Showing papers in "Journal of Helminthology in 2019"

Testing the higher-level phylogenetic classification of Digenea (Platyhelminthes, Trematoda) based on nuclear rDNA sequences before entering the age of the 'next-generation' Tree of Life

Abstract: Digenea Carus, 1863 represent a highly diverse group of parasitic platyhelminths that infect all major vertebrate groups as definitive hosts. Morphology is the cornerstone of digenean systematics, but molecular markers have been instrumental in searching for a stable classification system of the subclass and in establishing more accurate species limits. The first comprehensive molecular phylogenetic tree of Digenea published in 2003 used two nuclear rRNA genes (ssrDNA = 18S rDNA and lsrDNA = 28S rDNA) and was based on 163 taxa representing 77 nominal families, resulting in a widely accepted phylogenetic classification. The genetic library for the 28S rRNA gene has increased steadily over the last 15 years because this marker possesses a strong phylogenetic signal to resolve sister-group relationships among species and to infer phylogenetic relationships at higher levels of the taxonomic hierarchy. Here, we have updated the database of 18S and 28S rRNA genes until December 2017, we have added newly generated 28S rDNA sequences and we have reassessed phylogenetic relationships to test the current higher-level classification of digeneans (at the subordinal and subfamilial levels). The new dataset consisted of 1077 digenean taxa allocated to 106 nominal families for 28S and 419 taxa in 98 families for 18S. Overall, the results were consistent with previous higher-level classification schemes, and most superfamilies and suborders were recovered as monophyletic assemblages. With the advancement of next-generation sequencing (NGS) technologies, new phylogenetic hypotheses from complete mitochondrial genomes have been proposed, although their power to resolve deep levels of trees remains controversial. Since data from NGS methods are replacing other widely used markers for phylogenetic analyses, it is timely to reassess the phylogenetic relationships of digeneans with conventional nuclear rRNA genes, and to use the new analysis to test the performance of genomic information gathered from NGS, e.g. mitogenomes, to infer higher-level relationships of this group of parasitic platyhelminths.

Echinococcus granulosus genotypes in Iran: a systematic review.

Abstract: Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu lato (s.l.) is a significant zoonosis, especially in developing countries of the Middle East, with many studies focusing on CE genotypes in Iran. We performed a systematic review to determine the exact status of E. granulosus genotypes in the country. We explored English (Pubmed, Scopus, ISI Web of Science and Science Direct) and Persian (Magiran, Iran Medex and Scientific Information Database) databases along with Google Scholar. Our review included 73 studies published prior to the end of 2015. In total, 2952 animal (intermediate and definitive) hosts were examined, and the prevalent genotypes comprised G1 (92.75%) and G6 (4.53%) in sheep, cattle, camels, goats and buffaloes; G3 (2.43%) in five herbivore hosts and dogs; G7 (0.2%) in sheep and goats; and G2 (0.06%) in dogs. G1 was mostly dominant in West Azerbaijan, whereas G3 and G6 were identified most frequently in the provinces of Isfahan and Fars, respectively. Regarding human CE infection, 340 cases were reported from Iran, with the identified genotypes G1 (n = 320), G6 (n = 13) and G3 (n = 7). Most CE-infected humans originated from Isfahan province (168 cases), whereas the lowest number of infected persons was noted in Kerman province (two cases). The information obtained from this systematic review is central to better understanding the biological and epidemiological characteristics of E. granulosus s.l. genotypes in Iran, leading to more comprehensive control strategies.

The first report of Xenorhabdus indica from Steinernema pakistanense: co-phylogenetic study suggests co-speciation between X. indica and its steinernematid nematodes

Abstract: During a survey in agricultural fields of the sub-humid region of Meerut district, India, two strains of entomopathogenic nematodes, labelled CS31 and CS32, were isolated using the Galleria baiting technique. Based on morphological and morphometric studies, and molecular data, the nematodes were identified as Steinernema pakistanense, making this finding the first report of this species from India. For the first time, we performed a molecular and biochemical characterization of the bacterial symbiont of S. pakistanense. Furthermore, a co-phylogenetic analysis of the bacteria from the monophyletic clade containing a symbiont of S. pakistanense, together with their nematode hosts, was conducted, to test the degree of nematode–bacteria co-speciation. Both isolates were also tested in a laboratory assay for pathogenicity against two major pests, Helicoverpa armigera and Spodoptera litura. The morphology of the Indian isolates corresponds mainly to the original description, with the only difference being the absence of a mucron in first-generation females and missing epiptygmata in the second generation. The sequences of bacterial recA and gyrB genes have shown that the symbiont of S. pakistanense is closely related to Xenorhabdus indica, which is associated with some other nematodes from the ‘bicornutum’ group. Co-phylogenetic analysis has shown a remarkable congruence between the nematode and bacterial phylogenies, suggesting that, in some lineages within the Steinernema / Xenorhabdus complex, the nematodes and bacteria have undergone co-speciation. In the virulence assay, both strains caused a 100% mortality of both tested insects after 48 h, even at the lowest doses of 25 infective juveniles per insect, suggesting that S. pakistanense could be considered for use in the biocontrol of these organisms in India.

