinococcosis (HAE) lesions and determine the pathological basis of angiogenesis. In total, 23 Wistar rats with hepatic echinococcus multilocularis infection were killed and their livers, which contained 27 HAE lesions, obtained. Specimen segments were generated from 119 paraffin blocks. Comparative analysis of the tissue samples containing HAE nodules and hepatic parenchyma of the surrounding region was performed with the immunohistochemical SP method in this animal experiment. Expression patterns of HIF-1α in the surrounding invasion range and the hepatic parenchyma were compared. The HIF-1α positive expression rate was 97.5% (116/119 samples). Expression of HIF-1α in the actively multiplying infiltrative region of the HAE lesions was significantly higher than that in hepatic parenchyma (P < 0.05). Overexpression of HIF-1α in the actively multiplying infiltrative region of HAE lesions in rats is closely related to angiogenesis and microvasculature. The sensitivity of HIF-1α facilitates its application as a representative maker of HAE. Our data indicate that the invasion range of HAE lesions is based on extrusion and compression, and induces anoxia and ischemia in hepatic tissue. Thus, HIF-1α provides a valuable index for evaluating HAE activity, and induces anoxia and ischemia in hepatic tissue.

Parapharyngodon guerreroensis n. sp. (Nematoda: Pharyngodonidae) from the large intestine of Lepidophyma smithii from Mexico is described and illustrated. A list of nominal species and a key to species from the Panamanian region are provided. Parapharyngodon guerreroensis n. sp. is the 57th species assigned to the genus and the 10th from the Panamanian region. It differs from other species in the genus in that males possess 3 pairs of caudal papillae, an anterior cloacal lip supporting 4 digitiform processes, and a blunt spicule 67–104 μm in length, while females possess long flexible caudal appendages.

Heligmosomoides americanus is shown by molecular phylogenetic analysis of 3 nuclear (28S, ITS1, and ITS2) and 2 mitochondrial (cytochrome oxidase 1 and cytochrome b) loci to be a distinct species of heligmosomid nematode with a long-independent evolutionary history, and not a subspecies of Heligmosomoides polygyrus. Rather than being a recent arrival in North America, the species probably originated as a Beringian immigrant with the host vole Phenacomys, approximately 2 million years ago (MYA).

Massive numbers of sarcocysts of a previously undescribed species of Sarcocystis were observed in the skeletal muscles throughout the body of an adult, female South American rattlesnake (Crotalus durissus terrificus). Examination of tissue sections by light microscopy demonstrated that sarcocysts were present in 20 to 40% of muscle fibers from 5 sampled locations. Sarcocysts were not present in cardiac muscle, smooth muscle, or other organs. Sarcocysts were 0.05–0.15 mm wide, had variable length depending on the viewed orientation and size of the muscle fiber, and had a sarcocyst wall less than 1-μm thick. Sarcocysts were subdivided by septa and had central degeneration in older sarcocysts. Host induced secondary encapsulation or an inflammatory response was not present. By transmission electron microscopy (TEM), the sarcocyst wall was Type I, with a parasitophorous membrane of approximately 100 nanometers in width arranged in an undulating pattern and intermittently folded inward in a branching pattern. The sarcocysts contained metrocytes in different stages of development and mature bradyzoites. The nucleic acid sequence from a section of the 18S small subunit rRNA gene was most closely related to S. mucosa that uses marsupials as intermediate hosts and has an unknown definitive host. This is apparently the third report of muscular Sarcocystis infection in snakes and is the first to describe the ultrastructure of the sarcocysts and use sequencing methods to aid in identification.

Toxoplasma gondii is a protozoal parasite with worldwide distribution that is able to infect a wide variety of mammals and birds. Our main goal was to screen for T. gondii antibody titers in a previously untested species, the spotted hyena (Crocuta crocuta); however, this goal first required us to investigate serological procedures that could be suitable for hyenas. Cats are the closest domestic relations of hyenas, so T. gondii antibody titers were first compared in 26 feral cats with specific or nonspecific fluorophore-labeled secondary reagents, i.e., anti-cat IgG or protein A. Substitution of anti-cat IgG with protein A caused a statistically significant drop in titer measurements in cats (P = 0.01) with a reduction of the geometric mean titer equivalent to 1 doubling-dilution. The same procedures were then applied to captive spotted hyenas. Titers measured in 9 of 10 hyenas were identical whether anti-cat IgG or protein A was used as the secondary reagent: 5 had titers <1:16, 2 had titers of 1:16, and 2 had titers of 1:32. One hyena had maximum titers of 1:64 or 1:32 when anti-cat IgG or protein A was used, respectively. The use of protein A as the secondary reagent in serologic assays can be applied to a range of mammalian species and seems unlikely to affect test specificity; however, the use of protein A may reduce test sensitivity, as suggested in the present study using cats. Despite a control program, some exposure to T. gondii had occurred in the Zoo's spotted hyenas.