The Indonesian island of Sulawesi is a globally significant biodiversity hotspot with substantial undescribed biota, particularly blood-borne parasites of endemic wildlife. Documenting the blood parasites of Sulawesi's murine rodents is the first fundamental step towards the discovery of pathogens likely to be of concern for the health and conservation of Sulawesi's endemic murines. We screened liver samples from 441 specimens belonging to 20 different species of murine rodents from 2 mountain ranges on Sulawesi, using polymerase chin reaction (PCR) primers targeting the conserved 18S rDNA region across the protozoan class Kinetoplastea. We detected infections in 156 specimens (10 host species) with a mean prevalence of 35.4% (95% confidence interval [CI] = 30.9–39.8%). Sequences from these samples identified 4 infections to the genus Parabodo, 1 to Blechomonas, and the remaining 151 to the genus Trypanosoma. Within Trypanosoma, we recovered 17 haplotypes nested within the Trypanosoma theileri clade infecting 117 specimens (8 host species) and 4 haplotypes nested within the Trypanosoma lewisi clade infecting 34 specimens (6 host species). Haplotypes within the T. theileri clade were related to regional Indo-Australian endemic trypanosomes, displayed geographic structuring but with evidence of long-term connectivity between mountains, and had substantial phylogenetic diversity. These results suggest T. theileri clade parasites are native to Sulawesi. Conversely, T. lewisi clade haplotypes were recovered from both endemic and introduced rodents, demonstrated complete geographic separation between clades, and had low genetic diversity. These results suggest that the T. lewisi clade parasites invaded Sulawesi recently and likely in 2 separate invasion events. Our results provide the first records of metakinetoplastids in Sulawesi's rodents and highlight the need for more extensive sampling for pathogens in this biodiversity hotspot.

The genus LangeroniaCaballero and Bravo-Hollis, 1949, currently contains 6 species of amphibian trematodes distributed in North and Middle America. The type species of the genus, Langeronia macrocirraCaballero and Bravo-Hollis, 1949, occurs in Mexico and is relatively commonly found as a parasite of leopard frogs. However, information regarding its life cycle is lacking. In this paper, we study the life cycle of L. macrocirra in Laguna Escondida, Los Tuxtlas, Veracruz. Definitive hosts (Rana spp.) as well as potential intermediate hosts (gastropods, bivalves, crustaceans, tadpoles, hemipterans, and odonate naiads) were sampled in the locality and studied to search for the presence of adults and larval stages of the trematode. Specimens were morphologically characterized, and some individuals were sequenced for 1 ribosomal gene (28S rRNA) and 1 mitochondrial gene (COI). DNA sequences of the adults obtained from leopard frogs were matched with those of the larval forms in their intermediate hosts (metacercariae, cercariae, and sporocysts) to demonstrate conspecificity. Further, we conducted a detailed study of the tegument of the body surface with scanning electron microscopy to characterize each of the developmental stages of the life cycle of L. macrocirra.

Ticks and tick-borne diseases are important issues worldwide because of their effects on animal and human health. The genus Ornithodoros, which is included in the family Argasidae, is typically associated with wild animals, including seabirds. In this study, samples from the nests of seabirds and surrounding soil were collected to investigate Ornithodoros spp. from 9 uninhabited islands in the western, eastern, and southern parts of Korea from April 2017 to October 2018. The islands are known as the breeding places of migratory and resident birds. Ticks were collected from soil and nest material of seabirds using a Tullgren funnel and identified using 16S rRNA and the cytochrome c oxidase 1 gene (COI), and host animals of soft ticks were identified using the mitochondrial DNA cytochrome b gene by a polymerase chain reaction. In the sequence identity of the 16S rRNA gene fragment of Ornithodoros sp., Ornithodoros sawaii was identified as the closest homologous sequence, and the new Ornithodoros sp. was newly identified. We found that the newly identified Ornithodoros sp. in the Republic of Korea was located in uninhabited islands used as breeding places by the black-tailed gull, Larus crassirostris.