Stable-isotope analysis: a neglected tool for placing parasites in food webs.

Abstract: Parasites are often overlooked in the construction of food webs, despite their ubiquitous presence in almost every type of ecosystem. Researchers who do recognize their importance often struggle to include parasites using classical food-web theory, mainly due to the parasites' multiple hosts and life stages. A novel approach using compound-specific stable-isotope analysis promises to provide considerable insight into the energetic exchanges of parasite and host, which may solve some of the issues inherent in incorporating parasites using a classical approach. Understanding the role of parasites within food webs, and tracing the associated biomass transfers, are crucial to constructing new models that will expand our knowledge of food webs. This mini-review focuses on stable-isotope studies published in the past decade, and introduces compound-specific stable-isotope analysis as a powerful, but underutilized, newly developed tool that may answer many unresolved questions regarding the role of parasites in food webs.

Meloidogyne daklakensis n. sp. (Nematoda: Meloidogynidae), a new root-knot nematode associated with Robusta coffee (Coffea canephora Pierre ex A. Froehner) in the Western Highlands, Vietnam.

Abstract: The root-knot nematode species Meloidogyne daklakensis n. sp. was discovered on the roots of Robusta coffee (Coffea canephora Pierre ex A. Froehner) in Dak Lak Province, Vietnam. This species is characterized by the females having rounded or oval perineal patterns, smooth, regular, continuous striae, and reduced lateral lines. The dorsal arch is low, rounded and encloses a quite distinct vulva and tail tip. The stylet is normally straight with well-developed and posteriorly sloped knobs. The males have a rounded cap that extends posteriorly into the lip region. The procorpus is outlined distinctly, and is three times longer than the metacorpus. The metacorpus is ovoid, with a strong valve apparatus. The species closely resembles M. marylandi, M. naasi, M. ovalis, M. panyuensis, M. lopezi, M. mali and M. baetica in the perineal pattern of the females, and the morphology of the males and the second-stage juveniles. Nonetheless, it can be differentiated from other species by a combination of morphometric, morphological and molecular characteristics. Phylogenetic analysis was conducted based on the internal transcribed spacer (ITS) and 28S rDNA as well as the region between the cytochrome c oxidase I (COI) and cytochrome c oxidase II (COII) mitochondrial genes. Herein, this nematode is described, illustrated, and designated as a new species, Meloidogyne daklakensis sp. n., based on morphometric, morphological and molecular analyses.

Metacommunity structure of the helminths of the black-eared opossum Didelphis aurita in peri-urban, sylvatic and rural environments in south-eastern Brazil

Abstract: Among the Brazilian marsupials, the species of the genus Didelphis are the most parasitized by helminths. This study aimed to describe the species composition and to analyse the helminth communities of the Atlantic Forest common opossum Didelphis aurita at infracommunity and component community levels using the Elements of Metacommunity Structure Analysis, considering peri-urban, sylvatic and rural environments in the state of Rio de Janeiro, Brazil. Seventy-three specimens of D. aurita were captured during the study. Fourteen species of helminths were collected: nine of the phylum Nematoda (Trichuris minuta, Trichuris didelphis, Globocephalus marsupialis, Heterostrongylus heterostrongylus, Travassostrongylus orloffi, Viannaia hamata, Aspidodera raillieti, Cruzia tentaculata and Turgida turgida); four of the phylum Platyhelminthes (three species of Trematoda, Duboisiella proloba, Brachylaima advena and Rhopalias coronatus, and one species of the class Cestoda); and one species of the phylum Acanthocephala (Oligacanthorhynchus microcephalus). More than 95% of the animals were infected with parasites. The most abundant parasite species were V. hamata, T. turgida, C. tentaculata and T. orloffi. The nematodes T. turgida, C. tentaculata and A. raillieti were the most dominant species in the three environments. The analysis of the metacommunity structure indicated a structured pattern as a function of the environmental gradient, but only when all localities were considered together. At the infracommunity and component community levels, quasi-nested and nested structures with stochastic species loss were observed, respectively. The results indicate that the settlement of helminth species in the opossums can be attributed to the heterogeneity among individual hosts in relation to their exposure to parasites and to extrinsic factors, which vary among habitats, considering a regional scale. On a local scale, the characteristics of this host species may promote homogenization among individual hosts in terms of their exposure to the helminths. This study constitutes a new host record for the species G. marsupialis and T. didelphis and represents a pioneering investigation of the helminth metacommunity of a Neotropical marsupial.

