A new ring nematode, Xenocriconemella costaricense sp. nov., (Nematoda: Criconematidae) from Costa Rica.

During nematode surveys of natural vegetation in forests of La Cima de Copey de Dota, San José, San José province, Costa Rica, a Xenocriconemella species closely resembling X. macrodora and related species was found. Integrative taxonomical approaches demonstrated that it is a new species described herein as X. costaricense sp. nov. The new species is parthenogenetic (only females have been detected) and characterised by a short body (276-404 µm); lip region with two annuli, not offset, not separated from body contour; first lip annulus partially covering the second lip annulus. Stylet thin, very long (113-133 µm) and flexible, occupying 30.5-47.8% of body length. Excretory pore located from one or two annuli anterior to one or two annuli posterior to level of stylet knobs, at 42 (37-45) µm from anterior end. Female genital tract monodelphic, prodelphic, outstretched, and occupying 35-45% of body length, with vagina slightly ventrally curved (14-18 µm long). Anus located 6-11 annuli from the tail terminus. Tail conoid and bluntly rounded terminus, the last 2-3 annuli oriented dorsally. Results of molecular characterisation and phylogenetic analyses of D2-D3 expansion segments of 28S rRNA, ITS, and partial 18S rRNA, as well as cytochrome oxidase c subunit 1 gene sequences further characterised the new species and clearly separated it from X. macrodora and other related species (X. iberica, X. paraiberica, and X. pradense).

Three new species of free-living marine nematodes of the Microlaimus genus (Nematoda: Microlaimidae) from the continental shelf off northeastern Brazil (Atlantic Ocean).

Three new species of the Microlaimus genus (Nematoda: Microlaimidae) are described from sample sediments collected in the South Atlantic, along the Continental Shelf break of Northeastern Brazil. Microlaimus paraundulatus sp. n. possesses four setiform cephalic sensillae, a buccal cavity with three small teeth, arched and slender spicules and a wave-shaped gubernaculum. Microlaimus modestus sp. n. is characterized by four small cephalic sensillae, a buccal cavity with three teeth (one large dorsal tooth), cephalated spicules and a strongly arched gubernaculum in the distal region. Microlaimus nordestinus sp. n. is characterized by the following set of features: relatively long body, eight rows of hypodermal glands that extend longitudinally along the body and a funnel-shaped gubernaculum surrounding the spicules at the distal end. An amendment of the diagnosis is proposed for the genus.

Biomarkers of gastrointestinal nematodes in beef cattle raised in a tropical area.

Biomarkers are specific molecular, histological, or physiological characteristics of normal or pathogenic biological processes and are promising in the diagnosis of gastrointestinal nematodes (GINs). Although some biomarkers have been validated for infection by Ostertagia sp. in cattle raised in temperate regions, there is a lack of information for tropical regions. The aim of this project was to assess potential biomarkers and validate the most promising. In the first study, 36 bovines (Nelore breed) naturally infected by GINs were distributed into two groups: infected (not treated with anthelmintic) and treated (treated with fenbendazole on days 0, 7, 14, 21, 28, 42, and 56). The variables of interest were live weight, fecal egg count, hemogram, serum biochemical markers, phosphorus, gastrin, and pepsinogen. In the second step, pepsinogen was assessed in cattle of the Nelore breed distributed among three groups: infected (not treated with anthelmintic), MOX (treated with moxidectin), and IVM + BZD (treated with ivermectin + albendazole). In the first study, no difference between groups was found for weight, albumin, hematocrit (corpuscular volume [CV]), erythrocytes, or hemoglobin. Negative correlations were found between pepsinogen and both CV and albumin, and albumin was negatively correlated with the percentage of Haemonchus sp. in the fecal culture. Among the biomarkers, only pepsinogen differentiated treated and infected (beginning with the 28th day of the study). In the second study, a reduction in pepsinogen was found after anthelmintic treatment. Therefore, pepsinogen is a promising biomarker of worms in cattle naturally infected by the genera Haemonchus and Cooperia in tropical areas.

