A new species of the genus Plagiorhynchus Lühe, 1911 from the intestine of the long-billed curlew (Numenius americanus) from northern Mexico is described. Plagiorhynchus (Plagiorhynchus) aznari n. sp. is morphologically distinguished from other congeneric species from the Americas by having a trunk expanded anteriorly and a cylindrical proboscis, armed with 19 longitudinal rows of hooks, with 14–15 hooks each row. Nearly complete sequences of the small subunit and large subunit of the nuclear ribosomal DNA of the new species were determined and compared with available sequences from GenBank. Phylogenetic analyses inferred from the two molecular markers consistently showed that P. (Plagiorhynchus) aznari n. sp. is closely related to P. (Plagiorhynchus) allisonae, and this clade is sister to a clade formed by P. (Prosthorhynchus) transversus and P. (Prosthorhynchus) cylindraceus from Plagiorhynchidae. The new species represents the second record of the genus in Mexico and the fourth species in the Americas. The phylogenetic relationships among the members of the order Polymorphida in this study provide significant insights into the evolution of ecological associations between parasites and their definitive hosts. Our analyses suggest that the colonization of marine mammals, fish-eating birds and waterfowl in Polymorphidae might have occurred independently, from a common ancestor of Centrorhynchidae and Plagiorhynchidae that colonized terrestrial birds and mammals.

Nematodes of the genus Gongylonema infect a wide range of mammals worldwide but are only sporadically reported in humans. We describe a case of human infection with Gongylonema pulchrum in a 41-year-old man. The patient extracted the nematode from the submucosa under his tongue and correctly self-diagnosed the infection with the help of the Google search engine. In the laboratory, the collected nematode was confirmed as G. pulchrum microscopically by morphological analysis and genetically by amplifying and sequencing the parasite's rDNA. This is the first report of human G. pulchrum infection in Slovenia.

A systematic review was conducted focusing on the distribution of Fasciola species and their snail intermediate hosts (IHs) in East and Southern Africa. The reviewed literature showed that both Fasciola hepatica and F. gigantica are present in East and Southern Africa, and infect a wide range of domestic and wild ruminants. Fasciola gigantica was reported in six East African and five Southern African countries, where Radix natalensis (found in low altitudes) was reported to be the main IH. Fasciola hepatica was reported in Tanzania and Ethiopia (East Africa), and in South Africa and Zimbabwe (Southern Africa), where Galba truncatula (found in high altitudes) was documented as the IH in all countries except in Zimbabwe. Both Fasciola species were documented in Tanzania, Ethiopia, Zimbabwe and South Africa. An overlap of the two was observed in areas with an intermediate altitude in Ethiopia and South Africa, where Pseudosuccinea columella was widespread and assumed to transmit both species. Pseudosuccinea columella has been reported in South Africa and Namibia, and proven to transmit F. gigantica in South Africa; its role in Namibia in the transmission of Fasciola species has not been reported. Other lymnaeid species such as R. rubiginosa were reported in South Africa, and R. auricularia in South Africa and Botswana; their role in the transmission of Fasciola species has not been proven. Future studies should aim to determine the role of P. columella in the geographical spread of the two species in East and Southern African countries.

Despite several studies conducted to determine the genotypes of cystic echinococcosis (CE) agents in humans and other intermediate hosts, the relationship between parasite genotype and clinical presentation of hydatidosis is yet to be well defined. The aim of this study was to compare the genotypes/haplotypes of CE agents of human extra-hepatopulmonary hydatid cysts and common hydatid cysts of the liver. A comparative analysis was carried out between partial cox1 sequences of ten extra-hepatopulmonary hydatid cysts, two liver cysts and reliable sequences from the GenBank database. All the studied hydatid cysts had the Echinococcus granulosus sensu stricto (G1-G2-G3) genotypes. The liver CE cysts were caused by common G1 genotype, while six of the extra-hepatopulmonary cysts had genotypes different from common G1 cysts. The sequences of these six isolates were identical to the G2 and G3 genotypes of E. granulosus sensu stricto; the kidney and peritoneum cysts and most of the brain cysts were identified as G2 genotype, while G3 genotype was only reported in a cyst belonging to the pelvic region. Given the observed differences between the sequences of hydatid cysts, it seems that the replacement of hydatid cysts in organs other than the liver and lungs can be related to their genotypes and probably intra-genotypic characteristics. It was hypothesized that in each geographic area, less frequent genotypes were likely to be more consistent with placement in the host's unusual organs.