Best practice guidelines for studies of parasite community ecology

Abstract: In recent decades, parasite community ecology has produced hundreds of studies on an ever-growing number of host species, and developed into an active sub-discipline of parasitology. However, this growth has been characterized by a lack of standards in the practices used by researchers, with many common approaches being flawed, unjustified or misleading. Here, in the hope of promoting advances in the study of parasite community ecology, I identify some of the most common errors or weaknesses in past studies, and propose ten simple rules for best practice in the field. They cover issues including, among others, taxonomic resolution, proper and justifiable analytical methods, higher-level replication, controlling for sampling effort or species richness, accounting for spatial distances, using experimental approaches, and placing raw data in the public domain. While knowledge of parasite communities has expanded in breadth, with more and more host species being studied, true progress has been very limited with respect to our understanding of fundamental general processes shaping these communities. It is hoped that the guidelines presented here can direct researchers away from the entrenched use of certain approaches flawed in design, analysis or interpretation, by offering a more rigorous and standardized set of practices, and, hopefully, a way forward.

Helminth endoparasites of the smooth newt Lissotriton vulgaris: linking morphological identification and molecular data.

Abstract: The helminth endoparasites of many European amphibian species are often known exclusively from morphological descriptions. A molecular library of DNA sequence data linked to morphological identifications is still in its infancy. In this paper, we aim to contribute to such a library on the smooth newt Lissotriton vulgaris, the intermediate and definitive host of 31 helminth parasites, according to evidence published so far. Newts (n = 69) were collected at two study sites in western Germany and examined for the presence of helminths. A total of five helminth species were detected in 56 (81%) of the newts, but only one or two species infected a single host. Four out of five helminth species were identified morphologically and based on DNA sequences as Parastrigea robusta (metacercariae), Oswaldocruzia filiformis, Megalobatrachonema terdentatum (adults and larvae) and Cosmocerca longicauda, and the corresponding sequences were provided subsequently. Oswaldocruzia molgeta was confirmed to be a junior synonym of O. filiformis. Molecular data on a fifth species (a cosmocercid nematode) that could not be identified at species level were added to GenBank. These findings increased the molecular library on morphologically identified smooth newt parasites significantly, from 12 to 15 entries.

Schistosomicidal, antifibrotic and antioxidant effects of Cucurbita pepo L. seed oil and praziquantel combined treatment for Schistosoma mansoni infection in a mouse model.

Abstract: Despite the seriousness of schistosomiasis, its treatment depends only on praziquantel (PZQ), which has begun to lose its efficacy against the emergent Schistosoma mansoni-resistant strains. Therefore, the discovery of a novel schistosomicidal drug is an urgent priority. This study was designed to evaluate treatment with Cucurbita pepo L. (pumpkin) seed oil (PSO) alone and combined with PZQ against S. mansoni in experimentally infected mice. The study involved five groups: GI was the normal control; GII was the infected control; GIII was treated with an oral dose of PZQ of 500 mg/kg/day for two successive days, starting in the sixth week post infection; GIV was treated with an oral dose of PSO of 50 mg/kg/day for four weeks, starting in the fourth week post infection; and GV was treated with combined PSO-PZQ. Worm burden, tissue egg load and oogram pattern were estimated, and the ultrastructure alterations were examined. Histopathological examination of granuloma diameters, collagen deposition (Picro Sirius red stain), and angiogenesis (immunohistochemical expression of CD34+) was conducted and serum liver enzymes were measured to assess the liver condition. Moreover, the oxidative stress was evaluated by determining the amounts of malondialdehyde and superoxide dismutase in liver homogenates. The results revealed significant changes in all the assessed parameters with PSO administration. However, PZQ was significantly more effective as an antiparasitic agent, whereas PSO was better in terms of fibrosis and oxidative stress. The most significant results were obtained in group V, which may be attributed to a synergy between PZQ and PSO, with antiparasitic, antioxidant and antifibrotic properties.

Role of ecology and phylogeny in determining tapeworm assemblages in skates (Rajiformes).

Abstract: An understanding of the mechanisms that determine host and parasite relationships is a central aim in parasitology. Association of a parasite species with a host species may be influenced primarily by phylogenetic constraints that cause parasite species to co-speciate with their host species, or predominantly by ecological parameters that influence all other co-evolutionary scenarios. This study aimed to investigate the role of co-speciation as well as other co-evolutionary scenarios in influencing the assemblages of tapeworm parasites (marine cestodes) in skate hosts (Rajiformes) using a modification of the PACo (Procrustean Approach to Cophylogeny) method. The study found that phylogeny and host ecology are both significant predictors of skate-tapeworm relationships, implying that co-speciation as well as other co-evolutionary scenarios are shaping these associations. The study also investigated the key ecological parameters influencing host-switching and found that host diet, distribution depth, average body size and geographical location have a combined effect. Given the importance of parasites in ensuring healthy and stable marine ecosystems, the findings of this study have implications for conservation management worldwide.

Morphological and molecular characterization of an enigmatic clinostomid trematode (Digenea: Clinostomidae) parasitic as metacercariae in the body cavity of freshwater fishes (Cichlidae) across Middle America.