Endoparasitic helminth fauna and diet of Geophagus sveni (Pisces) in Upper Paraná River basin.

In freshwater ecosystems, parasite infection patterns are influenced by factors including spatial-temporal variations, host diet, and habitat. Fish often change diets, affecting their parasite communities. This study focused on non-native host fish Geophagus sveni, aiming to characterize diet and endoparasitic helminth fauna patterns in the invaded area, investigating spatial and seasonal possible differences of endoparasite infections and correlating with host diet, in São José dos Dourados River and Tietê River areas. The host fish were collected in these areas during the dry and rainy season using gillnets. The endoparasites were collected and preserved in alcohol and identified using taxonomic methods, and stomach contents were examined for diet analysis. Parasitism descriptors were calculated and evaluated spatially and seasonally by ANOVA and the Kruskal-Wallis tests. PERMANOVA assessed G. sveni diet differences, and RDA correlated the endohelminth abundance with the host diet. Two endoparasites were recorded: metacercariae of Austrodiplostomum compactum (Trematoda) and larvae and adults of Raphidascaris (Sprentascaris) lanfrediae (Nematoda). Spatial differences were observed for the mean abundance and prevalence of R. (S.) lanfrediae and A. compactum prevalence. Seasonal variations of parasitic descriptors occurred for the nematode in the Tietê River area. The detritus and aquatic insects were the most consumed items by G. sveni. Detritus consumption positively correlates with nematode abundance. The findings indicate that factors such as artificial channels and rainfall, which can influence resource availability, may affect the fish's diet and potentially influence the structure of its endoparasite community. The study emphasizes the importance of understanding trophic chain-transmitted parasites and calls for further research in Neotropical environments.

Description of Pudica wandiquei n sp (Heligmonellidae: Pudicinae), a nematode found infecting Proechimys simonsi (Rodentia: Echimyidae) in the Brazilian Amazon / Descrição de Pudica wandiquei n sp (Heligmonellidae: Pudicinae), nematódeo encontrado infectando Proechimys simonsi (Rodentia: Echimyidae) na Amazônia brasileira

A new species of nematode parasite of the subfamily Pudicinae (Heligmosomoidea: Heligmonellidae) is described from the small intestine of Proechimys simonsi (Rodentia: Echimyidae) from the locality of Nova Cintra in the municpality of Rodrigues Alves, Acre state, Brazil. The genus Pudica includes 15 species parasites of Neotropical rodents of the families Caviidae, Ctenomyidae, Dasyproctidae, Echimyidae, Erethizontidae, and Myocastoridae. Four species of this nematode were found parasitizing three different species rodents of the genus Proechimys in the Amazon biome. Pudica wandiquei n. sp. can be differentiated from all other Pudica species by the distance between the ends of rays 6 and 8 and the 1-3-1 pattern of the caudal bursa in both lobes.

Uma nova espécie de nematódeo da subfamília Pudicinae (Heligmosomoidea: Heligmonellidae) é descrito parasitando o intestino delgado de Proechimys simonsi (Rodentia: Echimyidae) em Nova Cintra, município de Rodrigues Alves, Estado do Acre, Brasil. O gênero Pudica inclui 15 espécies parasitas de roedores Neotropicais das famílias: Caviidae, Ctenomyidae, Dasyproctidae, Echimyidae, Erethizontidae e Myocastoridae. Destas, quatro espécies de nematódeos foram encontradas parasitando três diferentes espécies do roedor Proechimys no bioma Amazônia. Pudica wandiquei n. sp. é diferente das outras espécies de Pudica pela distância entre os raios 6 e 8 e pelo tipo da bolsa caudal, que é 1-3-1 em ambos os lados.

Nematodes and trematodes associated with terrestrial gastropods in Nottingham, England.

