Volume 95 (2009) of the Journal of Parasitology represented a significant benchmark in the history of paleoparasitology when it received on the cover formal recognition as a topical area for publication. This retrospective examination chronicles the emergence of paleoparasitology, from its origins as an adjunct contribution to the study of prehistoric human populations to its modern expression as a sub-disciplinary interest. The aim of paleoparasitology is to elucidate the temporal and spatial dimensions of parasitism from the fossil record of human and non-human host populations.

Avian blood parasites can have significant impacts on adult breeding birds but studies of parasitism outside the breeding season are rare, despite their potentially important implications for host–parasite dynamics. Here we investigate temporal dynamics of blood parasite infection in adult yellowhammers Emberiza citrinella. We screened blood samples collected between December and April of 2 consecutive winters using PCR. We found a high prevalence of both Haemoproteus and Leucocytozoon parasites, with a mean prevalence of 50% across 2 winters. Prevalence of both parasites was higher during the second, colder winter of the study. Temporal trends differed between the 2 genera, suggesting that chronic Haemoproteus infections gradually disappear throughout the winter but that Leucocytozoon infections exhibit a relapse during late winter, possibly coincident with reduced food availability. Our results highlight the difference in temporal dynamics between 2 blood parasite genera infecting the same host population and emphasize the need for accurate assessment of infection status at appropriate time periods when examining impacts of, and associations with, blood parasite infection. We suggest that further research should investigate the implications of over-winter infection for birds' physiology, behavior, and survival.

Examination of extensive Dermacentor Koch, 1844 holdings stored in several major tick collections allowed us to re-evaluate the taxonomic content of Dermacentor everestianus Hirst, 1926 and redescribe all of its parasitic stages in detail for the first time. Examination of the type specimens of Dermacentor abaensis Teng, 1963, a species treated as valid by most workers, and Dermacentor birulai Olenev, 1927, a species some recent authors considered as valid, led us to the conclusion that they are junior synonyms of D. everestianus. The relation of D. everestianus with some other species in the genus is questionable and warrants further studies. From possibly sympatric Dermacentor species, the adults of D. everestianus can be distinguished by the following combination of characters: intensive ivory colored ornamentation of conscutum and scutum, absence of brown patches on lateral fields of conscutum in the male, long and narrow dorsal prolongation of spiracular plates, short cornua, short dorsal spur on trochanter I, and absence of large ventral spur on distal ends of genua and tibiae II–IV. Nymphs of D. everestianus can be distinguished by numerous setae on alloscutum (>48 pairs), large spiracular plates with their longitudinal diameter exceeding that of sclerotized ring around anal valves, moderate lateral projections of basis capituli with blunt apices situated slightly posterior to the midlength of basis capituli dorsally, relatively large auriculae, relatively short, narrowly rounded at apices spurs on coxae I with internal spur being shorter than external and moderate triangular spur on coxae IV; while larvae can surprisingly easily be distinguished from those of other species found in the region by greatly elongated posterior portion of scutum where eyes are situated just posterior to the midlength of scutum. So far, D. everestianus is found only in China and Nepal, where the adults were collected from domestic and wild ungulates while the immature stages were recorded from lagomorphs and rodents.

Thirty-two black-and-red broadbill Cymbirhynchus macrorhynchos were examined for chewing lice in Vietnam. All birds examined were parasitized by Myrsidea claytoni. Mean abundance was 27.3, with intensity range 5–80 lice per bird. This is the first report of a Myrsidea from this host, although the females differ slightly from the original description of M. claytoni from Pycnonotus eutilotus in the shape of metanotum and of tergites I–II. So this is also the first record of 1 species of Myrsidea from 2 very distantly related hosts. Although the original hosts of M. claytoni belong to the family Pycnonotidae, C. macrorhynchos is a member of the family Eurylaimidae, representing the Old World Suboscines, which are considered as a basal lineage among passerines. Therefore, our record represents an interesting case of natural host switching. The high prevalence as well as the intensity of infestation show that M. claytoni is well established on C. macrorhynchos in Vietnam.

