Since the late 19th century, the ectoparasitic copepod Salmincola cf. edwardsii has been observed infecting brook trout (Salvelinus fontinalis) in Wisconsin streams. A perceived increase in the incidence of S. cf. edwardsii infection across the state and the observation of an S. edwardsii epizootic leading to declines in brook trout recruitment in Ash Creek, Wisconsin, have raised concerns about the extent to which S. edwardsii threaten other populations of brook trout in Wisconsin streams. In 2013–2017, brook trout were inspected in 282 streams across the state to determine the distribution, prevalence, and maximum intensity of infection of adult female copepods whose morphology was consistent with S. edwardsii. Salmincola cf. edwardsii were present in 79% of streams and absent from 21%. Prevalence of infection ranged from 0.4 to 100% where the parasite was present. Maximum intensity of infection was low (1–5 S. cf. edwardsii) in 31% of streams, moderate (6–14) in 35% of the streams, and high (≥15) in 34% of the streams. Maximum intensity was high and prevalence exceeded 90% of brook trout in 9 streams, suggesting epizootics as observed in Ash Creek are uncommon. Our statewide distributional data are critical to understanding the scope of S. cf. edwardsii infection of brook trout in Wisconsin's recreational fisheries and will provide a baseline for ongoing and future investigations of S. cf. edwardsii–brook trout dynamics.

Cryptosporidiosis is one of the leading causes of diarrhea in humans and several other vertebrate species. Because surveys of Cryptosporidium genotypes from animals and humans living in the same region are rare, our understanding of the importance of zoonotic transmission in the epidemiology of cryptosporidiosis remains superficial. PCR was used to amplify a portion of the Cryptosporidium 18S small subunit ribosomal RNA gene from fecal DNA from humans and livestock living in Ekiti and Oyo states, Nigeria. PCR-positive samples were further analyzed using PCR targeting the heat-shock protein HSP-70, the actin, and the sporozoite glycoprotein gene gp60. A questionnaire was used to collect demographic information. Sixteen of 187 samples collected were Cryptosporidium 18S PCR positive. Of these, 5 samples originating from HIV-positive patients, 5 from otherwise healthy children, 2 from chickens, 3 from goats, and 1 from a dog were positive for at least 1 marker. Sequencing of the 18S rRNA amplicons revealed the presence of Cryptosporidium parvum in 2 HIV positive patients and in a child; the actin sequence confirmed the presence of this species. Two samples of HIV-positive patients amplified Cryptosporidium hominis 18S rRNA, one of them confirmed by the HSP-70, actin, and gp60 sequences. Cryptosporidium meleagridis was found in another HIV patient, while C. hominis was detected in 3 children (of which 2 were confirmed by gp60). Cryptosporidium muris was found in 1 child. In birds, we found C. meleagridis and, significantly, C. parvum, whereas we detected C. parvum and C. muris in 1 goat each. The only dog sampled was positive for Cryptosporidium canis. We conclude that, in the environment we surveyed, humans and animals are a potential part of the same transmission cycle. Measures to prevent zoonotic transmission should therefore be considered to reduce the prevalence of cryptosporidiosis.

The distribution of the freshwater myxozoan parasite Ceratonova shasta in the Pacific Northwest of North America is limited to overlap in the ranges of its 2 hosts: the polychaete Manyunkia sp., and Pacific salmonids. Studies in the Klamath River (Oregon/California) and Deschutes River (Oregon), showed that the parasite population is comprised of multiple sympatric genotypes, some of which correlate with particular salmonid host species and with differences in clinical disease in those hosts. The 3 primary genotypes O, I, and II are defined by the number of a specific tri-nucleotide repeat in the internal transcribed spacer-1 region. To understand the spatial extent of host–parasite genotype patterns, we sequenced the parasite from 448 salmonid fishes from river basins in California, Oregon, Washington, Idaho, and British Columbia, Canada. We sampled intestinal tissues from 6 species of salmon and trout, both those that exist naturally with the parasite (sympatric) and those that do not naturally co-occur with the parasite and were exposed artificially in cages (allopatric). In most river basins we detected the same primary C. shasta genotypes that were described from the Klamath and Deschutes rivers, and we did not detect any novel primary genotypes. Host–parasite genotype patterns were consistent with previous data: genotype O was found in sympatric trout only; genotype I predominantly in Chinook salmon, and genotype II in all 6 fish species but dominant in coho salmon. Our findings of widespread, consistent host–parasite genotype patterns support the hypothesis that C. shasta has a long evolutionary history with salmonid fishes in the Pacific Northwest, and impels additional studies to determine if these parasite genotypes should be considered different species.