A parasitological survey of terrestrial slugs and snails was conducted at popular dog walking locations across the city of Nottingham, with the intensions of finding gastropods infected with parasites of medical (or veterinary) importance such as lungworm (metastrongyloid nematodes) and trematodes. A total of 800 gastropods were collected from 16 sites over a 225 km2 area. The extracted nematodes and trematodes were identified by molecular barcoding. Of the 800 gastropods collected, 227 were infected (172 had nematode infections, 37 had trematode infections and 18 had both nematode and trematode infections). Of the nematode infected gastropods genotyped, seven species were identified, Agfa flexilis, Angiostoma gandavense, Angiostoma margaretae, Cosmocerca longicauda, Phasmarhabditis hermaphrodita, Phasmarhabditis neopapillosa and an unknown Cosmocercidae species. Of the trematode infected gastropods genotyped, four species were identified, Brachylaima arcuate, Brachylaima fuscata, Brachylaima mesostoma and an unknown Plagiorchioidea species. No lungworm species were found within the city of Nottingham. To our knowledge, this study represents the first survey of gastropod-associated nematodes and trematodes in the East midlands of the United Kingdom.

Gastrointestinal helminth parasites of the threatened Australasian crested grebe (Podiceps cristatus australis, Gould 1844) in New Zealand, with descriptions of Baruscapillaria kamanae n. sp. (Nematoda: Trichuridae) and Cryptocotyle micromorpha n. sp. (Trematoda: Opisthorchiidae).

The Australasian crested grebe Podiceps cristatus australis, Gould 1844 is restricted to Australia and New Zealand, where it is listed as Threatened and Nationally Vulnerable. For the first time in New Zealand, we report on the parasitic helminths infecting three individuals from Lake Wanaka, Otago, using morphological and molecular tools. Seven helminth species were found in the gastrointestinal tract: 2 nematodes (Contracaecum ovale and Baruscapillaria kamanae n. sp.), 4 trematodes (Australapatemon minor, Cryptocotyle micromorpha n. sp., Tylodelphys darbyi and Neopetasiger neocomensis), and 1 cestode (Confluaria pseudofurcifera). Except for T. darbyi, all are new records for New Zealand. A change of orthography is proposed for Neopetasiger neocomensis and N. pseudoneocomensis. Cryptocotyle micromorpha n. sp. (Opisthorchiidae) is distinguished from similar species by its small size, wholly extracaecal vitellaria and anteriorly looped uterus. Baruscapillaria kamanae n. sp. (Trichuridae) is distinguished from other freshwater species by a combination of vulva and spicule morphology. The helminth parasites found here are mostly the same as those from the grebe in the northern hemisphere, indicating that they have been carried with the host species in its spread to Australasia. However, the parasite fauna may be depauperate due to a diminishing reservoir of intermediate hosts in that geographical migration.

Nematode community structure along elevation gradient in high altitude vegetation cover of Gangotri National Park (Uttarakhand), India.

The study was undertaken to understand the dynamics of soil nematode communities association along elevation gradient and associated variables in high-altitude regions of Western Himalaya. The diversity and distribution pattern of soil nematodes were investigated at four elevation ranges (Elv1, Elv2, Elv3, and Elv4 each of 500 m class) along altitudinal gradient (3000-5000 m). The nematode community comprised 58 genera of which 37 genera showed sensitivity towards altitudinal variation and the community structure also varied among elevation classes. It was found that elevation significantly affect the nematodes trophic group, diversity composition, and the nematode indices. Nematode generic richness and evenness index indicated a relatively low biodiversity of soil nematodes is supported at higher altitudes. Moreover, maturity indices reflected lower sustenance of k-strategic nematodes at higher elevations. Structure index depicted fewer connections in the soil food web at higher elevations. Nematode faunal profile showed low nutrient and low disturbance in the region. Carbon footprint of the whole nematode assemblage declined along the elevation. Overall substantial differences in the nematode composition, abundance, trophic structure, and contribution to belowground carbon cycling were observed with change in elevation. These findings could be utilized as useful tool in the long-term monitoring and to understand the region's soil health.

Pan-phylum genome-wide identification of sodium calcium exchangers reveal heterogeneous expansions and possible roles in nematode parasitism.