The effects of trematode Plagiorchis mutationis parasitism on the cellular immune responses of the snail host Lymnaea stagnalis were investigated. The number of spreading blood cells (hemocytes) from infected snails was significantly less (69.5%) than in uninfected individuals (79.2%). The phagocytic activity of blood cells in infected snails was also significantly less (17.2%) compared to uninfected snails (27.8%). The hemocytes from the infected snails did not form a complete capsule around Sephadex beads in vitro. The protective reactions of the snail hosts were independent of the parasite load (daily cercariae production). In vitro, dead cercariae of P. mutationis were encapsulated by hemocytes from uninfected snails. The hemocytes of the infected snails formed a complete capsule around only 20% of dead cercariae in vitro, with remaining cercariae either unencapsulated (50% of cercariae) or incompletely encapsulated (30% of cercariae). The total number of hemocytes in the infected snails was twofold less than in uninfected individuals. The results of this study showed that the cellular response of snail host L. stagnalis to P. mutationis trematode infection is similar to the previously studied snail–trematode model systems.

Clinical toxoplasmosis has been reported in many species of warm-blooded animals but is rare in camelids. Here we report acute fatal systemic toxoplasmosis involving heart, thyroid gland, stomach, intestine, diaphragm, kidneys, adrenal glands, and liver of a 13-mo-old llama (Llama glama). Many Toxoplasma gondii tachyzoites were associated with tissue necrosis in multiple organs. Death was attributed to severe myocarditis. Ulcers associated with numerous tachyzoites were present in the C3 compartment of the stomach. Tissue cyst development was followed using bradyzoite-specific T. gondii antibodies. Individual intracellular, and groups of 2 or more, bradyzoites were identified in hepatocytes, biliary epithelium, myocardiocytes, lung, diaphragm, thyroid gland, spleen, and stomach. Lesions in the brain were a few microglial nodules and very early tissue cysts containing 1–3 bradyzoites. These observations suggest that the animal had acquired toxoplasmosis recently. Diagnosis was confirmed immunohistochemically by reaction with T. gondii-specific polyclonal rabbit serum but not with antibodies to the related protozoan Neospora caninum. Genetic typing using the DNA extracted from paraffin-embedded myocardium of llama and 10 PCR-restriction fragment length polymorphism (RFLP) markers revealed a type II allele at the SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1 L358, and Apico loci; therefore, this isolate belongs to the ToxoDB PCR-RFLP genotype #1, which is most common in North America and Europe.

Mecistocirrus digitatus is a hematophagous abomasal nematode which causes significant blood and production losses in cattle. The objectives of the present study were to: (1) report the reappearance of M. digitatus in cattle from the Mexican tropics using microscopy, scanning electron microscopy (SEM), and molecular identification, and (2) determine the prevalence of M. digitatus in slaughtered adult cattle from the Mexican tropics. Gastroinestinal nematodes (GIN) were recovered from the abomasum of an 8-yr-old cow (Holstein × Zebu) previously diagnosed with Johne's disease. Of 1,254 GIN, 98.8% were identified as M. digitatus and 1.2% as Haemonchus sp. SEM was used to identify ultrastructure features of M. digitatus (oral cavity, cervical papillae, bursa, bursa lobes papillae, male spicules, anus, and female vulva). A conventional PCR method was used to corroborate the morphological findings. The prevalence of adult cattle infected with M. digitatus and Haemonchus sp., determined from 68 adult cattle from different grazing tropical herds, was 38.2% and 8.8%, respectively. Ninety-eight percent of animals infected with M. digitatus presented lesions in their abomasum such as mucosal inflammation, hemorrhage, and ulcers; some of them had necrosis. The current reappearance of M. digitatus in a Mexican herd suggests the possibility of an underestimated prevalence of this nematode amongst grazing cattle.

Five of 16 (31%) rufous-capped warblers (Basileuterus rufifrons) and 2 of 5 (40%) ovenbirds (Seiurus aurocapilla) sampled from Costa Rica were positive for a novel species of Isospora. Oocysts have a thin, smooth, double-layered, colorless wall and measure 22.3 μm ± 1.6 μm × 24.3 μm ± 1.5 μm (19–25 μm × 21–28 μm) with an average length-width (L/W) ratio of 1.0 (1–1.3). Oocyst residuum and micropyle are absent, but 0–4 spherical to cigar-shaped polar granules (1–2.5 μm) are present. Sporocysts are ovoid and measure 11.8 μm ± 0.9 μm × 16 μm ± 1.7 μm (10–14 μm × 12–19 μm) with an average L/W ratio of 1.6 (1.0–1.9). A knob-like Stieda body continuous with the sporocyst wall and a trapezoidal compartmentalized substieda body are present. Each sporocyst contained 4 sporozoites and a diffuse sporocyst residuum consisting of many variable-sized granules, some as large as 2 μm. This is the second description of an Isospora species in New World warblers (Passeriformes: Parulidae) and the first report of Isospora from both the rufous-capped warbler and ovenbird.