Peroxisome proliferator–activated receptor gamma (PPARγ) regulates neuroinflammation, and its agonists act as neuroprotective agents. This study aims to investigate the correlation between PPARγ and proinflammatory enzyme expression in astroglia infected with Toxoplasma gondii tachyzoite in vitro. Our results showed that matrix metalloprotease (MMP)-2, MMP-9, cyclooxygenase-2 (COX-2), prostaglandin (PGE)-2, inducible nitric-oxide synthase (iNOS), and nitric oxide (NO) were significantly increased in T. gondii–infected astroglia. Furthermore, the expression levels of MMP-2, MMP-9, COX-2, PGE-2, iNOS, and NO were significantly decreased by rosiglitazone—a PPARγ agonist. By contrast, the treatment with GW9662, a PPARγ antagonist, efficiently increased the expression levels of MMP-2, MMP-9, COX-2, PGE-2, iNOS, and NO. These results suggested that the treatment with rosiglitazone offers a potential strategy for controlling the inflammatory factors in T. gondii infection.

Anisakis simplex, Pseudoterranova decipiens, and Contracaecum osculatum third-stage larvae (L3) are fish-borne nematodes that can cause human anisakidosis. Although A. simplex is a known source of allergens, knowledge about the allergic potential of P. decipiens and C. osculatum is limited. Therefore, we performed comparative proteomic profiling of A. simplex, P. decipiens, and C. osculatum L3 larvae using liquid chromatography–tandem mass spectrometry. In total, 645, 397, and 261 proteins were detected in A. simplex, P. decipiens, and C. osculatum L3 larvae, respectively. Western blot analysis confirmed the cross-reactivity of anti-A. simplex immunoglobulin (Ig)G antibodies with protein extracts from P. decipiens and C. osculatum L3 larvae. The identified proteins of the Anisakidae proteomes were characterized by label-free quantification and functional analysis, and proteins involved in many essential biological mechanisms, such as parasite survival, were identified. In the proteome of A. simplex 14, the following allergens were identified: Ani s 1, Ani s 2 (2 isomers), Ani s 3 (2 isomers), Ani s 4, Ani s 8, Ani s 9, Ani s 10, Ani s 11-like, Ani s 13, Ani s fructose 1,6-bisphosphatase, Ani s phosphatidylethanolamine-binding protein (PEPB), and Thu a 3.0101. The following 8 allergens were detected in P. decipiens: Ani s 2, Ani s 3 (2 isomers), Ani s 5, Ani s 8, Ani s 9, Ani s PEPB, and Ani s troponin. In C. osculatum 4, the following allergens were identified: Ani s 2, Ani s 5, Ani s 13, and Asc l 3. Furthermore, 28 probable allergens were predicted in A. simplex and P. decipiens, whereas in C. osculatum, 25 possible allergens were identified. Among the putative allergens, heat shock proteins were most frequently detected, followed by paramyosin, peptidyl-prolyl cis-trans isomerase, enolase, and tropomyosin. We provide a new proteomic data set that could be beneficial for the discovery of biomarkers or drug target candidates. Furthermore, our findings showed that in addition to A. simplex, P. decipiens and C. osculatum should also be considered as potential sources of allergens that could lead to IgE-mediated hypersensitivity.

Morphological and molecular evaluation of tapeworms of the genus Bothriocephalus Rudolphi, 1808 (Cestoda: Bothriocephalidea), based on newly collected and uniformly fixed worms from freshwater fishes in Canada and the United States has revealed unexpected diversity. With a combination of selected morphological features and 4 molecular markers (18S rDNA V8 region, ITS1, ITS2, and COI gene sequences), the following morphotypes and lineages of the Bothriocephalus cuspidatus Cooper, 1917 complex were identified, several of which are specific to their respective fish definitive hosts and may represent separate species: B. cuspidatus sensu stricto from walleye, Sander vitreus (type host), which likely includes a miniature morphotype from Johnny darter, Etheostoma nigrum (both Percidae); Bothriocephalus morphotype from pumpkinseed, Lepomis gibbosus (Centrarchidae); and Bothriocephalus morphotype from rock bass, Ambloplites rupestris (Centrarchidae). The Bothriocephalus morphotype from goldeye, Hiodon alosoides (Hiodontidae), may also represent a separate lineage (possibly Bothriocephalus texomensisSelf, 1954) but requires additional studies. A morphotype from smallmouth bass, Micropterus dolomieu, based on a single specimen, is morphologically and genetically very similar to the morphotype from rock bass. Morphological study of the scolex and strobila of heat-killed and fixed specimens has revealed consistent differences, often subtle, that allowed us to differentiate between these morphotypes.

