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Morphological characteristics of the acanthocephalan Polymorphus minutus (Goeze, 1782), which was collected from the duck Anas platyrhynchos Linnaeus in the Czech Republic, are described. The mitochondrial (mt) genome of P. minutus was sequenced, with a total length of 14,149 bp, comprising 36 genes including 12 protein coding genes (PCGs), 22 transfer RNA (tRNA) genes and two ribosomal RNA genes (rrnL and rrnS). This genome is similar to the mt genomes of other syndermatan species. All these genes were encoded on the same DNA strand and in the same orientation. The overall nucleotide composition of the P. minutus mt genome was 38.2% T, 27.3% G, 26.2% A, and 8.3% C. The amino acid sequences of 12 PCGs for mt genomes of 28 platyzoans, including P. minutus, were used for phylogenetic analysis, and the resulting topology recovers P. minutus as sister to Southwellina hispida (Van Cleave, 1925), and the two taxa form a sister clade to Centrorhynchus aluconis (Müller, 1780) and Plagiorhynchus transversus (Rudolphi, 1819), which are all species in the Palaeacanthocephala, thus supporting the monophyly of this class.

Tapeworms of the genus Proteocephalus Weinland, 1858 (Cestoda: Proteocephalidae) are common and widespread intestinal parasites of whitefish (Coregonus spp., Salmonidae: Coregoninae). Previous taxonomic studies, based solely on morphology and inconsistently fixed specimens, concluded that all salmoniform fish, including whitefish, are parasitised by a single euryxenous and highly polymorphic species, Proteocephalus longicollis (Zeder, 1800). However, recent molecular phylogenetic analyses have revealed the existence of several species specific to individual genera or even species of salmoniform fish. In this study, Proteocephalus fallax La Rue, 1911 is redescribed based on newly collected and genetically characterised specimens from several Coregonus species in Switzerland, the type locality of the species, and in Norway. This cestode was previously synonymised with P. exiguus La Rue, 1911, a parasite of whitefish in North America, but the two species are not closely related. Proteocephalus fallax differs from P. exiguus in its larger body size, wider proglottids, shorter cirrus sac and broader scolex. In addition, the other Proteocephalus species described in whitefish are briefly discussed, with comments on their validity, host range and distribution.

The nematode Spinitectus notopteri Karve et Naik, 1951 was collected from two species of freshwater fish belonging to the family Notopteridae, namely Chitala ornata (Gray) and Notopterus notopterus (Pallas), at Khun Thale Swamp in Surat Thani province of southern Thailand. The overall prevalence of the parasite was found to be 88% (94 fish infected/106 fish examined). A higher prevalence was found in C. ornata (96%), while a higher intensity was noted for N. notopterus (117 parasites/fish). The most important morphological characters were the presence of cephalic and cuticular structures, precloacal ridges, and genital papillae. Specimens were genetically characterised using cox1 mtDNA. Morphological characteristics were most similar to Spinitectus petterae Boomker, 1993 from Clarias gariepinus (Burchell) in Africa, while the genetic data were dissimilar to all available data for the genus. Therefore, the identification of nematode specimens using a combination of morphological and molecular techniques is stressed. This study also presents the first molecular analysis of S. notopteri infecting Thai freshwater fish and a new geographical record for S. notopteri.

Chagas disease (CD) is a neglected disease caused by Trypanosoma cruzi Chagas, 1909. Causative treatment can be achieved with two drugs: benznidazole or Nifurtimox. There are some gaps that hinder progress in eradicating the disease. There is no test that can efficiently assess cure control after treatment. Currently, the decline in anti-T. cruzi antibody titres is assessed with conventional serological tests, which can take years. However, the search for new markers of cure must continue to fill this gap. The present study aimed to evaluate the decline in serological titres using chimeric proteins after treatment with benznidazole in chronic patients diagnosed with CD. It was a prospective cross-sectional cohort study between 2000 and 2004 of T. cruzi-positive participants from the Añatuya region (Argentina) treated with benznidazole. Serum samples from ten patients were collected before treatment (day zero) and after the end of treatment (2, 3, 6, 12, 24 and 36 months). For the detection of anti-T. cruzi antibodies, an indirect ELISA was performed using two chimeric recombinant proteins (IBMP-8.1 and IBMP-8.4) as antigens. The changes in reactivity index within the groups before and after treatment were evaluated using the Friedman test. All participants experienced a decrease in serological titres after treatment with benznidazole, especially IBMP-8.1. However, due to the small number of samples and the short follow-up period, it is premature to conclude that this molecule serves as a criterion for sustained cure. Further studies are needed to validate tests based on these or other biomarkers to demonstrate parasitological cure.