Diagnosis of parasitic diseases that involve tissue-stage larvae is challenging, and serology remains the most effective antemortem test for detecting these infections. Baylisascaris procyonis, the raccoon roundworm, is a zoonotic ascarid. Raccoons are the usual definitive host, and humans may be infected as accidental hosts. More than 150 species of birds and mammals may act as paratenic hosts, and rodents play an important role in the transmission and maintenance of this parasite in nature. Migratory larvae in paratenic host tissues can produce ocular disease and severe to fatal neurologic disease, but not all infected hosts develop signs. A sensitive and specific Western blot (WB) assay based on a recombinant Baylisascaris-specific antigen (rBpRAG-1) has been developed for use in humans. We evaluated the use of this antigen to detect Baylisascaris spp. infections in rodent paratenic hosts. With the use of 4 species of Peromyscus mice (Peromyscus californicus, Peromyscus leucopus, Peromyscus maniculatus, Peromyscus polionotus) from a previous infection trial, we developed species-adapted WB and ELISA assays and evaluated performance compared to detection of larvae in tissue samples. These assays revealed species-level differences in seroconversion and terminal antibody concentrations, with P. leucopus developing significantly greater antibody concentrations than P. californicus and P. polionotus at all dose levels, and P. maniculatus at the low dose. Some P. californicus and P. polionotus failed to seroconvert despite the recovery of larvae from their tissues. WB and ELISA results were correlated; however, the WB demonstrated higher sensitivity than the ELISA overall (72.2% versus 63.9%, respectively). With the use of experimental samples, specificity was 100% for WB and 94.1% for ELISA. A WB was also used to test Mus and Rattus samples, and although numbers were too limited to evaluate sensitivity and specificity, all animals known to be infected by tissue digestion were WB positive, and all uninfected animals were negative. Finally, the Peromyscus-adapted WB and ELISA were used to test a set of serum samples from wild-trapped P. maniculatus and Rattus rattus. Both assays were generally sensitive, but specificity was equivocal. This emphasizes the challenge of using serology for investigation of wildlife diseases, in which hosts have unknown exposure histories. Nevertheless, serologic methods have utility in the study of Baylisascaris spp. in paratenic hosts, either wild or captive, and have advantageous attributes (non-lethal, high-throughput), but results should be interpreted carefully.

Trematodes of the family Dicrocoeliidae commonly use terrestrial mollusks as the first intermediate host. Despite abundant studies on the adult worms in birds and mammals, few reports exist on their larval stage in snail intermediate hosts. A present survey of mollusks in Japan led us to the discovery of dicrocoeliid sporocysts with cercariae in 16 of 303 individuals, encompassing 8 snail species and 1 slug species. A DNA barcoding based on sequencing of mitochondrial cytochrome c oxidase subunit 1 showed that the larvae consisted of 5 species. Phylogenetic trees of nuclear 18S and 28S ribosomal DNAs confirmed the 5 species to be members of the Dicrocoeliidae. These were temporarily termed dicrocoeliid species 1 to 5, because conclusive identification was impossible without adult worms. These unknown species were phylogenetically related to each other, except for sp. 5. The phylogenetic trees demonstrated close genetic relationships between sp. 3 and the genus Lutztrema and between sp. 5 and the genus Lyperosomum. The phylogenetic analysis also suggests a possibility that the currently accepted macrotaxonomy of the Dicrocoeliidae is problematic, due to the paraphyly of the subfamilies Dicrocoeliinae and Leipertrematinae. Morphological characterization of the cercariae and their DNA barcodes provide a primary platform for differentiating dicrocoeliids from various mollusks in Japan. The DNA barcodes, in particular, will enable tracing the parasite life cycles, in case of finding metacercariae and adults from presently unknown hosts.