Trichuriasis is a serious threat to the economic development of animal husbandry. This research aimed to establish a droplet digital PCR (ddPCR) method to detect Trichuris spp. for the early diagnosis and prevention of trichuriasis in sheep. The real-time quantitative PCR (qPCR) and ddPCR methods were used for the detection of nematodes by targeted amplification of the ITS gene. Each means was evaluated to optimize the limit of detection and reproducibility. For a recombinant plasmid, the qPCR results showed that the detection limit was 31.7 copies per reaction. In contrast to qPCR, ddPCR was able to detect concentrations below 3.17 copies per reaction. Both assays exhibited good reproducibility. However, the ddPCR method was more stable for low-copy-number detection. This new assay was specific for Trichuris spp. and did not cross-react with other relevant gastrointestinal nematodes. A total of 98 clinical samples were tested with both assays. The results showed that the positive rate of ddPCR (80.6%) was higher than that of qPCR (72.4%). This method could be used as an efficient molecular biology tool to test for Trichuris spp. and could be a new valuable tool for the clinical diagnosis and prevention of trichuriasis.

Oxyuricassis ekstromi n. sp. is the third species of the genus described from the intestine of Lasiancistrus saetiger Armbruster in Brazil. This species can be easily differentiated from its congeners by the presence of a tooth-like sclerotized piece that occupies half of the stoma and absence of spine projections on the tail of both males and females. Also, males of the new species lack caudal alae present in Oxyuricassis coronatus and Oxyuricassis hexaspinatus. Additionally, we observed a different number and pattern of the distribution of cloacal papilla; males of O. coronatus and O. hexaspinatus have 1 pair of pre-cloacal and 2 pairs of post-cloacal papillae; while the new species has 1 pre-cloacal and 3 post-cloacal pairs of papillae.

Here we describe a new heterophyid species, Heterophyes yacyretana n. sp., and resolve its life cycle experimentally. We found the prosobranch snail Aylacostoma chloroticum in Candelaria, Province of Misiones, Argentina (a sector of the High Paraná River affected by the Yacyretá Dam), naturally infected with opisthorchioid cercariae. These cercariae lacked pigmented eyespots as well as body pigment and possessed 7 pairs of penetration glands arranged in 2 lateral bands, together with 18 pairs of flame cells and a V-shaped excretory vesicle. We exposed specimens of 21 fish species to emerging cercariae and obtained metacercariae from the muscles of the caudal peduncle of 3 species of siluriform fish, and adults from chicks infected with experimentally obtained metacercariae from the albino variety of the bronce corydoras, Corydoras aeneus. The new species differs from other species in the genus by the number of sclerites on the genital sac, the distribution of the vitelline follicles, and the combination of the size relationship of the suckers and the genital sac with respect to the posterior extent of intestinal ceca. Heterophyes yacyretana is the first species of the genus reported from the Americas.

Salamanders of the tribe Bolitoglossini Hallowell are a highly diversified group of amphibians, and their helminth parasite fauna has been scarcely studied. Some species of plethodontid salamanders distributed along the Trans-Mexican Volcanic Belt, in central Mexico, were sampled, and their helminth parasites were recovered for taxonomic identification. Specimens of a pharyngodonid nematode from 2 species of bolitoglossines of the genus Pseudoeurycea Taylor were morphologically identified as Batracholandros salamandrae (Schad, 1960) Petter and Quentin, 1976. These specimens were studied in further detail through light and scanning electron microscopy and were sequenced for 2 ribosomal genes and 1 mitochondrial gene to test the hypothesis of whether B. salamandrae is a species widely distributed in salamanders across the Nearctic biogeographic region, or if it represents a cryptic species complex. Our molecular results revealed that these specimens consisted of 2 genetic lineages in concordance with host species, although with slight morphological differences among specimens in each of them. A thorough study, including the generation of molecular data from individuals from other areas of North America, and the examination of type specimens, is required to test the reliability of these morphological differences and to corroborate the species identity of the 2 genetic lineages.