Calcium signaling is ubiquitous in nematode development from fertilization to cell specification to apoptosis. Calcium also regulates dauer entry in Caenorhabditis elegans, which corresponds to the infective stage of parasitic nematodes. In diverse parasites such as Trypanosoma cruzi and Toxoplasma gondii calcium has been shown to regulate host cell entry and egress, and perturbing calcium signaling represents a possible route to inhibit infection and parasitism in these species. Sodium calcium exchangers are considered the most important mechanism of calcium efflux, and our lab has previously characterized the sodium calcium exchanger gene family in C. elegans and studied the diversity of this family across a subset of specific nematode species. Here we build upon these data and explore sodium calcium exchangers across 108 species of nematodes. Our data reveal substantial differences in sodium calcium exchanger counts across the Phylum and detail expansions and contractions of specific exchanger subtypes within certain nematode clades. Finally, we also provide evidence for a role of sodium calcium exchangers in parasite activation by examining differentially expressed genes in non-activated versus activated infective stage larvae. Taken together our findings paint a heterogeneous picture of sodium calcium exchanger evolution across the Phylum Nematoda that may reflect unique adaptations to free-living and parasitic lifestyles.

PHI-base in 2022: a multi-species phenotype database for Pathogen-Host Interactions.

Since 2005, the Pathogen-Host Interactions Database (PHI-base) has manually curated experimentally verified pathogenicity, virulence and effector genes from fungal, bacterial and protist pathogens, which infect animal, plant, fish, insect and/or fungal hosts. PHI-base (www.phi-base.org) is devoted to the identification and presentation of phenotype information on pathogenicity and effector genes and their host interactions. Specific gene alterations that did not alter the in host interaction phenotype are also presented. PHI-base is invaluable for comparative analyses and for the discovery of candidate targets in medically and agronomically important species for intervention. Version 4.12 (September 2021) contains 4387 references, and provides information on 8411 genes from 279 pathogens, tested on 228 hosts in 18, 190 interactions. This provides a 24% increase in gene content since Version 4.8 (September 2019). Bacterial and fungal pathogens represent the majority of the interaction data, with a 54:46 split of entries, whilst protists, protozoa, nematodes and insects represent 3.6% of entries. Host species consist of approximately 54% plants and 46% others of medical, veterinary and/or environmental importance. PHI-base data is disseminated to UniProtKB, FungiDB and Ensembl Genomes. PHI-base will migrate to a new gene-centric version (version 5.0) in early 2022. This major development is briefly described.

VEuPathDB: the eukaryotic pathogen, vector and host bioinformatics resource center.

The Eukaryotic Pathogen, Vector and Host Informatics Resource (VEuPathDB, https://veupathdb.org) represents the 2019 merger of VectorBase with the EuPathDB projects. As a Bioinformatics Resource Center funded by the National Institutes of Health, with additional support from the Welllcome Trust, VEuPathDB supports >500 organisms comprising invertebrate vectors, eukaryotic pathogens (protists and fungi) and relevant free-living or non-pathogenic species or hosts. Designed to empower researchers with access to Omics data and bioinformatic analyses, VEuPathDB projects integrate >1700 pre-analysed datasets (and associated metadata) with advanced search capabilities, visualizations, and analysis tools in a graphic interface. Diverse data types are analysed with standardized workflows including an in-house OrthoMCL algorithm for predicting orthology. Comparisons are easily made across datasets, data types and organisms in this unique data mining platform. A new site-wide search facilitates access for both experienced and novice users. Upgraded infrastructure and workflows support numerous updates to the web interface, tools, searches and strategies, and Galaxy workspace where users can privately analyse their own data. Forthcoming upgrades include cloud-ready application architecture, expanded support for the Galaxy workspace, tools for interrogating host-pathogen interactions, and improved interactions with affiliated databases (ClinEpiDB, MicrobiomeDB) and other scientific resources, and increased interoperability with the Bacterial & Viral BRC.