Neoechinorhynchus bullocki n. sp. is described from Catostomus commersonii Lacépède, 1803, from the Otsego Lake watershed, central New York State. Neoechinorhynchus bullocki differs from other catostomid-infecting species in body size and shape, proboscis armature, and characteristics of the male reproductive system. Sequences of the internal transcribed spacer and the large ribosomal subunit (28S) were also used to distinguish this new species. Neoechinorhynchus bullocki is the sixth species of Neoechinorhynchus to be reported from C. commersonii and the seventh species of the genus with a dorsal body wall considerably thicker than the ventral body wall. It is the 34th species of Neoechinorhynchus to be described from fishes of North America. In contrast to many of its congeners, N. bullocki was found to have a strict (i.e., oioxenous) level of host specificity based on an extensive fish parasite survey that took place in New York State over a 10-yr period and included the examination of the intestines of more than 1,800 fish from the fish families Catostomidae, Centrarchidae, Cyprinidae, Esocidae, Ictaluridae, and Percidae.

Specimens of the African snail Achatina fulica, collected in Bucaramanga, Colombia, were examined for parasites. Numerous specimens of Caenorhabditis briggsae were collected from the digestive tract of the snails and identified by the structure of male spiculum, caudal bursa, gubernaculum and precloacal lip in males, triangular tooth in metarhabdion, and protandrous hermaphrodites with a female:male ratio of 15:1 and with morphometry. DNA sequences of the ITS2 region of the ribosomal gene array from worms in this study matched with 99% similarity to published sequences of C. briggsae. A redescription of the species is provided. This is the first record of the species in South America.

Species of the eutetrarhynchid genus Hispidorhynchus Schaeffner and Beveridge, 2012 possess an uncinate macrohook on the bothrial surface of the basal swelling of each tentacle. This unique feature of the oncotaxy is only shared with the closely related genus Oncomegas Dollfus, 1929. A new species of Hispidorhynchus is described from specimens infecting Styracura schmardae (Werner, 1904) (Potamotrygonidae) from the western Caribbean Sea off the coast of Belize and Panama. Hispidorhynchus styracurae n. sp. differs from its 3 congeners in the possession of smaller and narrower bulbs, fewer principle hooks in the metabasal armature, size of the macrohook, and different scolex proportions. Scanning electron microscopy reveals new information on the microthrix morphology of the genus. A diagnostic key for the differentiation of species of Hispidorhynchus is provided. In addition, new host and locality records are reported for Oncomegas wageneri (Linton, 1890) Dollfus, 1929, collected from Hypanus guttatus (Bloch and Schneider, 1801) (Dasyatidae) off the coast of Maceió, Alagoas (Brazil). This considerably expands the geographical distribution of this species in the western Atlantic Ocean.