Morphological data are used to describe a new nematode species, Heterocheilus floridensis sp. n. (Heterocheilidae) from the digestive tract of the Florida manatee Trichechus manatus latirostris (Harlan) (Trichechidae, Sirenia) from Florida, USA. Examination by light and scanning electron microscopy revealed that the new species differs from the related Heterocheilus tunicatus Diesing, 1839 mainly by having dentigerous ridges on the inner surface of the lips, a median unpaired papilla located anterior to the cloaca, and a considerably larger body size. Sequence data for subunits I and II of mitochondrial cytochrome oxidase gene, 18S small subunit and 28S ribosomal RNA genes were provided for molecular characterisation of the new species. However, the current unavailability of homologous sequence data for congeneric specimens precluded a molecular assessment of the morphological species hypothesis, and ascaridoid phylogenetic hypotheses could not be advanced. Specimens of Heterocheilus sp. collected from the Antillean manatee Trichechus manatus manatus Linnaeus in Puerto Rico, on loan from the US National Museum of Natural History, were morphologically consistent with the new species, so apparently all congeneric nematodes reported from both subspecies of the West Indian manatee Trichechus manatus Linnaeus and previously identified as H. tunicatus belong rather to H. floridensis sp. n. Heterocheilus hagenbecki (Khalil et Vogelsang, 1932) Sprent 1980 is here considered to be a species inquirenda. A key to valid species of Heterocheilus Diesing, 1839 is provided.

The present paper comprises a systematic survey of helminths (trematodes, an acanthocephalan and nematodes) found in nine species of freshwater fishes in Ecuador collected in March 1999 and those (a trematode and acanthocephalans) collected from an amphibian and two species of freshwater fishes in Venezuela in 1992, 1996 and 2001. The following 17 helminth species were recorded: Trematoda: Prosthenhystera ornamentosa sp. n., P. obesa (Diesing, 1850), Crassicutis intermedius (Szidat, 1954), C. cichlasomae Manter, 1936 and Glypthelmins eleutherodactyli sp. n. Acanthocephala: Quadrigyrus torquatus Van Cleave, 1920, Gracilisentis variabilis (Diesing, 1851) and Neoechinorhynchus (Neoechinorhynchus) ecuadoris sp. n. Nematoda: Cosmoxynema vianai Travassos, 1949, Travnema travnema Pereira, 1938, Touzeta ecuadoris Petter, 1987, Sprentascaris hypostomi Petter et Cassone, 1984, Sprentascaris sp., Contracaecum sp. Type 1 larvae, Contracaecum sp. Type 2 larvae, Procamallanus (Procamallanus) peraccuratus Pinto, Noronha et Rolas, 1976 and Procamallanus (Spirocamallanus) sp. juv. Nearly all of these parasites are reported from Ecuador or Venezuela for the first time and many of these findings represent new host records. The new species P. ornamentosa sp. n. was collected from the gall-bladder of an unidentified anostomid (Anostomidae, Characiformes) in Ecuador, G. eleutherodactyli sp. n. from the digestive tract of the frog Eleutherodactylus sp. (Eleutherodactylidae, Anura) in Venezuela and N. (N.) ecuadoris sp. n. from the intestine of Lebiasina sp. (Lebiasinidae, Characiformes) in Ecuador. Most parasites are briefly described and illustrated and problems concerning their morphology, taxonomy, hosts and geographical distribution are discussed.

Liver pathologies and infection with Toxoplasma gondii (Nicolle et Manceaux, 1908) are widespread among HIV-infected patients. However, a possible contribution of toxoplasmosis to the development of various forms of liver diseases in HIV-infected individuals has not yet been determined. This research is a retrospective cohort study. Medical cards of 907 HIV-positive patients, including 119 individuals who died, were studied. The patients were divided into two groups: 531 patients were seropositive to T. gondii and 376 seronegative. General liver pathology was more widespread among patients seropositive to T. gondii than in seronegative patients (63.1 ± 2.1% and 51.9 ± 2.6%, respectively, p < 0.001). The association of seropositive to T. gondii with general liver pathology is weak both in the whole cohort (Pearson's contingency coefficient C = 0.112), and among the deceased patients (C = 0.228). Chronic HBV-HCV coinfection was more common in the seropositive than in seronegative individuals as it was found both in entire cohorts (26.0 ± 1.9% and 18.6 ± 2.0%, respectively, p = 0.010) and in died patients (31.0 ± 5.5% and 14.6 ± 5.1%, respectively, p = 0.041). Toxoplasma gondii had a weak role in distributing of HBV-HCV coinfection between cohorts (C = 0.187). In both cohorts in patients with chronic hepatitis, regardless of its etiology, there was no significant difference in alanine transaminase activity (ALT). Cirrhosis of the liver occurred 4.5 times more often in deceased seropositive patients than in the entire seropositive cohort (23.9 ± 5.1 and 5.3 ± 2.0, respectively, p = 0.0006) whereas it no significantly increased in seronegative cohort (10.4 ± 4.4 against 4.8 ± 1.1, p > 0.05). In them T. gondii is weakly involved in cirrhosis formation (C = 0.168). Thus, in HIV-infected patients, T. gondii is a weak nonspecific adjunct that supports chronic liver inflammation and progression of cirrhosis, regardless of etiology, but does not influence the degree of hepatitis activity. The increased prevalence of HBV-HCV coinfection in patients seropositive for T. gondii may be related to their risk factor behaviour associated with uncontrolled blood contacts.

