Transforming gastrointestinal helminth parasite identification in vertebrate hosts with metabarcoding: a systematic review.

BACKGROUND: Gastrointestinal helminths are a very widespread group of intestinal parasites that can cause major health issues in their hosts, including severe illness or death. Traditional methods of helminth parasite identification using microscopy are time-consuming and poor in terms of taxonomic resolution, and require skilled observers. DNA metabarcoding has emerged as a powerful alternative for assessing community composition in a variety of sample types over the last few decades. While metabarcoding approaches have been reviewed for use in other research areas, the use of metabarcoding for parasites has only recently become widespread. As such, there is a need to synthesize parasite metabarcoding methodology and highlight the considerations to be taken into account when developing a protocol. METHODS: We reviewed published literature that utilized DNA metabarcoding to identify gastrointestinal helminth parasites in vertebrate hosts. We extracted information from 62 peer-reviewed papers published between 2014 and 2023 and created a stepwise guide to the metabarcoding process. RESULTS: We found that studies in our review varied in technique and methodology, such as the sample type utilized, genetic marker regions targeted and bioinformatic databases used. The main limitations of metabarcoding are that parasite abundance data may not be reliably attained from sequence read numbers, metabarcoding data may not be representative of the species present in the host and the cost and bioinformatic expertise required to utilize this method may be prohibitive to some groups. CONCLUSIONS: Overall, using metabarcoding to assess gastrointestinal parasite communities is preferable to traditional methods, yielding higher taxonomic resolution, higher throughput and increased versatility due to its utility in any geographical location, with a variety of sample types, and with virtually any vertebrate host species. Additionally, metabarcoding has the potential for exciting new discoveries regarding host and parasite evolution.

An evolutionary molecular adaptation of an unusual stefin from the liver fluke Fasciola hepatica redefines the cystatin superfamily.

Fasciolosis is a worldwide parasitic disease of ruminants and an emerging human disease caused by the liver fluke Fasciola hepatica. The cystatin superfamily of cysteine protease inhibitors is composed of distinct families of intracellular stefins and secreted true cystatins. FhCyLS-2 from F. hepatica is an unusual member of the superfamily, where our sequence and 3D structure analyses in this study revealed that it combines characteristics of both families. The protein architecture demonstrates its relationship to stefins, but FhCyLS-2 also contains the secretion signal peptide and disulfide bridges typical of true cystatins. The secretion status was confirmed by detecting the presence of FhCyLS-2 in excretory/secretory products, supported by immunolocalization. Our high-resolution crystal structure of FhCyLS-2 showed a distinct disulfide bridging pattern and functional reactive center. We determined that FhCyLS-2 is a broad specificity inhibitor of cysteine cathepsins from both the host and F. hepatica, suggesting a dual role in the regulation of exogenous and endogenous proteolysis. Based on phylogenetic analysis that identified several FhCyLS-2 homologues in liver/intestinal foodborne flukes, we propose a new group within the cystatin superfamily called cystatin-like stefins.

Gastrointestinal helminths in brown trout (Salmo trutta Linnaeus, 1758) captured in Galician rivers (NW Spain).

Specimens of Salmo trutta (n = 613) captured by local anglers in different rivers in Galicia (NW Spain) during the 2015 fishing season (15 March-15 August) were examined. In total 1479 adult helminths were recovered from the gastrointestinal tracts of 221 fish. Moreover, the microscopic observation of the sediments obtained, previous diphasic concentration, revealed the presence of helminth eggs in 485 trout specimens. The following species were identified by morphological and molecular analysis: Crepidostomum metoecus (8.97%) (Trematoda); Salmonema ephemeridarum (16.97%), Raphidascaris acus (9.46%) and Pseudocapillaria sp. (2.12%) (Nematoda); and Echinorhynchus truttae (8.48%) (Acanthocephala). The prevalence, mean intensity and mean abundance of each helminth species were determined in relation to size/age of the fish. The helminth infracommunity comprised a maximum of four species and the species richness was S = 5. The biological cycles of most of the helminth species recovered are dependent on benthic macroinvertebrate fauna, which, in turn, is influenced by the water quality. Therefore, any changes that take place in the aquatic ecosystem (due to anthropogenic activities or climate change) may be reflected in the helminth composition.