Dirofilaria immitis is a common filarial parasite found in dogs and cats in the Americas, with the pathophysiological consequences of the infection differing somewhat between these 2 host species. Recent research efforts have been focused on determining if the microRNAs (miRNAs) released from adult Dirofilariae have a role as markers for distinguishing the intensity of adult worm infection, as well as determining the presence of new infections. This study expands previous work on 2 nematode miRNAs, miR34 and miR-71, by addressing their ability to discriminate between low and high D. immitis adult worm intensities in dogs. Serum samples were collected from 13 dogs, 8 of which carried known numbers of adult D. immitis at autopsy in their hearts and pulmonary vessels. Three groups of canine sera were created based on D. immitis burden: “control” (0 worms; 5 animals), “low intensity” (10–18 worms; mean ± SD = 12.3 ± 4.4; 4 animals), and “high intensity” (41–72 worms; mean 62.5 ± 15.1; 4 animals) groups. A qPCR analysis was performed on each sample to measure plasma levels of miR-34 and miR-71; however, no significant differences were observed between these groups in terms of levels of miRNAs, so the low- and high-intensity samples were then combined into a single “infected” category and compared to the “non-infected” controls. Copy numbers of both miR-34 and miR-71 were significantly higher in infected compared to uninfected animals (P = 0.015 and P = 0.027, respectively). The Ct values of expression compared with the adult worm intensity for each miRNA revealed that both miR-34 and miR-71 significantly discriminate between the infected and non-infected groups (P value < 0.0001 for both). These findings support the contention that miRNA 34 and miRNA 71, which are filarial-specific miRNAs, can both serve as biomarkers for the presence of D. immitis infection in dogs, but at this point they do not appear to reflect the actual intensity of adult parasites present.

Giardia intestinalis is a parasite that commonly causes diarrheal disease throughout the world. An accurate and rapid diagnosis is essential to reduce the infection. Classical diagnosis of giardiasis is performed by microscopic examination of stool samples, but in recent years many DNA-based methods have been developed. In this preliminary observational study, we compared the results of the commercial BD Max enteric parasite panel (EPP) with an in-house real-time (Rt) PCR for G. intestinalis. The study population was composed of 73 samples. Of these, 27 tested positive with both techniques and 39 tested negative. Seven samples were positive with the in-house Rt PCR and negative with the BD Max EPP. The Cohen's kappa was 0.805 (95% CI 0.670–0.940). In conclusion, these preliminary results suggest that the Rt-PCR could possibly demonstrate higher sensitivity for the diagnosis of G. intestinalis than BD Max EPP, which tended to miss infection of low intensity.

In the course of a structure-based drug discovery program the known anticancer candidate marimastat was uncovered as a potent inhibitor of an enzyme in nematode cuticle biogenesis. It was shown to kill Caenorhabditis elegans, and the sheep parasites Haemonchus contortus and Teladorsagia circumcinta via an entirely novel nematode-specific pathway, specifically by inhibiting cuticle-remodeling enzymes that the parasites require for the developmentally essential molting process. This discovery prompted an investigation of the compound's effect on Heligmosomoides polygyrus parasites in a mouse model of helminth infection. Mice were administered the drug via oral gavage daily from day of infection for a period of 2 wk. A second group received the drug via intra-peritoneal implantation of an osmotic minipump for 4 wk. Control groups were administered identical volumes of water by oral gavage in both cases. Counts of H. polygyrus fecal egg and larval load showed that marimastat effected a consistent and significant reduction in egg laying, and a consistent but minor reduction in adult worm load when administered every day, starting on the first day of infection. However, the drug failed to have any significant effect on egg counts or worm burdens when administered to mice with established infections. Therefore, marimastat does not appear to show promise as an anthelmintic in gastrointestinal nematode infections, although other metalloproteases such as batimastat may prove more effective.

The protozoan parasite Sarcocystis falcatula is an important cause of clinical disease in several avian intermediate hosts. The host range of S. falcatula is wide, and numerous outbreaks of acute sarcocystosis have been reported in passerine and psittacine birds in captivity in the Americas. Previous diagnosis was performed by serologic methods, light, and/or electron microscopic examinations with limited molecular confirmation. Here, we report histological and molecular diagnosis of acute, fatal S. falcatula infections in rainbow lorikeets (Trichoglossus moluccanus) at the Philadelphia Zoo. Pulmonary sarcocystosis was suspected antemortem in 3 lorikeets (3–5 yr old); these birds died despite antiprotozoal therapy. The predominant lesion was pneumonia associated with S. falcatula–like schizonts in pulmonary vascular endothelium. The multilocus PCR-DNA sequencing (18S rDNA, 28S rDNA, ITS-1, and cox1) of frozen lung tissue confirmed S. falcatula infections in all 3 birds. Our results and previous studies suggest that acute pulmonary form of sarcocystosis is a major contributor of death to Old World psittacine birds.