Monotypic Paracardicoloides Martin, 1974 is emended based on supplemental observations of 2 voucher specimens of Paracardicoloides yamagutii Martin, 1974. Features of the anterior sucker, esophagus, and intestine as well as the male and female genitalia previously attributed to, or omitted from the diagnosis and description of, this fluke are resolved and further detailed herein. The holotype of P. yamagutii, originally deposited in the Allan Hancock Parasitology Collection, apparently has been lost, and Queensland Museum voucher G222650 is designated as the neotype. Elopicola nolancribbi n. gen., n. sp. infects the ladyfish, Elops saurus, (Elopiformes: Elopidae) in the north-central Gulf of Mexico. The new genus resembles Paracardicoloides by having the combination of a bowl-shaped anterior sucker, inverse U-shaped intestine, short posterior ceca, post-cecal ovary, pre-ovarian ootype, compact uterus, and prominent excretory arms and vesicle. It differs from Paracardicoloides by lacking robust tegumental body spines and by having a single testis, post-testicular ovary, and primarily post-testicular uterus. The present study is the first confirmed report of an aporocotylid infecting a member of Elopiformes as well as the first report of an infection in an elopomorph outside of Australia and New Zealand or in marine waters.

Spauligodon bintangensis n. sp. from the intestines of Cyrtodactylus bintangrendah (Gekkonidae) from Peninsular Malaysia is described and illustrated. Spauligodon bintangensis n. sp. represents the 51st species assigned to the genus and the first species from the Oriental Region. The new species is most similar to Spauligodon atlanticus, Spauligodon eremiasi, and Spauligodon occidentalis, but is easily separated by position of vulva, prebulbar in S. atlanticus and S. occidentalis, postbulbar in the new species, and location of lateral alae; in S. eremiasi, the lateral alae occur only in the fourth quarter of the body, whereas in the new species the lateral alae begin just posterior to lips.

Two morphologically disparate undescribed species of diphyllidean cestodes from the Yellowspotted skate, Leucoraja wallacei, from South Africa were included in a recent molecular phylogenetic study aimed at revising diphyllidean classification. From a molecular standpoint, these species were determined to be only distantly related to one another. One (originally referred to as Echinobothrium n. sp. 2) showed affinities with species of the batoid-parasitizing Echinobothrium sensu stricto and is described here as Echinobothrium marquesi n. sp. This species most closely resembles Echinobothrium joshuai, an affinity supported by the previous molecular study, but differs in the form of its “B” hooks and degree of overlap between bothria and cephalic peduncle. The other species (originally referred to as New genus n. sp. 1), although exhibiting the full complement of scolex armature, grouped among primarily shark-hosted genera, most of which lack or exhibit reduced scolex armatures. That unexpected result suggested that erection of a novel genus might be warranted, but morphological grounds supporting the action were not apparent at that time. The present study aimed to explore the morphology of this taxon in more detail. Light and scanning electron microscopy revealed this taxon to be exceptional in that its lateral hooklets, which are arranged in 2 clusters like those of Echinobothrium and Coronocestus, are unique in being arranged in anterior and posterior rows, rather than in a single row. Andocadoncum n. gen. is erected, with Andocadoncum meganae n. sp. as its type, to accommodate this taxon. A minor adjustment to the existing hook formula by presenting counts for the anterior hooklets (a) separately from the posterior hooklets (b) readily accommodates this new form. Leucoraja wallacei is distinctive among skates in hosting 2 different genera of diphyllideans. In the context of the previous molecular phylogenetic analyses, it appears that the presence of Andocadoncum n. gen. in this skate species may be the result of a host-switching event involving a weakly or unarmed shark-hosted taxon moving to a batoid—a shift that was likely accompanied by a return to the fully armed condition seen, for example, in other batoid-parasitizing members of the order.