Infection with the fox lungworm (Crenosoma vulpis) in two dogs from New England - Two clinical reports and updated geographic distribution in North America.

Crenosoma vulpis, the fox lungworm, is a helminth parasite endemic to the fox population of New England. Domestic dogs are susceptible to infection via ingestion of snails and slugs. Two dogs from New England were diagnosed with C. vulpis. The predominant clinical sign in both dogs was a chronic cough. Treatment with steroids and antibiotics only temporarily relieved clinical signs. Thoracic radiographs in both dogs revealed bronchial patterns. Endotracheal washes were performed in each dog revealing marked, mixed inflammation consisting mainly of neutrophils with eosinophils in lesser numbers. Helminth larvae could also be visualized on cytology. A fecal flotation revealed helminth larvae in one dog but failed to identify larvae in the second dog. The diagnosis of C. vulpis was confirmed via PCR analysis and sequencing of samples from both endotracheal washes. One dog was treated with fenbendazole (50 mg/kg PO q24h for 14 days), enrofloxacin (13 mg/kg PO q 24 h for 5 days), and a tapering protocol of prednisone (20 mg PO q12h for 5 days, 20 mg PO q24h for 5 days, then 20 mg PO q48h for 10 days). The second dog was treated with fenbendazole (50 mg/kg PO q24h for 10 days) with an additional 7 days of febantel and two doses of milbemycin, achieving complete resolution of clinical signs. This lungworm is becoming increasingly more prevalent in domestic dogs worldwide and may be more prevalent in New England than previously thought. Veterinary practitioners of New England should include this respiratory helminth as a differential in dogs with respiratory signs, and respiratory washes and Baermann fecal examinations are warranted in dogs presenting with non-specific respiratory clinical signs.

Role of l-arginine and CD11b+Gr-1+ cells in immunosuppression induced by Heligmosomoides polygyrus bakeri.

Myeloid-derived suppressor cells (MDSCs) are heterogeneous population of monocyte and granulocyte progenitors that are highly suppressive against T cells. In BALB/c mice infected with a nematode Heligmosomoides polygyrus bakeri, we studied the dynamics of MDSCs, identified as CD11b+Gr-1+, induction in different tissues along with the development of parasite infection. We observed that MDSC-like cells are induced both by larvae and adult stages of H polygyrus bakeri. Gr-1+ cells of suppressive phenotype are recruited in the bone marrow, peripheral blood and peritoneal cavity during histotropic phase of infection and are present at that time in the intestine wall, where worms reside. Later, during intestinal phase, suppressive Gr-1+ cells increased in mesenteric lymph nodes and the spleen. l-arginine metabolism was important for the protective immunity, and parasite-induced Gr-1+ cells showed elevated arginase-1 and iNOS expression. Inhibition of arginase-1 and l-arginine administration caused reduced level of infection that coincided with weaker suppressive phenotype of Gr-1+ cells. We identified that l-arginine pathway activation and induction of MDSC-like cells characterize immunosuppressive state during H polygyrus bakeri infection in mice. Our findings confirm the role of MDSCs in parasitic infections and point l-arginine pathway as a potential target for immunomodulation during nematode infections.

Integrative taxonomy of Sprentascaris mahnerti and phylogeny of raphidascaridid nematodes (Nematoda: Ascaridoidea: Raphidascarididae) suggest the monophyly of Sprentascaris.