Litomosoides spp. (Nematoda: Filarioidea: Onchocercidae) are of great interest for understanding dynamics of zoonoses because of their similarity to human-infecting filariae (e.g., Onchocerca volvulus). We encountered Litomosoides microfilaria in blood samples from 7 Panamanian bat species. Species were identified through morphological analysis. We assessed individual traits and immune response of the host individuals as well as host species ecology. We found 8 host-specific species/morphotypes of Litomosoides microfilariae in 5 bat species, with 5 individuals showing multiple infections. Our study includes the first records of Litomosoides brasiliensis in Artibeus jamaicensis and of Litomosoides-like and Litomosa-like microfilaria in Lophostoma silvicolum. Multiple binary regression analysis showed that the presence of specific Litomosoides species was influenced by host species, sex, and reproductive status.

Knowledge regarding endoparasites of Galapagos sea lions, Zalophus wollebaeki, is limited to 1 report. Herein, we examined feces extracted from the lower gastrointestinal tract of 15 Galapagos sea lions plus 14 fecal mounds voided by Galapagos sea lions at 4 locations on San Cristobal Island, Galapagos in May and June of 2016. With the use of standard fecal flotation and sedimentation techniques, lungworm larvae suggestive of Parafilaroides and Otostrongylus sp., eggs of pseudophyllidea cestodes and anisakid nematodes, and coccidian oocysts were collected from study samples. This is the first report of potential lungworm larvae, anisakids, pseudophyllidean cestodes, and coccidian parasites in Galapagos sea lions and demonstrates the importance of fecal survey techniques in describing patterns of parasitism in endangered or protected host populations.

Entomopathogenic nematodes are excellent organisms for dissecting the molecular basis of parasitism and probing the insect innate immune system. The nematode parasite Steinernema carpocapsae is a potent pathogen of insects that has emerged recently as a model for parasitic infection and anti-nematode immune signaling and response. The nematodes are mutualistically associated with the bacteria Xenorhabdus nematophila, which are also pathogenic to insects. Separation of nematodes from their associated bacteria facilitates mechanistic studies focusing on the impact of the parasites without considering the contribution of their bacterial partners. An important aspect in insect infection experiments with entomopathogenic nematodes includes the storage duration of the parasites. Here we have infected larvae of the model insect Drosophila melanogaster with S. carpocapsae nematodes that had been stored for 3 wk or 3 mo. Survival data consistently revealed that infective juveniles with prolonged storage exhibit substantially increased virulence toward D. melanogaster larvae compared with those that had been stored for a shorter time, and the presence of mutualistic X. nematophila in the nematodes does not influence this result. Although the basis for this effect is currently unknown, these surprising findings indicate that prolonged nematode storage can markedly alter virulence. This is significant knowledge that should be taken into account in functional assays involving infection with parasitic nematodes. Future efforts will focus on the identification and characterization of the factors that might determine the interrelationship between prolonged storage and virulence in nematode parasites.

Theileriosis is an important tick-borne pathogen of livestock globally, causing severe reduction of livestock productivity and economic loss. To systematically investigate the prevalence and species of Theileria spp. in goats from Shaanxi Province, a total of 509 blood samples were collected from dairy, cashmere, and meat goats from 7 counties, and examined by using a microscope and the nested PCR targeting the SSU rRNA gene. Of them, 268 (52.7%, 268/509) were positive for Theileria infection. The prevalence was closely associated with ages and production categories. The highest infection was found in meat goats of 7 to 12 mo, and lowest was detected in cashmere goats of 3 to 6 mo. Sequence analysis indicated the presence of 2 Theileria species, with Theileria luwenshuni as the prevalent species and the first report of Theileria sp. OT3 in goats in China. These findings indicate the wide distribution of Theileria spp. in goats of Shaanxi Province and shed new light on the distribution of this parasite in China.