Abstract: The family Clinostomidae Luhe, 1901 contains 29 species allocated to seven genera, of which Clinostomum Leidy, 1856 is the most diverse, with c. 14 valid species. The diversity of Clinostomum has been assessed, combining morphological and molecular data. The genetic library for species in this genus has increased steadily, although there is little or no information for the other genera included in the family. Molecular phylogenetic relationships among the genera of clinostomids have not been assessed, and their classification is still based on morphological traits. The monotypic Ithyoclinostomum was described from a fish-eating bird in Brazil, and its metacercariae have been found in several locations in South America, parasitizing erythrinid freshwater fishes. We collected unusually large metacercariae from the body cavity of cichlids in several locations across Middle America. These metacercariae exhibited some resemblance to Ithyoclinostomum, although several differences prevent their inclusion in Ithyoclinostomum dimorphum, casting doubt on their taxonomic identification. The main objective of this paper was to characterize the metacercariae collected in cichlids using both morphology and molecular data from three molecular markers, and to assess the molecular phylogenetic relationships among the genera of Clinostomidae to establish the position of the newly generated sequences. We took a conservative position and tentatively placed the metacercariae as belonging to Ithyoclinostomum.

Development of Phasmarhabditis hermaphrodita (and members of the Phasmarhabditis genus) as new genetic model nematodes to study the genetic basis of parasitism

Abstract: The genetic mechanisms of how free-living nematodes evolved into parasites are unknown. Current genetic model nematodes (e.g. Caenorhabditis elegans) are not well suited to provide the answer, and mammalian parasites are expensive and logistically difficult to maintain. Here we propose the terrestrial gastropod parasite Phasmarhabditis hermaphrodita as a new alternative to study the evolution of parasitism, and outline the methodology of how to keep P. hermaphrodita in the lab for genetic experiments. We show that P. hermaphrodita (and several other Phasmarhabditis species) are easy to isolate and identify from slugs and snails from around the UK. We outline how to make isogenic lines using 'semi-natural' conditions to reduce in-lab evolution, and how to optimize growth using nematode growth media (NGM) agar and naturally isolated bacteria. We show that P. hermaphrodita is amenable to forward genetics and that unc and sma mutants can be generated using formaldehyde mutagenesis. We also detail the procedures needed to carry out genetic crosses. Furthermore, we show natural variation within our Phasmarhabditis collection, with isolates displaying differences in survival when exposed to high temperatures and pH, which facilitates micro and macro evolutionary studies. In summary, we believe that this genetically amenable parasite that shares many attributes with C. elegans as well as being in Clade 5, which contains many animal, plant and arthropod parasites, could be an excellent model to understand the genetic basis of parasitism in the Nematoda.

First molecular identification of Strongyloides fuelleborni in long-tailed macaques in Thailand and Lao People's Democratic Republic reveals considerable genetic diversity.

Abstract: Strongyloides fuelleborni is a soil-transmitted nematode parasite of non-human primates. The worm is prevalent also in human populations in Africa and South-East Asia. In this study, we amplified and sequenced a portion of the 18S ribosomal RNA gene (rRNA) and of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of Strongyloides adult males recovered from faecal samples from long-tailed macaques (Macaca fascicularis) in Thailand and Lao PDR. The prevalence in Thailand was 31.1% (55/177) and in Lao PDR it was 62.1% (41/66), with an overall prevalence of 39.5% (96/243). All 18S rRNA sequences that we obtained (n = 96) showed 100% identity with published S. fuelleborni sequences. The 96 cox1 sequences that we obtained represented 32 new haplotypes. When included with the 17 previously known haplotypes from S. fuelleborni, the cox1 sequences fell into four clusters, which had clear geographical structure. This is the first molecular confirmation of S. fuelleborni in long-tailed macaques in Thailand and Lao PDR. Clearly, awareness needs to be raised of the zoonotic potential of S. fuelleborni. A monitoring programme should be organized, taking into account the role of reservoir hosts (i.e. monkeys) in the natural background of human strongyloidiasis caused by S. fuelleborni.

Helminth community and host dynamics in northern bobwhites from the Rolling Plains ecoregion, U.S.A.

Abstract: One hundred and sixty-one northern bobwhites (Colinus virginianus; hereafter ‘bobwhite’) were examined from the Rolling Plains ecoregion of Texas and western Oklahoma from 2011 to 2013. Complete necropsies yielded 13 species, of which two are new host (Gongylonema phasianella) and region (Eucoleus contortus) records and three (Dispharynx nasuta, Tetrameres pattersoni and Oxyspirura petrowi) are known to cause morbidity and mortality. Of the species found, Aulonocephalus pennula commonly occurred, Oxyspirura petrowi was intermediate in prevalence, and the remaining species were rare. Species richness was similar compared to studies from the southeastern U.S., but higher than studies from the same region. In addition, 12 of the 13 species were heteroxenous helminths, supporting the theory that heteroxenous helminths in semi-arid regions are more successful than monoxenous helminths. Prevalence and abundance of A. pennula and O. petrowi were higher in adult bobwhites than in juveniles. Abundance of A. pennula and O. petrowi was higher at southern locations compared to northern locations in the study area. Our study is the first to provide a current assessment of the bobwhite helminth community across the Rolling Plains ecoregion of the U.S.