Hexokinase (HXK) is the first key enzyme in the glycolytic pathway and plays an extremely important role in energy metabolism. By searching the microsporidian database, we found a sequence (NBO_27g0008) of Nosema bombycis Nägali, 1857 with high similarity to hexokinase-2, and named it as NbHXK2. The NbHXK2 gene has 894 bp and encodes 297 amino acids with 34.241 kD molecular weight and 5.26 isoelectric point. NbHXK2 contains 31 phosphorylation sites and 4 potential N-glycosylation sites with signal peptides and no transmembrane domain. Multiple sequence alignment showed that NbHXK2 shares more than 40% amino acid identity with that of other microsporidia, and the homology with hexokinase-2 of Nosema tyriae Canning, Curry, Cheney, Lafranchi-Tristem, Kawakami, Hatakeyama, Iwano et Ishihara, 1999, Nosema pyrausta (Paillot, 1927) and Nosema ceranae Fries, Feng, da Silva, Slemenda et Pieniazek, 1996 was 89.17%, 87.82% and 69.86%, respectively. Phylogenetic analysis based on the amino acid sequence of hexokinase showed that all microsporidia cluster together in the same clade, and are far away from animals, plants and fungi, and that N. bombycis is closely related to N. tyriae; N. pyrausta; N. ceranae and Nosema apis Zander, 1909. Immunolocalisation with the prepared polyclonal antibody showed that NbHXK2 was mainly distributed in the cytoplasm and plasmalemma in proliferative, sporulation stage and mature spore of N. bombycis. qRT-PCR assay showed that the NbHXK2 expressed at higher level during spore germination and at early stage of proliferation. These results indicate that N. bombycis may use its own glycolytic pathways to supply energy for infection and development, especially germination and in the early stage of proliferation, and acquire energy from the host through certain ways as well.

Recent examinations of some marine fishes from off the southern coast of Iraq revealed the presence of five species of Philometra Costa, 1845 (Nematoda: Philometridae): Philometra arabiensis sp. n. (males and females) from the ovary of the shrimp scad Alepes djedaba (Forsskål) (Carangidae, Carangiformes), Philometra psettoditis Moravec, Walter et Yuniar, 2012 (females) from the body cavity of the Indian halibut Psettodes erumei (Bloch et Schneider) (Psettodidae, Pleuronectiformes), Philometra terapontis Moravec, Gopalakrishnan, Rajkumar, Saravanakumar et Kaliyamoorthy, 2011 (female) from the ovary of the jarbua terapon Terapon jarbua (Forsskål) (Terapontidae, Centrarchiformes), Philometra sp. (females) from the ovary of the Arabian blackspot threadfin Polydactylus mullani (Hora) (Polynemidae, Carangariformes) and Philometra sp. 2 of Moravec et al. (2016a) (females) from the ovary and body cavity of the bartail flathead Platycephalus indicus (Linnaeus) (Platycephalidae, Perciformes). Philometra arabiensis sp. n. is mainly characterised by the length of spicules (198-243 µm) and gubernaculum (75-99 µm), the gubernaculum/spicule length ratio (1 : 2.33-2.79), the structure of the gubernaculum distal portion and the male caudal end, and the body length of males (1.86-2.73 mm). The present findings of P. psettoditis and P. terapontis in fishes of the Arabian Gulf represent new geographical records for these parasites.

Examinations of nematodes collected from some marine fishes off the southwestern coast of Java, Indonesia in 2000 and 2001 revealed the presence of the following six species: ascaridoids Ichthyascaris grandis sp. n. from the intestine of Lophiomus setigerus (Vahl), I. cf. longispicula Li, Liu, Liu et Zhang, 2012 from the intestine of Conger cinereus Rüppel, Ichthyascaris sp. from the body cavity of Lobotes surinamensis (Bloch), and Raphidascaroides halieutaeae Yin, 1983 from the intestine of Halieutaea stellata (Vahl), and philometrids Philometra ivaschkini Parukhin, 1976 from the stomach wall of Trichiurus lepturus Linnaeus and P. psettoditis Moravec, Walter et Yuniar, 2012 from the body cavity (liver) of Psettodes erumei (Bloch et Schneider). Descriptions of these nematodes based on light and scanning electron microscopical studies are provided. The new species I. grandis sp. n. is mainly characterised by large body measurements (males and females up to 41.8 mm and 73.6 mm long, respectively), the length of spicules (0.99-1.05 mm), the tail tip usually without rudimentary spines and by the presence of 44-53 pairs of caudal papillae, eight to twelve of which being postanals. In addition to new data on the morphology of R. halieutaeae and other nematodes recorded, the 11 species of Raphidascaroides Yamaguti, 1941 poorly described from marine fishes in South Asia and reviewed in the monograph of Sood (2017) are considered species inquirendae and incertae sedis.