Between June 2016 and June 2019, we surveyed 62 Mediterranean geckos, Hemidactylus turcicus, from Abu Rawash, Giza, Egypt, for the presence of endoparasites. In June 2016, we found 3 individuals to be infected with Eimeria lineri. We studied the morphology and inner structures of its sporulated oocysts, and the locations of its intestinal endogenous stages. We also extracted genomic DNA from these sporulated oocysts and successfully sequenced a 632-bp fragment of the 18S rRNA gene. Phylogenetic analyses using this partial sequence allowed us to support previous studies that assigned E. lineri to the genus Acroeimeria. Our consensus sequence was used to query similar 18S rDNA sequences from GenBank, and 14 sequences were selected. The phylogenetic analysis inferred by maximum likelihood and Bayesian inference methods gave similar results, as both separated the sequences into 2 clades: (1) a monophyletic group of Goussia species (from fish); and (2) a strongly supported clade that separated 4 Choleoeimeria species from a polyphyletic group of species that clustered A. lineri with 3 other Acroeimeria species and 3 Eimeria species from lizards, including Eimeria tiliquae from Tiliqua rugosa (Gray, 1825), Eimeria tokayae from Gecko gecko (L., 1758), and Eimeria eutropidis from Eutropis macularia (Blyth, 1853). Our study supports the placement of E. lineri into the Acroeimeria and contributes additional life history information toward understanding the evolutionary origin of the Eimeria-like species that have sporocysts without Stieda bodies in their oocysts and that infect saurian reptiles. We also support the concept that several traits (morphological, endogenous, and gene sequences) are both necessary and important for authors to include when making generic reassignments within the eimeriid coccidia.

We determined the complete sequence of the mitochondrial DNA (mtDNA) of a parasite discovered between the subcutaneous tissue and the peritoneum of an African nocturnal non-human primate (NHP). The parasite and host sequences were obtained by a combination of Sanger sequencing and nanopore MinION techniques. Analyses of mtDNA gene arrangements and sequences unambiguously showed that the parasite investigated was the pentastomid Armillifer armillatus, also commonly named the tongue worm. The full-length mitochondrial genome of A. armillatus, measuring 16,706 bp in length, contains 13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes, an arrangement identical to that of previously described pentastomid mitochondrial genomes. We describe here the second full mitochondrial genome of A. armillatus to date. To identify the NHP host, maximum likelihood phylogenetic analyses of a 441-bp fragment on the 12S rDNA gene and of a 1,140-bp fragment of the mitochondrial cytochrome b strongly support clustering with the African lorisid Perodicticus potto, a species that has rarely been reported as an intermediate host of this parasite.

Ectoparasites were collected from Eptesicus hottentotus, the long-tailed serotine bat, caught in Namibia as part of an ecological study. Larvae of Argas transgariepinus, a blood-feeding ectoparasite of bats in Africa, were removed from 3 of 18 bats. We present scanning electron microscope images of unengorged larvae. As with other ectoparasites, this bat tick might transmit pathogens such as Borrelia and Rickettsia to their hosts as has been reported for bat ticks in Europe and North America. We screened 3 pools (25 total) of larvae of A. transgariepinus removed from the long-tailed serotine bat Eptesicus hottentotus caught in Namibia. Two microbes of unknown pathogenicity, including Rickettsia hoogstraalii, a spotted fever group pathogen, and a Rickettsiella sp. were detected by molecular techniques.

Introduction of exotic tick vectors of bacteria, protozoa, viruses, and filarial parasites into the United States has accelerated in recent years, primarily because of globalization, increased frequency of travel, and a rise in legal and illegal animal trades. We herein report introduction of a live specimen of Amblyomma oblongoguttatum on a human into the United States from Central America, and we review 4 previous similar incidents. This tick species occurs widely in the neotropics, from western and southern Mexico, southwards through Central America, to the northern half of South America. It is a potential vector of bacterial agents of spotted fever group rickettsioses, raising concern that if A. oblongoguttatum ticks become established in this country, they might also be able to carry pathogens of human and veterinary concern. Given the potential for exotic ticks as vectors of numerous pathogens, proper surveillance, interception, and identification of these ticks are vital to protecting human and veterinary health. Rigorous governmental inspections of imported livestock and pet animals at ports of entry and educating human travelers and medical practitioners about the risks should be part of an overall national tick program.