Checklist of marine mammal parasites in New Zealand and Australian waters.

Abstract: Marine mammals are long-lived top predators with vagile lifestyles, which often inhabit remote environments. This is especially relevant in the oceanic waters around New Zealand and Australia where cetaceans and pinnipeds are considered as vulnerable and often endangered due to anthropogenic impacts on their habitat. Parasitism is ubiquitous in wildlife, and prevalence of parasitic infections as well as emerging diseases can be valuable bioindicators of the ecology and health of marine mammals. Collecting information about parasite diversity in marine mammals will provide a crucial baseline for assessing their impact on host and ecosystem ecology. New studies on marine mammals in New Zealand and Australian waters have recently added to our knowledge of parasite prevalence, life cycles and taxonomic relationships in the Australasian region, and justify a first host-parasite checklist encompassing all available data. The present checklist comprises 36 species of marine mammals, and 114 species of parasites (helminths, arthropods and protozoans). Mammal species occurring in New Zealand and Australian waters but not included in the checklist represent gaps in our knowledge. The checklist thus serves both as a guide for what information is lacking, as well as a practical resource for scientists working on the ecology and conservation of marine mammals.

According to mitochondrial DNA evidence, Parascaris equorum and Parascaris univalens may represent the same species.

Abstract: Parascarosis is caused mainly by parasitic infections with Parascaris equorum and Parascaris univalens, the most common ascarid nematodes, in the small intestine of equines. Parascarosis often causes severe illness and even death in foals and yearlings. In this study, we obtained the complete sequence of the P. equorum mitochondrial (mt) genome and compared its organization and structure with that of P. equorum Japan isolate (nearly complete), and the complete mtDNA sequences of P. univalens Switzerland and USA isolates. The complete mtDNA genome of P. equorum China isolate is 13,899 base pairs (bp), making it the smallest of the four genomes. All four Parascaris mt genomes are circular, and all genes are transcribed in the same direction. The P. equorum mtDNA genome consists of 12 protein-coding genes, two ribosomal RNA genes, 22 transfer (t) RNA genes and one non-coding region, which is consistent with P. equorum Japan isolate and P. univalens Switzerland isolate but distinct from P. univalens USA isolate, which has 20 tRNA genes. Differences in nucleotide sequences of the four entire mt genomes range from 0.1–0.9%, and differences in total amino acid sequences of protein-coding genes are 0.2–2.1%. Phylogenetic analyses showed that the four Parascaris species clustered in a clade, indicating that P. equorum and P. univalens are very closely related. These mt genome datasets provide genetic evidence that P. equorum and P. univalens may represent the same species, which will be of use in further studies of the taxonomy, systematics and population genetics of ascarids and other nematodes.

Steinernema khuongi n. sp. (Panagrolaimomorpha, Steinernematidae), a new entomopathogenic nematode species from Florida, USA

Abstract: In this study, molecular (ribosomal sequence data), morphological and cross-hybridization properties were used to identify a new Steinernema sp. from Florida, USA. Molecular and morphological data provided evidence for placing the novel species into Clade V, or the ‘glaseri-group’ of Steinernema spp. Within this clade, analysis of sequence data of the rDNA genes, 28S and internal transcribed spacer (ITS), depicted the novel species as a distinctive entity and closely related to S. glaseri and S. cubanum. Additionally, cross-hybridization assays showed that the new species is unable to interbreed with either of the latter two species, reinforcing its uniqueness from a biological species concept standpoint. Key morphological diagnostic characters for S. khuongi n. sp. include the mean morphometric features of the third-stage infective juveniles: total body length (average: 1066 μm), tail length (average: 65 μm), location of the excretory pore (average: 80.5 μm) and the values of c (average: 16.4), D% (average: 60.5), E% (average: 126) and H% (average: 46.6). Additionally, males can be differentiated from S. glaseri and S. cubanum by the values of several ratios: D% (average: 68), E% (average: 323) and SW% (average: 120). The natural distribution of this species in Florida encompasses both natural areas and citrus groves, primarily in shallow groundwater ecoregions designated as ‘flatwoods’. The morphological, molecular, phylogenetic and ecological data associated with this nematode support its identity as a new species in the S. glaseri-group.

In vitro nematicidal effect of Chenopodium ambrosioides and Castela tortuosa n-hexane extracts against Haemonchus contortus (Nematoda) and their anthelmintic effect in gerbils.