Auriculostoma totonacapanensis n. sp. is described from the Mexican tetra, Astyanax mexicanus (Actinopterygii, Characidae) collected in a tributary creek of the Bobos River in Filipinas, Veracruz, Mexico. The new species is set apart from all congeners by the combination of some morphological characters such as the testes position (oblique in most specimens), the ventral to oral sucker ratio (1:1.2 × 1:1.1), the cirrus sac originating at the ovarian region, and by having vitelline follicles not confluent in the posttesticular region. Auriculostoma totonacapanensis n. sp. closely resembles Auriculostoma platense (Szidat, 1954) Scholz, Aguirre-Macedo, and Choudhury, 2004 and Auriculostoma diagonale Curran, Tkach and Overstreet, 2011 by possessing oblique testes; however, it differs from both species by possessing a genital pore located at level of the cecal bifurcation and by having vitelline follicles extending anteriorly up to the cecal bifurcation level, instead of a genital pore located between the anterior margin of the ventral sucker and cecal bifurcation, and vitelline follicles extending anteriorly to the mid level of the esophagus as in A. platense or to the pharynx level as in A. diagonale. Additionally, the new species differs from A. diagonale by having vitelline follicles not confluent or scarcely confluent in the posttesticular region rather than extensively confluent. Scanning electron microscopy micrographs of the new species demonstrated the presence of a single pair of muscular lobes on either side of the oral sucker, with a broad base, stretching from the ventrolateral to the dorsolateral side. Maximum parsimony and Bayesian inference analyses of the 28S rRNA gene sequences placed A. totonacapanensis as sister species of Auriculostoma astyanace Scholz, Aguirre-Macedo, and Choudhury, 2004. Nucleotide variation between A. totonacapanensis and A. astyanace was 2.0% and 3.6% for the 28S rRNA gene and ITS2 sequences, respectively. Sequence variation for the 28S rRNA gene between Auriculostoma spp. and 7 other genera of Allocreadiidae ranged from 2.4 to 6.3%.

During a survey of blood parasites in a population of Leach's and black storm petrels (Oceanodroma leucorhoa and Oceanodroma melania) in Mexico, infection by a Hepatozoon species in erythrocytes of several birds was noted. Here we describe the species as Hepatozoon peircei sp. nov. Some species of Hepatozoon described from birds have been identified as lankesterellids when DNA molecular analyses were conducted. However, a sequence of 1,774 bp of the parasite found infecting storm petrels in this study clearly show the parasite is a species of the genus Hepatozoon. This is the first Hepatozoon species infecting birds to be characterized at the molecular level and the first found infecting erythrocytes and not leucocytes.

Two new species, Encotyllabe cheilodactyli and Encotyllabe antofagastensis (Monogenea: Capsalidae), have been found in the pharyngeal plates of Cheilodactylus variegatus and Anisotremus scapularis, respectively, in northern Chile (23°38′N, 70°24′W). Descriptions of the new species were based on morphometric and molecular evidence. Both species differ from previously described species of the genus by a combination of characteristics, including the size and relative position of the testes and the shapes of the major and small hamulus. In addition, E. cheilodactyli is unique among the known species of Encotyllabe due to the testes crossing the equatorial line of the body proper, and E. antofagastensis is unique among the known species of Encotyllabe due to its penis structure and anatomy leading to the genital pore. The analysis of the cytochrome c oxidase I gene as well as morphometric analyses demonstrated that the specimens belonged to 2 different species.

Paragyrodactylus variegatus n. sp. (Monogenea: Gyrodactylidae) is described from the fins and body surface of Homatula variegata (Dabry de Thiersant, 1874) living in cold-water streams in the Qinling Mountains of central China. It represents the third known species in the genus, with all species parasitizing Asian river loaches (Cypriniformes: Nemacheilidae). The new species resembles most closely Paragyrodactylus iliensis Gvosdev and Martechov, 1953, but is diagnosed by its marginal hooks being longer than 19 μm, a sickle longer than 4 μm, a sickle aperture shorter than the sickle distal width, and absence of a ventromedial indentation in the haptoral accessory hard part. Scanning electron microscopy reveals a central foramen in the haptoral accessory hard part, and small knob-like protuberances on the heel and toe of the marginal hook sickle. A partial 18S (448 bp) and complete ITS-1, 5.8S, and ITS-2 (1,139 bp) rDNA sequences are provided. A preliminary phylogenetic analysis based on the partial 18S gene, which included all sequenced genera of the Gyrodactylidae, suggests P. variegatus n. sp. is basal within Gyrodactylus sensu lato. It is proposed that the genus is a relict freshwater lineage parasitizing river loaches in the mountain plateaus of central Asia.