Adult specimens of five little-known species of spiruridan nematodes (Spirurida) were collected during occasional examinations of some fishes from the Poonch River in Jammu and Kashmir, India: Procamallanus (Spirocamallanus) bilaspurensis Gupta et Duggal, 1973 in Mastacembelus armatus (Lacépède) (Mastacembelidae, Synbranchiformes), Rhabdochona (Rhabdochona) indica Moravec, Scholz, Ash et Kar, 2010 in Crossocheilus latius (Hamilton) (Cyprinidae, Cypriniformes), Rhabdochona (Rhabdochona) turkestanica (Skryabin, 1917) in Schizothorax richardsonii (Gray) (Cyprinidae, Cypriniformes) and both Rhabdochona (Rhabdochona) hospeti Thapar, 1950 and Rhabdochona (Globochona) cf. chodukini Osmanov, 1957 in Tor putitora (Hamilton) (Cyprinidae, Cypriniformes); besides these, unidentifiable physalopterid larvae were recorded from M. armatus and T. putitora. Their detailed light and electron microscopical studies revealed some taxonomically important, previously not observed features and made possible their redescription. Fourth-stage larvae of P. (S.) bilaspurensis and R. (R.) turkestanica are described for the first time. The latter is resurrected as an independent species. The following are considered to be species inquirendae: a total of nine poorly described species of Rhabdochona Railliet, 1916 from T. putitora in Balochistan, Pakistan, all listed in the monograph of Kakar et al. (2011), plus an additional 15 species of the genus reported from cyprinids and a catfish in the same region (Balochistan) by the same authors. Also three congeneric species from India (R. nemacheili Rautela et Malhotra, 1982, R. putitora Kaur et Khera, 1991 and R. tori Gupta et Srivastava, 1982) and one from Pakistan (R. charsaddiensis Siddiqi et Khattak, 1984) should be considered as species inquirendae due to their poor descriptions. The names of Rhabdochona putitori Anjum, 2013, R. indusi Soofi, Birmani et Dharejo, 2017 and R. sindhicus Soofi, Birmani, Dharejo, Abbasi et Ghachal, 2020 are invalid according to the International Code of Zoological Nomenclature.

Evolutionary and ecological processes affecting the interactions between hosts and parasites in the aquatic environment are at display in the Baltic Sea, a young and ecologically unstable marine ecosystem, where fluctuating abiotic and biotic factors affect the parasitofauna in fish. The dynamic infections of Baltic cod, a subpopulation of the Atlantic cod (Gadus morhua Linnaeus), with third stage anisakid nematode larvae of Pseudoterranova decipiens (Krabbe, 1878) and Contracaecum osculatum (Rudolphi, 1802) have increased following a significant increase of the Baltic grey seal Halichoerus grypus (Fabricius) population in the region. Cod serves as a paratenic host and marine mammals, pinnipeds, are definitive hosts releasing parasite eggs, with faeces, to the marine environment, where embryonation and hatching of the third stage larva take place. The parasite has no obligate intermediate hosts, but various invertebrates, smaller fish and cod act as paratenic hosts transmitting the infection to the seal. Contracaecum osculatum has an impact on the physiological performance of the cod, which optimises transmission of the larva from fish to seal. Thus, a muscle mass decrease of nearly 50% may result from heavy C. osculatum infections, probably amplified by a restricted food availability. The muscle atrophy is likely to reduce the escape reactions of the fish when meeting a foraging seal. In certain regions, where fish and seals are restricted in their migration patterns, such as the semi-enclosed Baltic Sea, the predation may contribute to a severe cod stock depletion. The parasites are zoonotic and represent a human health risk, when consumers ingest insufficiently heat- or freeze-treated infected products. Marked infections of the cod were previously reported during periods with elevated seal populations (late 19th and middle 20th century) and various scenarios for management of risk factors are evaluated in an evolutionary context.

Two species of Spinitectus Fourment, 1884 have been recorded from southern Africa, namely Spinitectus polli Campana-Rouget, 1961 and Spinitectus petterae Boomker, 1993, both from the Limpopo River system. Spinitectus petterae was described from North African catfish, Clarias gariepinus (Burchell), whereas S. polli infects squeakers, Synodontis spp. During parasitological surveys in the Vaal River system (Orange River catchment), Spinitectus specimens were collected from C. gariepinus. These systems are adjacent but not connected. Therefore, this study aimed to identify the specimens collected using morphological and molecular techniques. The morphological study included light and scanning electron microscopy of whole specimens and excised spicules. Specimens were genetically characterised using 18S rDNA, 28S rDNA and cox1 mtDNA. Additionally, immature specimens of S. petterae were collected near the type locality. Morphological characteristics were most similar to S. petterae from C. gariepinus, whereas genetic data were dissimilar to all available data for the genus. Additional morphological characteristics noted for S. petterae in the present study were the details of the left and right spicule structure and the porous structures on the pseudolabia. Specimens from the Vaal River system differed from those originally described as S. petterae by additional spines posterior to the third ring, lacking caudal alae and variable total body and male oesophagus length. Based on 18S rDNA, haplotypes from the type locality varied only slightly from the study material, supporting the morphological identification. However, 28S rDNA and, more conspicuously, cox1 mtDNA displayed substantial variation between specimens from these localities, which needs further investigation. Haplotypes generated in the present study were highly dissimilar to those characterised for S. petterae from Tanzania and Egypt. Nevertheless, the nematodes collected from C. gariepinus in the Vaal River system are considered S. petterae. This study expands the geographical distribution and adds additional morphological and genetic information for S. petterae, contributing to the limited knowledge of African species of Spinitectus.