Abstract: The in vitro nematicidal effect of Chenopodium ambrosioides and Castela tortuosa n-hexane extracts (E-Cham and E-Cato, respectively) on Haemonchus contortus infective larvae (L3) and the anthelmintic effect of these extracts against the pre-adult stage of the parasite in gerbils were evaluated using both individual and combined extracts. The in vitro confrontation between larvae and extracts was performed in 24-well micro-titration plates. The results were considered 24 and 72 h post confrontation. The in vivo nematicidal effect was examined using gerbils as a study model. The extracts from the two assessed plants were obtained through maceration using n-hexane as an organic agent. Gerbils artificially infected with H. contortus L3 were treated intraperitoneally with the corresponding extract either individually or in combination. The results showed that the highest individual lethal in vitro effect (96.3%) was obtained with the E-Cham extract at 72 h post confrontation at 40 mg/ml, followed by E-Cato (78.9%) at 20 mg/ml after 72 h. The highest combined effect (98.7%) was obtained after 72 h at 40 mg/ml. The in vivo assay showed that the individual administration of the E-Cato and E-Cham extracts reduced the parasitic burden in gerbils by 27.1% and 45.8%, respectively. Furthermore, the anthelmintic efficacy increased to 57.3% when both extracts were administered in combination. The results of the present study show an important combined nematicidal effect of the two plant extracts assessed against L3 in gerbils.

Invasive slug Meghimatium pictum (Stoliczka, 1873) infected by Angiostrongylus costaricensis Morera & Céspedes, 1971, and the possible risk of human infection associated with grape consumption.

Abstract: Many molluscs may be infected with angiostrongylid larvae Following the histopathological diagnosis of abdominal angiostrongyliasis in a grape farmer from southern Brazil, molluscs in the area were investigated During a nocturnal search, 245 specimens of slugs were collected and identified as the invasive Chinese slug Meghimatium pictum Angiostrongylus costaricensis worms were recovered from mice that were experimentally infected with larvae obtained from 11 (45%) of the molluscs This study presents the first report of M pictum being identified as an intermediate host for A costaricensis Most of the slugs were collected from grape plants, which suggests that transmission may be associated with grape consumption

Clinical and haematological characteristics of human trichostrongyliasis.

Abstract: Trichostrongylus spp. are primarily parasites of ruminants, but humans can become infected as accidental hosts. Information about the clinical aspects of human trichostrongyliasis is limited. This study investigated the clinical and haematological characteristics of a large number of trichostrongyliasis patients. In the Fouman district of Guilan Province in northern Iran, during 2015-2016, 60 patients were identified as positive for Trichostrongylus spp., using stool examination methods. The clinical manifestations and demographic data of all patients were recorded and further analysed. Twenty-three patients (38.3%) were male and 37 (61.7%) were female. Among the individuals infected only with Trichostrongylus, only nine patients (16.4%) were asymptomatic. Forty-six patients (83.6%) presented with gastrointestinal (76.3%), pulmonary (30.9%) and cutaneous (12.7%) symptoms. No statistically significant relationship was found between clinical manifestations and sex or age groups. Ten patients (18.1%) revealed eosinophilia and five (9.1%) presented with hypochromic microcytic anaemia. The relationship between eosinophilia and age group, sex and clinical manifestations showed no statistical significance. Our study indicated that trichostrongyliasis may be a major parasitic aetiology for gastrointestinal symptoms and eosinophilia in rural residents of endemic areas.

Genotype characterization of livestock and human cystic echinococcosis in Mazandaran province, Iran.

Abstract: Echinococcus granulosus is a helminth from the family Taeniidae, which causes cystic echinococcosis (CE) in humans and diverse livestock around the world. The identification of existing genotypes in different regions is a major step towards the prevention and establishment of control programmes for the disease. This study aimed to detect CE genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer-1 (ITS1) gene and sequencing of the cytochrome c oxidase subunit 1 (Cox1) gene in isolates from the central part of Mazandaran province, northern Iran. Forty isolates were collected from sheep, 17 from cattle and 6 from human formalin-fixed paraffin-embedded tissues (FFPE). The ITS1 and Cox1 genes were successfully amplified by PCR in 41 and 42 samples, respectively. PCR-RFLP and sequencing showed that all isolates had the G1-G3 genotypes in this study. Out of 31 isolates subjected to sequencing for the Cox1 gene, 80.7% had the G1 genotype. G2 (16.1%) and G3 (3.2%) genotypes were observed in five sheep and one cattle samples, respectively. Five human isolates were also sequenced for the ITS1 gene, which showed that all samples belonged to the G1 genotype. Ten haplotypes were determined among the isolates by alignment analysis of the Cox1 gene. In summary, this study demonstrated that G1 was the dominant genotype circulating between humans and livestock in the studied region. Furthermore, high genotypic diversity among the CE isolates was observed.

Bulk tank milk ELISA to detect IgG1 prevalence and clustering to determine spatial distribution and risk factors of Fasciola hepatica-infected herds in Mexico.