Helminths were examined from 145 scaled quail (Callipepla squamata) collected during the 2012–2013, 2013–2014 and 2014–2015 hunting seasons from a semi-arid region of Texas that spans four ecoregions. Helminth infracommunities were species poor, averaging 1.7 (range 1–4) species. Six species occurred within the component community of which one (Oxyspirura petrowi) is known to be pathogenic to quail. Aulonocephalus pennula was most abundant (9991 individuals, 95% of total) followed by O. petrowi (391 individuals, 4%). Each of the remaining four species was rare (≤21% prevalence) and contributed few individuals (<1%). In the High Plains ecoregion, prevalence of O. petrowi was higher in host collections made during the 2013–2014 hunting season than either hunting seasons 2012–2013 or 2014–2015 and was higher in the High Plains ecoregion than the Edwards Plateau ecoregion during the 2013–2014 hunting season. Mean abundance of A. pennula and O. petrowi was higher in scaled quail from the High Plains ecoregion than the Edwards Plateau ecoregion. Our results provide new information about helminth fauna in scaled quail, persistence of indirect lifecycle helminth species within a semi-arid region, and the occurrence of pathogenic helminth species within this host species.

The trematodes from South American reptiles are poorly known, with only one life cycle completely characterized. We used molecular and morphological methods to characterize diplostomoid metacercariae found in 29 of 86 pointedbelly frogs, Leptodactylus podicipinus (Cope, 1862) collected in a marsh pond in Selvíria, in the central-west region of Brazil. The metacercariae were identified as Heterodiplostomum lanceolatum Dubois, 1936 (Proterodiplostomidae), a rarely reported species that matures in snakes. In phylogenetic analysis of partial sequences from 28S rDNA, H. lanceolatum fell within a polytomy with the proterodiplostomid Crocodilicola pseudostoma (molecular divergence of 4.1%) and other members of the superfamily Diplostomoidea. Our collections provide insights into the ecology of this parasite, in that infected frogs were smaller than uninfected frogs, and metacercariae were more numerous in the abdominal cavity and hindlimb muscles than in abdominal muscles, which suggests directions for future research on the transmission and pathology of this proterodiplostomid.

Human subcutaneous dirofilariosis has several clinical presentations. Many cases present as subcutaneous nodules, as a consequence of a local inflammatory reaction that encapsulates and destroys the worms. In addition, there are cases in which migrating worms located in the ocular area remain unencapsulated. In the present work, the levels of two pro-inflammatory eicosanoids, thromboxane B2 (TxB2) and leukotriene B4 (LTB4) are analysed by commercial Enzime-Linked immunosorbent assay (ELISA) in serum samples from 43 individuals, 28 diagnosed as having subcutaneous dirofilariasis presenting a subcutaneous nodule, five diagnosed as having dirofilariasis, in which the worms remained unencapsulated in the periphery of the eye, and ten healthy individuals living in a non-endemic area, used as controls. The worms were surgically removed, identifying Dirofilaria repens as the causative agent in all cases, by Polymerase Chain Reaction (PCR). Individuals with nodules showed significantly higher levels of TxB2 and LTB4 than healthy controls, whereas significant differences in LTB4 levels were observed between individuals with unencapsulated worms and healthy controls. It is speculated that the absence of LTB4 may contribute to the fact that worms remain unencapsulated as a part of immune evasion mechanisms.