Eudiplozoon nipponicum (Goto, 1891) Khotenovsky, 1985 (Monogenea: Diplozoidae), is known to parasitise Cyprinus carpio Linnaeus and species of Carassius. In this study, we conducted a taxonomic re-examination of E. nipponicum using genetic analysis and morphological comparisons from different host species from a single water system. rDNA nucleotide sequences of the internal transcription spacer 2 (ITS-2) region (645 bp) showed interspecific-level genetic differences among diplozoids from species of Carassius and C. carpio (p-distance: 3.1-4.0%) but no difference among those from different species of Carassius (0-0.4%) or between those from C. carpio collected in Asia and Europe (0-1.1%). Large variation was observed among 346 bp cytochrome c oxidase subunit I (COI) sequences (0.3-16.0 %); the topology of the phylogenetic tree showed no relationship to host genera or geographical regions of origin. Morphological observation showed that average clamp size of diplozoids from C. carpio was larger than those from Carassius spp. The number of folds on the hindbody was 10-25 for diplozoids from C. carpio and 12-19 for those from Carassius spp. Thus, our ITS-2 sequence and morphological comparison results indicate that diplozoids from C. carpio and species of Carassius belong to different species. The scientific name E. nipponicum should be applied to the species infected to the type host, Carassius sp. of Nakabo (2013) (Japanese name ginbuna). The diplozoid infecting C. carpio (Eurasian type) should be established as a new species: Eudiplozoon kamegaii sp. n. A neotype of E. nipponicum is designated in this report because the original E. nipponicum specimens are thought to have been lost.

We provide the first ultrastructural evidence of the secretion of extracellular vesicles (EVs) across all parasitic stages of the tapeworm Schistocephalus solidus (Müller, 1776) (Cestoda: Diphyllobothriidea) using a laboratory life cycle model. We confirmed the presence of EV-like bodies in all stages examined, including the hexacanth, procercoids in the copepod, Macrocyclops albidus (Jurine, 1820), plerocercoids from the body cavity of the three-spined stickleback, Gasterosteus aculeatus Linnaeus, and adults cultivated in artificial medium. In addition, we provide description of novel tegumental structures potentially involved in EV biogenesis and the presence of unique elongated EVs similar to those previously described only in Fasciola hepatica Linnaeus, 1758 (Trematoda), Hymenolepis diminuta (Rudolphi, 1819) (Cestoda), and Trypanosoma brucei Plimmer et Bradford, 1899 (Kinetoplastida).

Toxoplasmosis is one of the world's most prevalent zoonoses. The causative agent, Toxoplasma gondii (Nicolle et Manceaux, 1908) is a facultative heteroxenic, polyxenic apicomplexan protist. There are several potential pathways of transmission within and between host species. Most infections with T. gondii result from close contact with pets/cats, ingestion of tissue cysts in undercooked meat of infected animals, and oocysts from food or water contaminated by feline faeces. Recently, epidemiological studies have shown that T. gondii infection plays a prominent role in the pathogenesis of several psychiatric disorders. This report reviews the association between T. gondii infection and patients with psychiatric disorders, particularly schizophrenia, depressive disorders and bipolar disorders.

Females of species of Lamproglena von Nordmann, 1832 are parasitic on the gills of teleost fishes and the 38 nominal species are based on mainly morphological data. Only four of these species have been genetically characterised and no mitochondrial data are available for the genus. The present study aimed to provide representative ribosomal DNA (rDNA) data for two additional species of Lamproglena from Africa: Lamproglena clariae Fryer, 1956 and Lamproglena hoi Dippenaar, Luus-Powell et Roux, 2001, alongside mitochondrial DNA (mtDNA) for these and two other African species, Lamproglena hemprichii von Nordmann, 1832 and Lamproglena monodi Capart, 1944. The four species were collected from Clariidae, Cyprinidae, Alestidae and Cichlidae, respectively. Representative 18S rDNA and 28S rDNA data were obtained for L. clariae and L. hoi, while cox1 mtDNA was obtained for all four species. The respective haplotypes supported the distinctness of all species using all three gene regions investigated. Interestingly, species appeared to be grouped more by geographical origin than host family, with L. hoi more closely related to other African species than to Asian species also collected from cyprinid hosts. Even though the results presented here greatly add to the molecular data available for Lamproglena, there are still 32 (>80%) species for which no genetic data are available. The interpretation of the results presented here is thus preliminary and much more data are required before the phylogeny of this genus, and other members of the family, such as Lernaea Linnaeus, 1758, can be studied appropriately.

The genus Coitocaecum Nicoll, 1915 is part of the most speciose digenean family, the Opecoelidae Ozaki, 1925, which is found globally in both freshwater and marine fishes. Fifteen opecoelid species have been reported from marine fishes in South Africa, yet only one species of Coitocaecum has been described from this region: Coitocaecum capense Bray, 1987. During an explorative study of the digeneans of the endemic, intertidal fish Clinus superciliosus (Linnaeus) from the Saldanha Bay area, Cape Town harbour, Hermanus, the Tsitsikamma section of the Garden Route National Park and Chintsa East in South Africa, a total of three distinct species of Coitocaecum were identified based on morphological and molecular (28S rDNA, ITS1-5.8S-ITS2 rDNA and COI mtDNA) data: the previously mentioned C. capense, Coitocaecum brayi sp. n. and a third, unnamed species. We provide the first molecular characterisation of species of Coitocaecum from South Africa, accompanied by detailed morphological descriptions. This study illustrates the importance of an integrated taxonomic approach, especially when studying species with similar morphology. These findings further emphasise the lack of information on the true diversity and molecular data for trematodes of marine fishes in South Africa, creating a great capacity for future explorative taxonomic studies and highlighting the use of intertidal areas for conducting such research.