Abstract: Fasciola hepatica is a helminth parasite that causes huge economic losses to the livestock industry worldwide Fasciolosis is an emerging foodborne zoonotic disease that affects both humans and grazing animals This study investigated the associations between climatic/environmental factors (derived from satellite data) and management factors affecting the spatial distribution of this liver fluke in cattle herds across different climate zones in three Mexican states A bulk-tank milk (BTM) IgG1 enzyme-linked immunosorbent assay (ELISA) test was used to detect F hepatica infection levels of 717 cattle herds between January and April 2015 Management data were collected from the farms by questionnaire The parasite's overall herd prevalence and mean optical density ratio (ODR) were 6276% and 067, respectively The presence of clustered F hepatica infections was studied using the spatial scan statistic Three marked clusters in the spatial distribution of the parasite were observed Logistic regression was used to test three models of potential statistical association from the ELISA results using climatic, environmental and management variables The final model based on climatic/environmental and management variables included the following factors: rainfall, elevation, proportion of grazed grass in the diet, contact with other herds, herd size, parasite control use and education level as significant predictors Geostatistical kriging was applied to generate a risk map for the presence of parasites in dairy herds in Mexico In conclusion, the spatial distribution of F hepatica in Mexican cattle herds is influenced by multifactorial effects and should be considered in developing regionally adapted control measures

Distribution of the newly invasive New Guinea flatworm Platydemus manokwari (Platyhelminthes: Geoplanidae) in Thailand and its potential role as a paratenic host carrying Angiostrongylus malaysiensis larvae

Abstract: Invasive species constitute one of the most serious threats to biodiversity and ecosystems, and they potentially cause economic problems and impact human health. The globally invasive New Guinea flatworm, Platydemus manokwari (Platyhelminthes: Geoplanidae), has been identified as a threat to terrestrial biodiversity, particularly soil-dwelling native species (e.g. molluscs, annelids and other land planarians), and is listed among 100 of the world's worst invasive alien species. We report here, for the first time, P. manokwari occurrences in many locations throughout Thailand, using voluntary digital public participation from the social network portals associated with the Thailand Biodiversity Conservation Group and collections of living flatworm specimens. Mitochondrial cytochrome c oxidase subunit I (COI) sequences confirmed that all collected flatworms were P. manokwari and placed them in the “world haplotype” clade alongside other previously reported specimens from France, Florida (USA), Puerto Rico, Singapore, French Polynesia, New Caledonia, and the Solomon Islands. In addition, infective stage larvae (L3) of the nematode Angiostrongylus malaysiensis were found in the flatworm specimens, with a 12.4% infection rate (15/121 specimens examined). Platydemus manokwari occurrence in Thailand and its capacity to carry L3 of Angiostrongylus should be of concern to biodiversity conservation and human health practitioners, because this invasive flatworm species may be involved in the life cycle of angiostrongylid worms in Thailand.

Limited parasite acquisition by non-native Lepomis gibbosus (Actinopterygii: Centrarchidae) at two ponds in the Upper Rhine basin, Germany.

Abstract: Metazoan parasite communities of Lepomis gibbosus (Centrarchidae), one of the most successfully introduced fish species in Europe, were studied at two isolated ponds (Knielingen, Tropfen) along the Upper Rhine in Germany. Nine parasite taxa were observed, including North American species co-introduced to Europe (ancyrocephalid monogeneans, diplostomid trematodes), circumpolar species infecting L. gibbosus in both their native and non-native ranges (bothriocephalid cestodes) and locally acquired parasitic nematodes. Both parasite communities consisted predominantly of North American species. Acquisition of local parasites was not observed at Tropfen, where the fish community comprised just two species, with L. gibbosus dominant. Low prevalence and abundance of acquired parasites was found at Knielingen, which supported a diverse fish community. At Tropfen, a high abundance of the North American parasite Posthodiplostomum centrarchi probably contributed to the lower condition index, hepatomegaly and splenomegaly observed. Due to low local parasite competency, L. gibbosus appears to have no significant impact on parasite dynamics in affected habitats.

Molecular characterization of Toxocara spp. eggs isolated from public parks and playgrounds in Shiraz, Iran

Abstract: Human toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

Records of opecoeline species Pseudopecoelus cf. vulgaris and Anomalotrema koiae Gibson & Bray, 1984 (Trematoda, Opecoelidae, Opecoelinae) from fish of the North Pacific, with notes on the phylogeny of the family Opecoelidae.

Abstract: Opecoelid species Pseudopecoelus cf. vulgaris and Anomalotrema koiae Gibson & Bray, 1984 were found in fish collected in the boreal waters of the North Pacific. Pseudopecoelus cf. vulgaris differs from Pseudopecoelus vulgaris (Manter, 1934) in terms of the egg size. This is the first record of A. koiae in the North Pacific, and the second of Pseudopecoelus cf. vulgaris. Previously, A. koiae was recorded only in North Atlantic fish. Partial sequences of 28S rDNA obtained for these two species and six other previously unsequenced representatives of the family Opecoelidae were analysed together with data from GenBank. Phylogenetic analysis supports the allocation of the six clades of opecoelids – Helicometrinae, Opecoelinae, Opistholebetinae, ‘freshwater Plagioporinae’, ‘marine Plagioporinae B’ and ‘marine Plagioporinae C’, and confirms the paraphyly of the group ‘deep-sea Plagioporinae’. Our phylogeny does not support previous hypotheses about the monophyly of opecoelines with a uroproct.

Antifilarial activity of azadirachtin fuelled through reactive oxygen species induced apoptosis: a thorough molecular study on Setaria cervi.