Nematodes of the genus Physaloptera are globally distributed and more than 100 species are known. Their life cycle involves insects, including beetles, cockroaches and crickets, as intermediate hosts. This study describes a new species of Physaloptera and reports molecular phylogenetic analyses to determine its relationships within the family Physalopteridae. Physaloptera amazonica n. sp. is described from the stomach of the caviomorph rodent Proechimys gardneri collected in the Amazon rainforest in the state of Acre, Brazil. The species is characterized by the male having the first and second pair of sessile papillae asymmetrically placed, lacking a median papilla-like protuberance between the third pairs of sessile papillae, differentiated by size and shape of the spicules, while females have four uterine branches. For both nuclear 18S rRNA and MT-CO1 gene-based phylogenies, we recovered Turgida sequences forming a clade nested within Physaloptera, thus making Physaloptera paraphyletic to the exclusion of Turgida, suggesting that the latter may have evolved from the former monodelphic ancestral state to a derived polydelphic state, or that some species of Physaloptera may belong to different genera. Relationships between most taxa within Physaloptera were poorly resolved in our phylogenies, producing multifurcations or a star phylogeny. The star-like pattern may be attributed to evolutionary processes where past simultaneous species diversification events took place. Physaloptera amazonica n. sp. formed an independent lineage, separately from the other species of Physaloptera, thus supporting the status of a new species. However, all molecular data suggested a closer relationship with other Neotropical species. In conclusion, we added a new species to this already largely diverse genus Physaloptera, bringing new insights to its phylogenetic relationships. Further analyses, adding more species and markers, should provide a better understanding of the evolutionary history of physalopterids.

The diversity and importance of Echinococcus species in domesticated animals in Kazakhstan are poorly understood. In this study, 17 cysts of Echinococcus were collected from cattle and a further 17 cysts from sheep. DNA was extracted from the individual cysts and used for polymerase chain reaction amplification of mitochondrial subunit 1 of the cox1 and nadh1 gene. Amplicon sequencing results revealed the presence of Echinococcus granulosus sensu stricto G1 in 15 cattle and 15 sheep, and G3 genotype from two cattle. Echinococcus canadensis (G6/G7 strain) was found in two cysts originating from sheep.

Toxocara canis is an important zoonotic roundworm distributed worldwide. The infective larvae of T. canis are one of the causes of visceral larva migrans (VLM), a clinical syndrome in humans. Diagnosing VLM is difficult, and the differential diagnosis of the larval development stage is limited. Therefore, this experimental research aimed to diagnose T. canis larvae using a molecular method, not only in liver tissue, which is the most commonly affected tissue, but also in the limb muscles, lungs and brain tissues. For this purpose, 24 BALB/c mice were infected with 1000 embryonated T. canis eggs. Necropsies were performed on the second, fourth, seventh and 14th days post-infection. While a part of the samples were digested with pepsin-HCl, the molecular method was used for the remainder of the samples to replicate the mitochondrial DNA adenosine triphosphate (ATP) synthase subunit-6 gene region of T. canis. BbsI, a restriction endonuclease, was used to determine the specificity of the amplicons obtained from Polymerase chain reaction (PCR). The detection limit for embryonated eggs was recorded. The PCR results showed that the sensitivity of the PCR analysis was 83.3% in the liver (with 88.8% accuracy), 87.5% in the lungs (with 91.6% accuracy) and 75.0% in the brain, forelimb and hindlimb muscles (with 83.3% accuracy). In all tissues, the test specificity was determined to be 100%. In this study, the molecular method was applied to only experimentally infected BALB/c mice tissues; thus, it is suggested that it can be also employed in different paratenic hosts and materials possibly infected with T. canis.

Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.

Schistosomiasis is an inflammatory disease that occurs when schistosome species eggs are deposited in the liver, resulting in fibrosis and portal hypertension. Schistosomes can interact with host inflammasomes to elicit host immune responses, leading to mitochondrial damage, generation of high levels of reactive oxygen species (ROS) and activation of apoptosis during inflammation. This study aims to examine whether ROS and NF-κB (p65) expression elicited other types of inflammasome activation in Schistosoma mansoni-infected mouse livers. We examine the relationship between inflammasome activation, mitochondrial damage and ROS production in mouse livers infected with S. mansoni. We demonstrate a significant release of ROS and superoxides and increased NF-κB (p65) in S. mansoni-infected mouse livers. Moreover, activation of the NLRP3 and AIM2 inflammasomes was triggered by S. mansoni infection. Stimulation of HuH-7 hepatocellular carcinoma cells with soluble egg antigen induced activation of the AIM2 inflammasome pathway. In this study, we demonstrate that S. mansoni infection promotes both NLRP3 and AIM2 inflammasome activation.