As presently defined, the tapeworm genus Andrya Railliet, 1895 (Cyclophyllidea: Anoplocephalidae sensu stricto) includes the type species A. rhopalocephala (Riehm, 1881) in hares of the genus Lepus Linnaeus (Leporidae) in western Eurasia and four species in cricetid (Neotominae, Sigmodontinae) and octodontid rodents in North and South America. The host range of Andrya is puzzling, because it is the only genus of anoplocephalid (s. s.) cestodes parasitising both rodents and lagomorphs. The present morphological analysis shows that the American species of Andrya share multiple consistent features, in which they differ from those of A. rhopalocephala and the morphologically related Neandrya cuniculi (Blanchard, 1891). The main differences concern the position of the uterus with respect to the longitudinal osmoregulatory canals and testes. Consequently, a new genus Andryoides gen. n. is proposed for the American species, resulting in the following combinations: Andryoides neotomae (Voge, 1946) comb. n. (type species), Andryoides octodonensis (Babero et Cattan, 1975) comb. n., Andryoides vesicula (Haverkost et Gardner, 2010) comb. n. and Andryoides boliviensis (Haverkost et Gardner, 2010) comb. n. However, A. boliviensis is regarded here as a junior synonym of A. vesicula (new synonymy). The present study also defines the morphological key features for all the valid genera of cestodes of the family Anoplocephalidae (s. s.), and discusses the phylogenetic affinities and historical biogeography of Andryoides and other endemic American anoplocephalid cestodes.

A synopsis of 43 nominal species from five genera of tailed Myxobolidae infecting Indian freshwater and marine fishes is presented. The main characteristic of this group is the presence of at least one tail-like caudal process. For each species, relevant morphological and morphometric data are provided, such as the host(s), site(s) of infection within the host and sampling state. A key for the identification of 13 genera of tailed Myxobolidae is also included.

In an effort to expand knowledge of Clade 3-one of the ten clades that compose the non-monophyletic order 'Tetraphyllidea' all current members of which parasitise orectolobiform sharks-we targeted species of orectolobiform sharks that had not previously been examined for 'tetraphyllidean' cestodes. That work led to the discovery of three new species off Australia and Taiwan. Ambitalveolus gen. n. was erected to accommodate these species. Ambitalveolus costelloae gen. n. et sp. n., Ambitalveolus kempi sp. n., and Ambitalveolus penghuensis sp. n. differ from one another in scolex size, genital pore position, and number of marginal loculi, proglottids, and testes. Among 'tetraphyllideans', the new genus most closely resembles the two other genera in Clade 3. It differs from Carpobothrium Shipley et Hornell, 1906 in lacking anterior and posterior flap-like extensions of its bothridia; instead, its bothridia are essentially circular. It differs from Caulopatera Cutmore, Bennett et Cribb, 2010 in that its vitelline follicles are in two lateral bands, rather than circum-medullary, and in that its bothridia bear, rather than lack, conspicuous marginal loculi. A key to the three genera in Clade 3 is provided. A phylogenetic analysis including new sequence data for one of the three new species of Ambitalveolus gen. n., the only species of Caulopatera, and all four described species and one undescribed species of Carpobothrium supports previously hypothesised close affinities between Caulopatera and Carpobothrium, with the new genus as their sister group. This is the first report of 'tetraphyllidean' cestodes from the orectolobiform shark family Brachaeluridae Applegate. The association of the new species with orectolobiform sharks is consistent with those of the other members of Clade 3. However, whereas species of Carpobothrium and Caulopatera parasitise members of the hemiscylliid genus Chiloscyllium Müller et Henle, species of Amitalveolus gen. n. parasitise members of the Brachaeluridae and Orectolobidae Gill.

During an ichthyoparasitological survey in 2017-2019, six species of acanthocephalans were found among Taiwan's freshwater (Cypriniformes: Xenocyprididae, Cyprinidae) and marine fishes (Scombriformes: Scombridae, Trichiuridae; Anabantiformes: Channidae; Carangaria/misc: Latidae): Micracanthorhynchina dakusuiensis (Harada, 1938), Rhadinorhynchus laterospinosus Amin, Heckmann et Ha, 2011, Pallisentis rexus Wongkham et Whitfield, 1999, Longicollum sp., Bolbosoma vasculosum (Rudolphi, 1819), and one new species, Micracanthorynchina brevelemniscus sp. n. All species are morphologically characterised and illustrated using light and scanning electron microscopy. The finding of R. laterospinosus, P. rexus and B. vasculosum is the first record for these species in Taiwan. Micracanthorhynchina brevelemniscus is similar to Micracanthorhynchina motomurai (Harada, 1935) and M. dakusuiensis in proboscis armature but differs from M. motomurai by larger eggs (53-59 × 15-16 µm vs 40 × 16 µm) and by the number of cement glands (6 vs 4) and from M. dakusuiensis by shorter body length (2.2-2.9 mm vs 4.0 mm in males and 2.9-4.1 mm vs 7.6 mm in females), by the location of the organs of the male reproductive system (from level of the posterior third of the proboscis receptacle in M. brevelemniscus vs in the posterior half of the trunk in M. dakusuiensis), and by length of lemnisci (lemnisci shorter than the proboscis receptacle vs lemnisci longer than the proboscis receptacle). Phylogenetic analyses of almost complete 18S rRNA gene revealed paraphyly of the family Rhadinorhynchidae suggested in previous studies. Micracanthorhynchina dakusuiensis and M. brevelemniscus formed a strongly supported cluster, which formed the earliest diverging branch to the rest of the rhadinorhynchids and transvenids.