Sprentascaris mahnerti (Nematoda: Raphidascarididae) collected from Loricariichthys labialis (Siluriformes: Loricariidae) in the Pantanal wetlands, State of Mato Grosso do Sul (Brazil), was redescribed using light and scanning electron microscopy (SEM), and genetically characterised along with two other raphidascaridids: Raphidascaroides brasiliensis and Ro. moraveci. Due to the systematic discussion regarding Raphidascaris and Sprentascaris, as well as the poor knowledge about the phylogenetic relationships within Raphidascarididae, phylogenies were reconstructed based on partial sequences of the 18S and 28S nuclear rRNA gene, the nuclear ITS1-5.8S-ITS2 and the cytochrome c oxidase subunit I (cox1) mtDNA. Morphological study of S. mahnerti, confirmed some previously described features, revealed new characteristics and permitted to elucidate some inconsistencies noted in the literature. Morphological and genetic characterisation of S. mahnerti supported its validity. Phylogenetic reconstructions supported the monophyly of Sprentascaris, which has three pairs of interlabial conspicuous cuticular projections as a synapomorphy. The relationships among several lineages of raphidascaridids were unsolved, albeit Goezia and Ichthyascaris formed well-supported monophyletic assemblages, in which the first included species with no relations regarding the habitat of hosts and the geographic origin. The present findings represent one more step towards the understanding of the interrelationships of raphidascaridid nematodes. In this sense, Sprentascaris should be considered valid as an independent lineage from Raphidascaris.

New species and new records of camallanid nematodes (Nematoda, Camallanidae) from marine fishes and sea snakes in New Caledonia.

Recent examinations of camallanid nematodes (Camallanidae) from marine fishes off New Caledonia, collected in the years 2003-2011, revealed the presence of the following five new species of Procamallanus Baylis, 1923, all belonging to the subgenus Spirocamallanus Olsen, 1952: Procamallanus (Spirocamallanus) dispar n. sp. from the common ponyfish Leiognathus equulus (type host) and the striped ponyfish Aurigequula fasciata (both Leiognathidae, Perciformes); Procamallanus (Spirocamallanus) bothi n. sp. from the leopard flounder Bothus pantherinus (Bothidae, Pleuronectiformes); Procamallanus (Spirocamallanus) hexophtalmatis n. sp. from the speckled sandperch Parapercis hexophtalma (Pinguipedidae, Perciformes); Procamallanus (Spirocamallanus) synodi n. sp. from the sand lizardfish Synodus dermatogenys (Synodontidae, Aulopiformes); and Procamallanus (Spirocamallanus) thalassomatis n. sp. from the yellow-brown wrasse Thalassoma lutescens (Labridae, Perciformes). These are described based on light and scanning electron microscopical (SEM) studies. An additional three congeneric nematodes unidentifiable to species are reported from perciform fishes and a shark: Procamallanus (Spirocamallanus) sp. 3 of Moravec et al., 2006, Procamallanus (Spirocamallanus) sp. 1, and Procamallanus (Spirocamallanus) sp. 2. Ten fish species are recorded as new hosts for Camallanus carangis Olsen, 1954. Two camallanids, Procamallanus (Spirocamallanus) sp. 3 (subgravid female) and Camallanus carangis (fourth-stage larva) were also found in the digestive tract of the New Caledonian sea krait Laticauda saintgironsi, serving apparently as postcyclic and paratenic hosts, respectively, for these fish nematodes.

TITLE: Nouvelles espèces et nouveaux signalements de nématodes camallanidés (Nematoda, Camallanidae) provenant de poissons marins et de serpents de mer en Nouvelle-Calédonie. ABSTRACT: L'étude récente de nématodes camallanidés (Camallanidae) de poissons marins capturés au large de la Nouvelle-Calédonie, recueillis de 2003 à 2011, a révélé la présence des cinq nouvelles espèces suivantes de Procamallanus Baylis, 1923, toutes appartenant au sous-genre Spirocamallanus Olsen, 1952 : Procamallanus (Spirocamallanus) dispar n. sp. de Leiognathus equulus (hôte-type) et de Aurigequula fasciata (tous deux Leiognathidae, Perciformes) ; Procamallanus (Spirocamallanus) bothi n. sp. de Bothus pantherinus (Bothidae, Pleuronectiformes) ; Procamallanus (Spirocamallanus) hexophtalmatis n. sp. de Parapercis hexophtalma (Pinguipedidae, Perciformes) ; Procamallanus (Spirocamallanus) synodi n. sp. de Synodus dermatogenys (Synodontidae, Aulopiformes) ; et Procamallanus (Spirocamallanus) thalassomatis n. sp. de Thalassoma lutescens (Labridae, Perciformes). Ces espèces sont décrites sur la base d'études réalisées au microscope électronique à balayage et au microscope photonique. Trois autres nématodes congénères non identifiables au niveau spécifique sont signalés chez des poissons perciformes et un requin : Procamallanus (Spirocamallanus) sp. 3 de Moravec et al., 2006, Procamallanus (Spirocamallanus) sp. 1 et Procamallanus (Spirocamallanus) sp. 2. Dix espèces de poissons sont rapportées comme nouveaux hôtes pour Camallanus carangis Olsen, 1954. Deux camallanidés, Procamallanus (Spirocamallanus) sp. 3 (femelles subgravides) et Camallanus carangis (larve du quatrième stade) ont également été trouvés dans le tube digestif du serpent marin de Nouvelle-Calédonie, Laticauda saintgironsi, qui semble servir respectivement d'hôte post-cyclique et d'hôte paraténique pour ces nématodes de poissons.