Digeneans (Trematoda: Digenea) are endoparasites that present a complex life cycle, generally involving an intermediate invertebrate host and a vertebrate host. There is limited information about which species of molluscs may act as intermediate hosts in the upper Paraná River floodplain (UPRF), where Aylacostoma chloroticum can be considered a potential candidate. The study of digeneans in this region is important because some of these parasites are potentially zoonotic, and, therefore, are relevant to public health. However, the correct identification of these organisms during the larval stages is difficult because of the lack of morphologically diagnostic characteristics. The objective of this study was to identify and molecularly characterize the larval stages of digeneans found in A. chloroticum in the UPRF, using the mitochondrial marker of subunit I of cytochrome c oxidase and the 28S nuclear marker. The molluscs were examined in the laboratory and three morphotypes of cercariae were found. DNA was extracted from the specimens obtained and was then amplified and sequenced. The morphotypes exhibited high genetic similarities with Pseudosellacotyla, Paralecithodendrium and Philophthalmus, indicating that these organisms belong to these genera. This is the first record of larval stages of these genera in molluscs collected in the UPRF.

Assessing fish stocks has important implications for fisheries management and conservation biology. Gurnards are marine demersal fish that commonly occur in the Mediterranean, but their population in this region remains to be quantified. This study examines the population structure of the longfin gurnard Chelidonichthys obscurus (Walbaum, 1792) in waters off eastern Tunisia, using morphometry and parasite assemblages. A total of 134 fish are investigated from two studied zones – the Gulf of Hammamet and the Gulf of Gabès. Discriminant analysis is used to compare gurnard populations in the two studied zones using 13 morphometric characters and the infection parameters of seven parasites. Morphometric analysis reveals strong spatial variations between the studied zones, providing evidence for the existence of an ecological differentiation along the eastern Tunisian coast. Mahalanobis distances show that body height, pectoral fin length and first dorsal fin length are the most salient morphometric characters for determining the position of samples from the Gulf of Gabès. The effectiveness of using parasites to study longfin gurnard stocks is uncertain. The use of short-lived parasites as biological tags is questionable, at least in the present case. Future research, based on complementary approaches such as otolith microchemistry and genetics, may improve our understanding of the global stock structure of longfin gurnard to suitably inform regional organizations involved in fisheries management.

Interspecific interactions between parasites sharing the same host are often antagonistic; the presence of one species decreases the number of individuals or negatively affects both the distribution and reproduction of the other species. Antagonistic interactions between co-infecting parasites may translate into direct competition or interactive segregation, but elements of both may be present. Potential interactions between two acanthocephalan species, Pomphorhynchus laevis and Acanthocephalus anguillae, were studied in the field in two of their natural fish definitive hosts. There was no evidence for competitive exclusion between P. laevis and A. anguillae. However, a negative interaction was found for the first time in the field between these two species. Based on the analysis of parasite abundance and total biomass using a static regression approach, I found that the abundance and total biomass of parasite was also limited by host characteristics. These results are consistent with previous laboratory studies on competition between P. laevis and A. anguillae.

Dracunculiasis is the first parasitic disease set for eradication. However, recent events related to the Dracunculus medinensis epidemiology in certain African countries are apparently posing new challenges to its eradication. Two novel facts have emerged: the existence of animal reservoirs (mainly dogs but also cats and baboons), and possibly a new food-borne route of transmission by the ingestion of paratenic (frogs) or transport (fish) hosts. Therefore, instead of being exclusively a water-borne anthroponosis, dracunculiasis would also be a food-borne zoonosis. The existence of a large number of infected dogs, mainly in Chad, and the low number of infected humans, have given rise to this potential food-borne transmission. This novel route would concern not only reservoirs, but also humans. However, only animals seem to be affected. Dracunculus medinensis is on the verge of eradication due to the control measures which, classically, have been exclusively aimed at the water-borne route. Therefore, food-borne transmission is probably of secondary importance, at least in humans. In Chad, reservoirs would become infected through the water-borne route, mainly in the dry season when rivers recede, and smaller accessible ponds, with a lower water level containing the infected copepods, appear, whilst humans drink filtered water and, thus, avoid infection. The total absence of control measures aimed at dogs (or at other potential reservoirs) up until the last years, added to the stimulating reward in cash given to those who find parasitized dogs, have presumably given rise to the current dracunculiasis scenario in Chad.