Proteocephalid tapeworms of frogs of the family Ranidae ('true' frogs) are reviewed with emphasis on their species diversity, host specificity and geographical distribution. New molecular data (nuclear lsrDNA and mitochondrial COI sequences) are presented for tapeworms of four species of ranid frogs in North America, including the poorly known Ophiotaenia saphena Osler, 1931 of Rana clamitans Latreille and R. catesbeiana (Shaw), which is redescribed using new material from Arkansas, USA. Tapeworms of R. sphenocephala (Cope) and R. pipiens Schreber, the latter previously identified as O. saphena, represent another, putative new species, but are not formally described due to insufficient available material. Proteocephalus papuensis Bursey, Goldberg et Kraus, 2008 from Sylvirana supragrisea (Menzies) is transferred to Ophiotaenia La Rue, 1911 as a new combination. After a critical review of the literature, only nine nominal species of Ophiotaenia are recognised as valid, which is in contrast to the large number of ranid frogs (> 440 spp.). The reasons for this striking disparity are briefly discussed, and a key based on morphology is presented for the identification of all species of Ophiotaenia from the Ranidae. Molecular data are available for only two taxa from North America that form a monophyletic group. The relationships among tapeworms of ranid frogs occurring in other zoogeographical regions are not yet known. The taxonomic status of Batrachotaenia Rudin, 1917, which was erected to accommodate proteocephalids from amphibians, is also discussed. To facilitate future studies, a tabulated summary of all 32 species of proteocephalids belonging to three genera reported from amphibians (frogs and salamanders) is presented, with information on their hosts, distribution, and taxonomically important characters, including key measurements.

Toxoplasma gondii Nicolle et Manceaux, 1909, the etiologic agent of toxoplasmosis, was considered a clonal population with three distinct genetic lineages (I, II and III); however, sequence analysis of different strains has revealed distinct atypical genotypes. Macrophages are essential for immunity against toxoplasmosis and differential cell regulation may affect the course of the disease. In this context, our study aims to investigate the infection by TgChBrUD2, a highly virulent atypical Brazilian strain of T. gondii, on the activation and polarisation of human macrophages. Human macrophage-like cells obtained from THP-1 cells were infected with TgChBrUD2, RH or ME49 strains of T. gondii to evaluate the impact of parasite infection on macrophage polarisation. Our results indicate that the TgChBrUD2 and ME49 strains of T. gondii induced a classic activation of human macrophages, which was confirmed by the high rate of spindle-shaped macrophages, low amount of urea and increase in the levels of nitrite, as well as the down-regulation of M2-markers. In contrast, RH strain promoted an alternative activation of macrophages. The polarisation of human macrophages towards an M1 subtype mediated by TgChBrUD2 and ME49 strains resulted in a low parasite burden, with high levels of IL-6 and MIF. Finally, the M2 subtype triggered by the RH strain culminated in a lower intracellular proliferation index. We concluded that the atypical (TgChBrUD2) and clonal (ME49) strains are able to elicit an M1 subtype, which results in parasitism control, partially explained by the high levels of IL-6 and MIF produced during the infection by these genotypes. In contrast, the clonal (RH) strain promoted a macrophage polarisation towards an M2 subtype, marked by a high parasite burden, with a weak modulation of pro-inflammatory cytokines. Thus, atypical strains can present different mechanisms of pathogenicity and transmissibility compared to clonal strains, as well as they can use distinct strategies to evade the host's immune response and ensure their survival.

In the present study, we have investigated the role of antimalarial drug halofantrine (HF) in inducing the sterile protection against challenges with sporozoites of the live infectious Plasmodium yoelii (Killick-Kendrick, 1967) in Swiss mice malaria model. We observed that during the first to third sequential sporozoite inoculation cycles, blood-stage patency remains the same in the control and chemoprophylaxis under HF drug cover (CPS-HF) groups. However, a delayed blood-stage infection was observed during the fourth and fifth sporozoite challenges and complete sterile protection was produced following the sixth sporozoite challenge in CPS-HF mice. We also noticed a steady decline in liver stage parasite load after 3th to 6th sporozoite challenge cycle in CPS-HF mice. CPS-HF immunisation results in a significant up-regulation of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-12 and iNOS) and down-regulation of anti-inflammatory cytokines (IL-10 and TGF-β) mRNA expression in hepatic mononuclear cells (HMNC) and spleen cells in the immunised CPS-HF mice (after 6th sporozoite challenge) compared to control. Overall, our study suggests that the repetitive sporozoite inoculation under HF drug treatment develops a strong immune response that confers protection against subsequent challenges with sporozoites of P. yoelii.