Anti-inflammatory Trained Immunity Mediated by Helminth Products Attenuates the Induction of T Cell-Mediated Autoimmune Disease.

Recent studies have suggested that the innate immune system can display characteristics of immunological memory and this has been called "innate immune memory" or "trained immunity." Certain fungal products have been shown to induce epigenetic imprinting on monocytes/macrophages that results in heightened inflammatory responses to subsequent stimuli. Here we report that innate immune cells can be trained to be more anti-inflammatory following exposure to products of a helminth pathogen. Macrophages trained in vitro with Fasciola hepatica total extract (FHTE) had enhanced IL-10 and IL-1RA, but reduced TNF production upon re-stimulation with FHTE or TLR ligands and this was reversed by inhibitors of DNA methylation. In contrast, macrophages trained with ß-glucan or Bacillus Calmette-Guérin had enhanced TNF production upon re-stimulation with Pam3cys or LPS. Furthermore, FHTE-trained macrophages had enhanced expression of markers of alternative activated macrophages (AAM). Macrophages from mice treated with FHTE expressed markers of AAM and had heightened IL-10 and IL-1RA production in response to FHTE or TLR ligands and had suppressed TNF and IL-12p40 production. Macrophages from mice treated with FHTE had reduced APC function and inhibited IL-17 production and the encephalitogenic activity of T cells in the experimental autoimmune encephalomyelitis (EAE) model. In addition, mice pre-treated with FHTE were resistant to induction of EAE and this was associated with a significant reduction in IL-17-producing γδ and CD4 T cells infiltrating the CNS. Our findings reveal that cells of the innate immune system can be trained in vitro or in vivo to be more anti-inflammatory by exposure to helminth products and this protects mice against the induction of a T cell-mediated autoimmune disease.

A systematic review of parasites and micropredators of non-avian reptiles (Reptilia=Sauropsida) in Chile.

Knowledge about the identity of parasites in vertebrates is relevant because of their influence on ecological processes and health of their hosts. This is particularly important for groups of animals currently facing conservation issues, such as reptiles. The diversity of species and supra-specific taxa of microparasites and macroparasites (such as helminths and arthropods) present in non-avian reptiles in Chile was analyzed through a systematic review. A total of 49 scientific documents (thesis projects, abstracts in congresses, book chapters and peer-reviewed articles) concerning parasites, taxonomic descriptions and parasite ecology were included in this review. The suborder Iguania was among the most studied group with 35 native species recognized as hosts of parasites and 39 species of parasites reported up to the category of species or genus. Liolaemus tenuis was the species with the most taxa of recorded parasites (11 taxa, and only three to species level), but Liolaemus chillanensis had the highest number of identified species of parasites. In addition, only one native species of Serpentes, one of Gymnophthalmoidea, and two of Gekkota, as well as some reports of exotic species, were recorded as hosts. Among parasites, Trombidiformes was the richest order with 10 species from the superfamily Pterygosomatoidea and 16 species from Trombiculoidea. Current knowledge about the richness of helminths is very limited and there were only a few records of microparasites. In general, there is an urgent need for the development of collaborative works between specialists in reptile taxonomy and epidemiology in parasitology destined to evaluate the consequences that reptiles and their parasites will suffer due to the ongoing processes of habitat loss, climate change and the still present taxonomic issues of the native reptiles.