This study assessed the in vitro anthelmintic (AH) activity of methanol and acetone:water leaf extracts from Annona squamosa, A. muricata and A. reticulata against Haemonchus contortus eggs. The egg hatch test was used to determine the effective concentrations required to inhibit 50% of eggs hatching (EC50). The role of polyphenols on AH activity was measured through bioassays with and without polyvinylpolypyrrolidone (PVPP). Methanolic extracts mainly caused the death of eggs at the morula stage (ovicidal activity). Meanwhile, acetone:water extracts caused egg-hatching failure of developed larvae (larvae failing eclosion (LFE) activity). The lowest EC50 values against H. contortus eggs were observed for the methanolic extracts from A. reticulata and A. muricata (274.2 and 382.9 µg/ml, respectively). From the six extracts evaluated, the methanolic extracts of A. muricata, A. reticulata and A. squamosa showed the highest ovicidal activity, resulting in 98.9%, 92.8% and 95.1% egg mortality, respectively. When the methanolic extract of A. squamosa was incubated with PVPP, its AH activity increased. Similarly, when acetone:water extracts of A. muriata and A. reticulata were incubated with PVPP, their LFE activity increased. Alkaloids were only evident in methanolic extracts, irrespective of PVPP incubation. The presence of acetogenins was not observed. In conclusion, methanolic extracts obtained from leaves of A. muricata, A. reticulata and A. squamosa showed ovicidal activity affecting the morula of H. contortus eggs, with minor LFE activity. Meanwhile, acetone:water extracts showed mostly LFE activity, with a lower proportion of ovicidal activity.

A central and critical step in the molecular detection of soil-transmitted helminths from environmental sources is the extraction of DNA from the eggs. In this study, we investigated the yield of DNA extracted from known quantities (500, 100, 50, 20, 10 and 5) of Ascaris suum eggs, as well as directly from wastewater and sludge samples containing Ascaris spp. eggs, using six commercial DNA extraction kits. The amount of DNA extracted was quantified with NanoDrop, Qubit and Ct values from quantitative polymerase chain reaction (qPCR) assay using CFX96 Touch™ real-time PCR equipment. The PowerLyzer Ultraclean Microbial DNA isolation kit and PowerSoil DNA isolation kit gave the highest yield of DNA based on the NanoDrop, Qubit and Ct values. However, the qPCR results indicate that in some of the kits, PCR inhibitors may have been carried over to the PCR reaction. DNA extraction kits that incorporate a bead-beating step as well as other mechanical eggshell disruption steps were superior in extracting DNA from Ascaris spp. eggs. Additionally, for the accurate quantification of extracted DNA, the use of Ct values from qPCR and Qubit readings gives better results compared to the NanoDrop readings. For efficient downstream applications, the use of DNA extraction kits with superior inhibitor removal technology is essential, in addition to a high yield of DNA.

Intraspecific diversity in parasites with heteroxenous life cycles is guided by reproduction mode, host vagility and dispersal, transmission features and many other factors. Studies of these factors in Digenea have highlighted several important patterns. However, little is known about intraspecific variation for digeneans in the marine Arctic ecosystems. Here we analyse an extended dataset of partial cox1 and nadh1 sequences for Tristriata anatis (Notocotylidae) and confirm the preliminary findings on its distribution across Eurasia. Haplotypes are not shared between Europe and the North Pacific, suggesting a lack of current connection between these populations. Periwinkle distribution and anatid migration routes are consistent with such a structure of haplotype network. The North Pacific population appears ancestral, with later expansion of T. anatis to the North Atlantic. Here the parasite circulates widely, but the direction of haplotype transfer from the north-east to the south-west is more likely than the opposite. In the eastern Barents Sea, the local transmission hotspot is favoured.

The co-occurrence of hepatic cystic echinococcosis (CE) and alveolar echinococcosis (AE) is extremely rare. Here, we present the clinical manifestations and treatment outcomes of three cases with co-occurring CE and AE in the liver. Computed tomography (CT), magnetic resonance imaging and 18FFluorodeoxyglucose Positron Emission Tomography-CT were used for preoperative diagnosis. Specimens were taken intraoperatively and sent for pathological studies to confirm the coexistence of CE and AE by laminated membrane, daughter cysts or germinal layer and infiltration structure. Albendazole was prescribed after operation for 12 months. All patients were completely recovered and showed no recurrence at last follow-up. Therefore, surgical intervention and postoperative application of albendazole are recommended for patients with concurrence of hepatic AE and CE.