Abstract: Efficacious therapeutic strategies against lymphatic filariasis are always sought after However, natural products are a promising resource for developing effective antifilarial agents Azadirachtin, a significant tetranortriterpenoid phytocompound found in Azadirachta indica, was evaluated in vitro for antifilarial potential against the filarial parasite Setaria cervi Dye exclusion and MTT assay confirmed the antifilarial potential of azadirachtin against S cervi with a median lethal dose (LC50) of 628 μg/ml for microfilariae (mf), and 955 μg/ml for adult parasites Morphological aberrations were prominent in the histological sections of the azadirachtin-exposed parasites Moreover, alterations in the reactive oxygen species (ROS) parameters in treated parasites were evident Induction of apoptosis in treated parasites was confirmed by DNA laddering, acridine orange (AO)/ethidium bromide (EtBr) double staining and in situ DNA fragmentation The downregulation of anti-apoptotic CED-9 and upregulation of proapoptotic EGL-1, CED-4 and CED-3 at both the transcription and translation levels confirmed apoptosis execution at the molecular level Changes in the gene expressions of nuc-1, cps-6 and crn-1 further clarified the molecular cause of DNA degradation Furthermore, azadirachtin was found to be non-toxic in both in vitro and in vivo toxicity analyses Therefore, the experimental evidence detailed the pharmacological effectiveness of azadirachtin as a possible therapeutic agent against filariasis

Determinants of parasite distribution in Arctic charr populations: catchment structure versus dispersal potential.

Abstract: Parasite distribution patterns in lotic catchments are driven by the combined influences of unidirectional water flow and the mobility of the most mobile host. However, the importance of such drivers in catchments dominated by lentic habitats are poorly understood. We examined parasite populations of Arctic charr Salvelinus alpinus from a series of linear-connected lakes in northern Norway to assess the generality of lotic-derived catchment-scale parasite assemblage patterns. Our results demonstrated that the abundance of most parasite taxa increased from the upper to lower catchment. Allogenic taxa (piscivorous birds as final host) were present throughout the entire catchment, whereas their autogenic counterparts (charr as final hosts) demonstrated restricted distributions, thus supporting the theory that the mobility of the most mobile host determines taxa-specific parasite distribution patterns. Overall, catchment-wide parasite abundance and distribution patterns in this lentic-dominated system were in accordance with those reported for lotic systems. Additionally, our study highlighted that upper catchment regions may be inadequate reservoirs to facilitate recolonization of parasite communities in the event of downstream environmental perturbations.

Survey of trematodes in intertidal snails from Patagonia, Argentina: new larval forms and diversity assessment.

Abstract: Larval trematodes are the main parasites of snails, and they play a crucial role because they usually castrate their snail hosts and can thus alter their population and community dynamics. This study involved a survey of seven gastropod species (Crepipatella dilatata, Fissurella radiosa, Nacella magellanica, Pareuthria fuscata, Siphonaria lessonii, S. lateralis and Trophon geversianus) parasitized by 12 trematode species (one hemiurid, one gymnophallid, two lepocreadiids, two microphallids, one notocotylid, two renicolids, one philophtalmid, one schistosomatid and one zoogonid) from southern Patagonia (47°S, 65°W), Argentina. Only F. radiosa was free of parasites. The study included the description of five new larvae, based on morphological and molecular information, and a comparison of the parasite diversity with that of a northern locality (42°S, 64°W), characterized by a lower mollusc diversity. Species richness and diversity of parasites were higher in the southern site. This suggests a correlation between the level of parasitism and the diversity of molluscs (first intermediate hosts), which is higher at the high-latitude site and seems to attract shorebirds, which disperse the digenean eggs and facilitate the completion of their life cycles. These results support the notion that parasitism is influenced by large-scale factors such as biogeographical patterns, and small-scale factors such as diversity or abundance of intermediate and definitive hosts.

Immunohistochemical observation of local inflammatory cell infiltration in the host-tissue reaction site of human hydatid cysts.

Abstract: The aim of this study was to evaluate the pattern of local immune cell infiltration in human cystic echinococcosis (CE) by identifying the subtypes of immune cells using immunohistochemistry (IHC). Fifty surgically removed hydatid cyst samples and surrounding tissues were collected from patients referred to Al-Zahra Hospital, Isfahan, Iran. IHC was performed on the surrounding host tissue of hydatid cysts using anti-human CD3, CD19, CD8, CD4, CD68, CD56, Ki-67 and Foxp3 (forkhead box P3) antibodies. The results were then compared to hepatocellular carcinoma and chronic hepatitis. In the host-tissue reaction site of liver hydatid cysts, a distinct pattern of local immune cell response, which outwardly consisted of a pack of the fibrous elements, a layer of palisading macrophages, an eosinophil-containing layer and a layer of accumulated lymphocytes, was observed. However, in some cases there were no positive cells for CD56+ natural killer cells and Foxp3+ regulatory T cells. The CD3+ T cells were the predominant inflammatory cells in all groups, followed by CD19+ B cells. It can be concluded that different immune cells are involved in the local response to human hydatid cysts.