100 Questions in Livestock Helminthology Research.

An elicitation exercise was conducted to collect and identify pressing questions concerning the study of helminths in livestock, to help guide research priorities. Questions were invited from the research community in an inclusive way. Of 385 questions submitted, 100 were chosen by online vote, with priority given to open questions in important areas that are specific enough to permit investigation within a focused project or programme of research. The final list of questions was divided into ten themes. We present the questions and set them briefly in the context of the current state of knowledge. Although subjective, the results provide a snapshot of current concerns and perceived priorities in the field of livestock helminthology, and we hope that they will stimulate ongoing or new research efforts.

Knowledge, attitude and practice with respect to soil contamination by Soil-Transmitted Helminths in Ibadan, Southwestern Nigeria.

Ancylostoma duodenale, Necator americanus, Ascaris lumbricoides and Trichuris trichiura, the four major Soil-transmitted helminths (STHs) and also Strongyloides stercoralis infect humans worldwide. Most studies on Soil-transmitted helminths (STHs) carried out in Ibadan were faecal examination based while relatively few soil examination based studies were carried out mostly around school premises using limited number of sampling locations. This study was therefore designed to assess the level of soil contamination with STH and associated risk factors in the eleven local government areas of Ibadan. A total of 1980 soil samples were purposively collected monthly, between January and December 2017, from toilet areas, refuse dump sites, house vicinities, playgrounds, roadsides/walk ways, and examined for the presence of parasite eggs or larvae using automated analysis of light microscopy images by computer. Well-structured questionnaires were administered to 620 consenting participants to obtain information on the Knowledge of STH infections, Attitudes and Practices towards the infections. Data were analyzed using SPSS version 21, Chi-square and ANOVA were used in the analysis at p < 0.05. Out of soil samples examined, 1087 (54.9%) had at least one species of parasite. The prevalence of hookworm was 74.5% followed by 50.2% and 37.2% for Strongyloides species larvae and adult respectively, and 25.1% for Ascaris species. The highest prevalence was observed in the refuse dump (74.2%) followed by toilet area (36.5%) while the lowest was at house vicinities (1.6%). Fifty-seven percent of the respondents use pit latrine while 20.6% still practice open defecation. A high transmission risk was observed as large percentage (66.8%) of the respondents showed inadequate knowledge of how to avoid STH infections. Moreover, 64.0% and 25.2% reported that they often walk barefooted and suck fingers respectively. The high prevalence of parasitic contamination of soil observed in the present study and the high proportion of respondent with inadequate knowledge of how to prevent transmission of STH pose a high risk of re-infections in the study area even after treatment. Therefore there is a need for proper education on parasite transmission in the area.

Pathological studies on helminth parasitic infection in mithun (Bos frontalis).

A pathological study was carried out in different helminth parasite infection organs in 110 animals during the period from 2010 to 2016. Histopathological examination of hydatid cysts in the liver, showed extensive fibrous connective tissue proliferation with eosinophilic and lymphocytic infiltration, in lung, parenchyma showed severe emphysema with infiltration of alveolar macrophages and lymphocytes infiltration. In Toxocara infection, polymorphonuclear infiltration in the sub-mucosa with hyper activity of goblet cells leading to occlusion of the gland was noted in the tissue sections. In amphistomiasis, intestinal mucosa showed diffuse hyperemia, thickening and ulceration. Petechiae were observed in the anterior part of the duodenum and jejunum. Presence of parasites in the abomasum revealed petechial haemorrhages on the mucousa. In Trichuris infection, the rectum showed hypertrophy, hyperplasia and hyperactivity of the goblet cells. In some places there is focal aggregation of mononuclear cells.

Two new species of nematode parasites, Cucullanus epinepheli sp. n. (Cucullanidae) and Procamallanus (Spirocamallanus) sinespinis sp. n. (Camallanidae), from marine serranid and haemulid fishes off New Caledonia.