Bess beetles (Passalidae) display important roles in forestall ecosystems, particularly in energy extraction from dead wood. These organisms maintain complex biological interactions with their gut symbiotic communities, including bacteria, protists, and metazoans. Very little is known about symbionts since most of the species of Passalidae haven't been studied from a parasitological point of view. Here we describe a new genus and 2 new species of nematodes of the family Hystrignathidae associated with 2 beetle species of the tribe Proculini collected in the State of Oaxaca, Mexico. Tuhmai garciaprietoi n. gen., n. sp., found in Vindex agnoscendus is characterized by the presence of an unarmed cervical cuticle, a subcylindrical procorpus and a conspicuous isthmus, a monodelphic-prodelphic reproductive system, and a short subulate tail. Urbanonema osorioi n. sp., found in Verres hageni mainly differs from other species of Urbanonema by the number and disposition of cervical spines, as well as by a subulate tail. For each new taxon, we describe the external and internal morphology, and we generated molecular data (nuclear ribosomal DNA) to place the new taxa in a phylogenetic context.

Information regarding trichodinid ectoparasites on marine fishes of North America is relatively scarce. In this study, 5 ciliate species from the family Trichodinidae were found associated with 8 fish hosts from the rocky intertidal zone of the western coast of the Baja California Peninsula (BCP), Mexico. All of the host-parasite relationships recorded here are new. Furthermore, 3 of the trichodinid species found are recorded for the first time for Mexico. Trichodinids taxa do not show a noticeable distributional gradient along the BCP, which suggests a wide-continuous distribution of the species throughout the study area.

Giardia duodenalis is a common zoonotic protozoan parasite with a broad host distribution. The main objectives of the present study were to determine the prevalence of giardiasis and to reveal the genetic and haplotype diversity of G. duodenalis in symptomatic cats in Turkey. Fecal samples were collected from cats (n = 102) with diarrhea that were admitted to different pet clinics in the Central Anatolia region of Turkey. All samples were analyzed by microscopic examination (ME), rapid immunochromatographic test (ICT), and PCR targeting the β-giardin (bg) loci of the parasite. Phylogenetic, haplotype, and network analyses of G. duodenalis based on the bg gene were carried out. Overall, G. duodenalis was detected in 70/102 (68.6%) of the cats with diarrhea by ME (38/102, 37.3%), ICT (51/102, 50%), and PCR (30/102, 29.4%). According to sequence analyses of the bg gene region, all isolates were identified as G. duodenalis assemblage B. Haplotype analyses revealed 2 known and 8 novel haplotypes for G. duodenalis assemblage B. This study provides first prevalence and genetic and haplotype diversity data on G. duodenalis assemblage B from cats in Turkey.

Water mites of the genus Unionicola are common parasites of freshwater mussels, living on the gills or mantle of their hosts and using these tissues as sites of oviposition. Although surveys of this mite fauna among North American unionid mussels indicate that these mites represent highly diverse assemblages, we know very little regarding the determinants of Unionicola species diversity among their molluscan hosts. The present study addresses the relationship between host diversity and mite diversity for Unionicola assemblages associated with unionid mussels of North America. The results of this study found a significantly positive relationship between host species richness and mite species richness, adding to a growing body of evidence that host diversity is an important determinant of parasite diversity. In recent years, molecular sequence data have discovered cryptic biodiversity among unionid mussels, yielding revisions in the nomenclature and systematic taxonomy of the group. DNA sequence variation has also revealed cryptic species complexes among Unionicola mites. Collectively, these findings suggest that the results of the present study may be underestimating species richness among mites and their host mussels. Unfortunately, human perturbations are known to have caused high recent rates of extinction in the mussel and mite faunas of North America and could play a major role in influencing patterns of species richness for this host–parasite system moving forward.