Based on light and electron microscopical studies, a new nematode parasite, Echinocephalus inserratus sp. n. (Spirurida: Gnathostomatidae), is described from the spiral valve of the broad cowtail stingray Pastinachus ater (Macleay) (Dasyatidae, Myliobatiformes) from off New Caledonia. The new species is morphologically and biometrically most similar to Echinocephalus overstreeti Deardorff et Ko, 1983, differing from it mainly in the absence of serrations on the posterior parts of pseudolabia and on interlabia, and in having a longer gubernaculum (150-299 µm long). Morphologically unidentifiable, mostly encapsulated larvae of Echinocephalus spp. were recorded from the following six species of teleost fishes collected in New Caledonian waters, serving as paratenic hosts: Perciformes: Acanthopagrus berda (Forsskål) (Sparidae) and Nemipterus furcosus (Valenciennes) (Nemipteridae); Tetraodontiformes: Abalistes stellatus (Anonymous), Pseudobalistes fuscus (Bloch et Schneider) (both Balistidae), Lagocephalus sceleratus (Gmelin) (Tetraodontidae) and Aluterus monoceros (Linnaeus) (Monacanthidae). Co-parasitising larvae of Ascarophis sp. and Hysterothylacium sp. were also collected from P. fuscus. All these findings represent new host and geographical records. A key to valid species of Echinocephalus Molin, 1858 is provided.

Taxonomic issues within Trypanorhyncha, e.g., the inaccurate light microscopic visualisation of the hook patterns, are solvable by confocal laser scanning microscopy (CLSM). We applied CLSM imaging to study Trygonicola macropora (Shipley et Hornell, 1906) and Dollfusiella michiae (Southwell, 1929) from Neotrygon caeruliopunctata Last, White et Séret from Bali, Indonesia. To illustrate the strength and limitations of CLSM, images of Otobothrium cysticum (Mayer, 1842) and Symbothriorhynchus tigaminacantha Palm, 2004, both permanent mounts from a collection, were also processed. The CLSM created image stacks of many layers, and edited with IMARIS Software, these layers resulted in three-dimensional images of the armature patterns and internal organs of both species. BABB (benzylalcohol and benzylbenzuolate) clearing was applied to T. macropora. We conclude that trypanorhynch cestodes stained with Mayer-Schuberg's acetic carmine permanently mounted in Canada balsam are suitable for CLSM, allowing detailed analyses of museum type-material as well as freshly collected and processed worms. BABB resulted in imaging the testes in detail, suggesting other stains to be used for CLSM in trypanorhynch cestode research. Application of CLSM for studies of other cestode groups is highly recommended.

Over the last two decades my colleagues and I have assembled the literature on a good percentage of most of the coccidians (Conoidasida) known, to date, to parasitise: Amphibia, four major lineages of Reptilia (Amphisbaenia, Chelonia, Crocodylia, Serpentes), and seven major orders in the Mammalia (Carnivora, Chiroptera, Lagomorpha, Insectivora, Marsupialia, Primates, Scandentia). These vertebrates, combined, comprise about 15,225 species; only about 899 (5.8%) of them have been surveyed for coccidia and 1,946 apicomplexan valid species names or other forms are recorded in the literature. Based on these compilations and other factors, I extrapolated that there yet may be an additional 31,381 new apicomplexans still to be discovered in just these 12 vertebrate groups. Extending the concept to all of the other extant vertebrates on Earth; i.e. lizards (6,300 spp.), rodents plus 12 minor orders of mammals (3,180 spp.), birds (10,000 spp.), and fishes (33,000 spp.) and, conservatively assuming only two unique apicomplexan species per each vertebrate host species, I extrapolate and extend my prediction that we may eventually find 135,000 new apicomplexans that still need discovery and to be described in and from those vertebrates that have not yet been examined for them! Even doubling that number is a significant underestimation in my opinion.

An outbreak of trichinellosis occurred in Stari Banovci, a settlement in the municipality of Stara Pazova, Srem, Republic of Serbia, in March-April 2019. A total of 28 persons were exposed and trichinellosis was confirmed in 24 of them. This outbreak involved members of eight families, their relatives and friends. The infection, due to Trichinella spiralis (Owen, 1835), was associated with consumption of pork sausages procured in a local butcher's shop. The trace-back study revealed that the meat originated from swine that was raised on a small farm belonging to the owner of the shop, who did not have permission from the Veterinary Directorate for slaughtering animals and who put on the market sausages prepared from uninspected meat. Trichinellosis was accompanied by typical clinical symptoms. However, the unusual occurrence of high percentage of pulmonary complications was noticed. The description of this outbreak indicates that medical practitioners should initiate treatment immediately in cases of high suspicion of trichinellosis, even if the serology is negative, to prevent the complications of the disease. In spite of significant achievements in the control of Trichinella infection among domestic pigs and humans in the last 10 years, it is obvious that such cases of breeding animals under inappropriate conditions, slaughtering them without approval and placing uninspected pork on the market represent a source of sporadic outbreaks in Serbia.