Based on light and scanning electron microscopical studies, two new species of parasitic nematodes are described from marine perciform fishes off New Caledonia: Cucullanus epinepheli sp. n. (Cucullanidae) from the intestine of the brownspotted grouper Epinephelus chlorostigma (Valenciennes) (Serranidae) and Procamallanus (Spirocamallanus) sinespinis sp. n. from the intestine of the silver grunt Pomadasys argenteus (Forsskål) (Haemulidae). Cucullanus epinepheli sp. n. differs from its congeners mainly in possessing a unique structure of the anterior, elevated cloacal lip with a large posterior outgrowth covering the cloacal aperture and in the presence of cervical alae and two small preanal papillae on the median dome-shaped precloacal elevation. This is the second known nominal species of this genus parasitising fishes of the family Serranidae and the second representative of Cucullanus Müller, 1777 recorded from fishes in New Caledonian waters. Procamallanus (Spirocamallanus) sinespinis sp. n. is mainly characterised by 10-12 spiral ridges in the buccal capsule, the presence of wide caudal alae, three pairs of pedunculate preanal papillae, two unequally long spicules (465-525 µm and 218-231 µm) and by the tail tip with a knob-like structure in the male, and the broad, rounded tail with a terminal digit-like protrusion without cuticular spikes in the female. This is the fifth nominal species of the subgenus Spirocamallanus Olsen, 1952 reported from fishes in New Caledonian waters.

Condensed Tannins in the Gastrointestinal Tract of Cattle after Sainfoin (Onobrychis viciifolia) Intake and Their Possible Relationship with Anthelmintic Effects.

Condensed tannins' (CTs) fate along the digestive tract of ruminants may account for the variable efficacy of CTs against gastrointestinal nematodes. We analyzed CTs in the digesta of cattle fed sainfoin. With the acetone-butanol-HCl assay, the total CTs concentrations in the digesta were close to those in the diets (6.3 and 1.5% of DM in experiments 1 and 2, respectively); thus, CTs remained potentially largely undegraded/unabsorbed. With the thiolysis assay, CTs concentration was much higher in the abomasum (2.3% of DM; expt 1) compared with the rumen and intestines, along with higher mean size and prodelphinidins percentage, corroborating CTs efficacy reported only against Ostertagia ostertagi in the abomasum. In expt 2, the dietary levels of CTs were probably too low to demonstrate anthelmintic effects in the rumen. Overall, the level of CTs accessible to thiolysis is favored under the acidic conditions of the abomasum, which seems critical for anthelmintic activity.

Reduced helminth parasitism in the introduced bank vole (Myodes glareolus): More parasites lost than gained.

Introduced species are often less parasitised compared to their native counterparts and to ecologically similar hosts in the new environment. Reduced parasitism may come about due to both the loss of original parasites and low acquisition of novel parasites. In this study we investigated the intestinal helminth parasites of the introduced bank vole (Myodes glareolus) in Ireland. Results were compared to data from other European studies and to the intestinal helminth fauna of an ecologically similar native rodent in Ireland, the wood mouse (Apodemus sylvaticus). The helminth fauna of introduced bank voles exhibited low diversity with only 3 species recovered: Aspiculuris tianjinensis; Aonchotheca murissylvatici and Taenia martis larvae. In particular, no adult parasites with indirect life-cycles were found in bank voles suggesting that indirectly transmitted parasites are less likely to establish in invasive hosts. Also, the results of this study add support to the enemy release hypothesis.

Effect of Intestinal Tapeworm Clestobothrium crassiceps on Concentrations of Toxic Elements and Selenium in European Hake Merluccius merluccius from the Gulf of Lion (Northwestern Mediterranean Sea).