Ancient DNA (aDNA) of Encephalitozoon intestinalis (Microsporidia, Fungi) was detected in archaeological material originated from New Town of Prague (Czech Republic) with the use of molecular methods. Microsporidial aDNA was found in 3 samples originating from 2 objects, in a well/cesspit (samples from layers from the 18th century) and in a well from the 18th/19th century. The ability to use molecular methods to detect microsporidia extends the range of paleoparasitological inquiry, and could contribute to a better understanding of parasites shared between human and animals.

Trichomonas gallinae , the cause of avian trichomonosis, is most commonly found in the order Columbiformes. Racing pigeons are often treated preventively with nitro-imidazoles, which could result in the emergence of resistant isolates, and these isolates can be a threat to wildlife when exchanges occur. The sequence type of 16 T. gallinae isolates obtained from racing pigeons and 15 isolates from wild pigeons was determined based on the ITS1/5.8S rRNA/ITS2 region sequence. In addition, the resistance profiles of these isolates against 5 different nitro-imidazoles (metronidazole, dimetridazole, ronidazole, tinidazole, and carnidazole) were determined. Two different Trichomonas sequence types were isolated. Sequence type A isolates were recovered from racing and wild pigeons, in contrast to sequence type B, which was only isolated from wild pigeons. Isolates with sequence type B were all susceptible to the tested nitro-imidazoles, except for tinidazole resistance in 3 isolates. Resistance to the nitro-imidazoles was observed more frequently in isolates obtained from racing pigeons than from wild pigeons, with most isolates belonging to sequence type A. A higher percentage of the sequence type A isolated from racing pigeons, in comparison with those isolated from the wild pigeons, were resistant to the nitro-imidazoles and displayed higher mean lethal concentration (MLC) values. Two isolates belonging to sequence type A, 1 recovered from a racing pigeon and 1 from a wild pigeon, displayed a similar resistance pattern, suggesting a potential exchange of resistant isolates between racing pigeons and wild pigeons.

The identification of excreted–secreted (ES) proteins of filarial nematodes as potential diagnostic reagents is an important requirement for the development of methods to determine level of infection in the host, especially for human filariae. Dirofilaria immitis, the canine heartworm, is a widespread and important veterinary pathogen and is a useful model for filarial parasites of humans. An analysis of proteins released from adult D. immitis (the secretome) in culture is available. We sought to identify D. immitis ES proteins found in vivo to validate the in vitro secretome and to investigate them as potential diagnostic reagents. Cultures of D. immitis adults obtained from infected dogs were maintained for 72 hr with daily changes of media. Proteins were concentrated from spent media by standard methods and were passed through Protein-A columns containing purified IgG antibodies from heartworm-infected dogs. Following extensive washing, heartworm proteins recognized by the antibodies were eluted from these columns and submitted for analysis by tandem mass spectrometry (MS/MS). As a comparison, somatic proteins from adult D. immitis female parasites and microfilaria were also processed and analyzed by the same protocol. Six, 9, and 12 proteins were identified by MS/MS in the ES, adult female, and microfilaria samples, respectively. The identification of the most abundant parasite proteins present in the serum of infected hosts offers a rational approach to the development of new diagnostic assays that may be applicable across the Filarioidea.

Many species of fish in the world are infected with digenean trematodes belonging to the genera Clinostomum and Euclinostomum. In this study, metacercariae, identified as Euclinostomum sp. on the basis of morphological characteristics and molecular data, were taken from 3 osphronemid fish—Trichopsis vittata, Trichopsis schalleri, and Betta imbellis, in Thailand. Phylogenetic analysis based on a mitochondrial gene (cytochrome c oxidase subunit I) and 2 nuclear genes (18S rDNA and ITS—internal transcribed spacer) of these Euclinostomum parasites indicated a clear distinction from those belonging to the Clinostomum genus. These are the first records of partial mitochondrial and nuclear DNA sequences of an Euclinostomum sp.