The capacity for heavy metal bioaccumulation by some fish parasites has been demonstrated, and their contribution to decreasing metal concentrations in tissues of parasitized fish has been hypothesized. The present study evaluated the effect of the cestode Clestobothrium crassiceps on the accumulation of trace elements in 30 European hake, Merluccius merluccius, in Spain (half of them infested by C. crassiceps). Tissue samples from all M. merluccius and specimens of C. crassiceps from the infected hakes were collected and stored until element analysis by inductively coupled plasma mass spectrometry (ICP-MS). Arsenic, mercury, and selenium were generally present in lower levels in the cestode than in all hake tissues. The mean value of the muscular Se:Hg molar ratio in the infested subsample was higher than that in hakes without cestodes. Values indicate that the edible part of infested hakes presents a lower amount of Cd and Pb in relation to noninfested hakes.

A new species of Paracreptotrema (Digenea, Plagiorchiformes, Allocreadiidae) infecting two species of poeciliids in Río Malila of the Río Pánuco basin, Hidalgo, México, with a key to the species of the genus.

Paracreptotremarosenthali sp. n. was discovered in the intestine of Xiphophorusmalinche and Pseudoxiphophorusjonesii, collected from the headwaters of Río Malila, tributary of Río Conzintla, in the Río Pánuco basin, Hidalgo, México, during 2008-2009. The new species differs from the five known species of Paracreptotrema Choudhury, Pérez-Ponce de León, Brooks & Daverdin, 2006 by having vitelline follicles that extend from a level anterior to the pharynx to mid-testes, the seminal vesicle which is more extensively folded, and a wider cirrus sac. The new species resembles Paracreptotremaheterandriae in the length of its ceca, which surpasses the posterior margin of the ovary but do not reach the testes. A key to the species of Paracreptotrema is provided.

Trichostrongylus colubriformis induces IgE-independent CD13, CD164 and CD203c mediated activation of basophils in an in vitro intestinal epithelial cell co-culture model.

Gastrointestinal nematodes pose a major risk to the farming of small ruminants worldwide. Infections are typically controlled by anthelmintics, however as resistance to anthelmintics increases, it is necessary that the mechanism of host responses are understood in order to develop alternative control options. It is hypothesised that basophils are involved in the initiation of an anti-parasite immune response, independent of IgE. In this study, the in vitro activation states of CD203c(+) basophil-like KU812 cells were determined in the presence of Trichostrongylus colubriformis parasitised HT29 epithelial cells with or without mucin. Cell surface expression of CD164, CD107a and CD13 antigens on gated CD203(+) cells were determined and qRT-PCR was used to examine gene expression changes of IL33 (a Th2 cytokine) and the high affinity IgE receptor (FcɛRIα) within the co-culture. When KU812 basophils encountered T. colubriformis and/or mucin in a parasitised epithelium, the basophils increased cell surface expression of CD13 and CD164 antigens, independent of IgE. T. colubriformis also increased the number of CD203c(+) KU812 cells that expressed CD13 and CD164 antigens. These data support the in vivo observations of T. colubriformis primary infections in guinea pigs and sheep.

Helminth Parasites of Eastern European Hedgehog (Erinaceus concolor) in Northern Iran.

BACKGROUND: Recently there is a high tendency among exotic pet owners for keeping hedgehogs. This mammal can transfer some significant zoonotic pathogens to human. Hence, the present study was conducted for the first time to prepare a list of helminth parasites of hedgehogs (Erinaceus concolor) in North of Iran. METHODS: Ten (four males and six females) road killed hedgehogs were collected during April to January 2011 in rural areas of Babol city, Mazandaran province, Iran. All of internal organs were scrutinized for helminth burden. The extracted specimens were fixed and preserved in 70% ethanol and then cleared in Lacto-phenol solution. Helminth identification was carried out according to available systematic keys. RESULTS: All the examined hedgehogs (100%) were infected with parasitic helminth as following: two hedgehogs (20%) were infected with Crenosoma striatum, four hedgehogs (40%) harbored Physaloptera clausa, one (10%) host had Hymenolepis erinacei and three (30%) of them were infected with Nephridiacanthus major. CONCLUSION: This is noteworthy that the current survey is the first report of helminth parasites fauna of Eastern European Hedgehog in Iran. Since, this is the first such investigation in our country, more researches are required to perform on unexplored areas of Iran in order to increase our knowledge regarding hedgehog parasitic diseases.