Survey of nematodes associated with sugarcane in the state of Paraná, Brazil

Sugarcane-associated nematodes (Saccharum spp.) can reduce productivity up to 50%. Through the survey, it was possible to identify the main nematodes that occur in a certain region as a tool for designing the best management and control strategies. The aim of this study was to characterize the population of nematodes associated with sugarcane in the North Central, North Pioneiro and Northwest mesoregion of the state of Paraná, Brazil, quantify the nematode genera associated with the crop and identify the species of Pratylenchus and Meloidogyne. A total amount of 89 soil and root composite samples were collected in nine municipalities. Nematodes were extracted and counted in a Peters counting chamber under an optical light microscope. Morphological description followed identification keys. Pratylenchus spp. were identified by morphological characteristics; Meloidogyne spp. were identified by morphological characteristics and isoenzyme electrophoresis. Twelve genera of nematodes associated with sugarcane were identified: Pratylenchus, Meloidogyne, Helicotylenchus, Xiphinema, Mesocriconema, Trichodorus, Aphelenchus, Hoplolaimus, Tylenchus, Tylenchorhynchus, Ditylenchus, and Paratrichodorus. The genera Pratylenchus and Meloidogyne were found with the highest frequencies in the roots. Among the species of Pratylenchus, P. zeae and P. brachyurus were found, with P. zeae being the most frequent. Among the Meloidogyne species, only M. javanica was found. These results are essential to aid decision making in the management of phytonematodes, mainly in the development of new control strategies and in directing genetic breeding programs for development of sugarcane cultivars for the Paraná state.

A new species of Huffmanela (Nematoda: Trichosomoididae) parasitizing Colomesus psittacus (Tetraodontiformes) from Marajó Island, Pará State, Brazil / Uma nova espécie de Huffmanela (Nematoda: Trichosomoididae) parasitando Colomesus psittacus (Tetraodontiformes) da Ilha de Marajó, Pará, Brasil

Abstract The puffer fish Colomesus psittacus, is not commercialized on Marajó Island. They are captured as bycacth and discarded dead in the environment in artisanal fisheries that occur in the estuaries of northern Brazil. In this sense, the objective was to identify the parasites present in the gills and to evaluate the histopathological alterations caused by these nematodes of the genus Huffmanela. Fifty-five fish were analyzed, and thirty-five specimens showed the parasite in the gills. Morphological characteristics suggest that it is a new species of the genus Huffmanela, and the histopathological exams showed an edematous inflammation in the secondary lamella and the presence of eggs of this nematode, which is the first record of this parasite in C. psittacus in Brazil.

Resumo O baiacu amazônico Colomesus psittacus é um peixe de importância comercial para a aquariofilia, além de ser capturado como bycacth e descartado morto no ambiente nas pescarias artesanais que ocorrem nos estuários da região Norte do Brasil. Nesse sentido, objetivou-se identificar e descrever nematódeos do gênero Huffmanela em baiacus oriundos da ilha de Marajó, bem como as alterações histopatológicas causadas por eles. Cinquenta e cinco peixes foram analisados, e trinta e cinco espécimes apresentaram o parasita nas branquias. Características morfológicas sugerem tratar-se de uma nova espécie do gênero Huffmanela, e os exames histopatológicos demonstraram uma inflamação edematosa na lamela secundária e presença de ovos desse nematódeo. Esse fato se torna o primeiro registro desse gênero parasito em C. psittacus no Brasil.

Metabarcoding in two isolated populations of wild roe deer (Capreolus capreolus) reveals variation in gastrointestinal nematode community composition between regions and among age classes.

BACKGROUND: Gastrointestinal nematodes are ubiquitous for both domestic and wild ungulates and have varying consequences for health and fitness. They exist as complex communities of multiple co-infecting species, and we have a limited understanding of how these communities vary in different hosts, regions and circumstances or of how this affects their impacts. METHODS: We have undertaken ITS2 rDNA nemabiome metabarcoding with next-generation sequencing on populations of nematode larvae isolated from 149 fecal samples of roe deer of different sex and age classes in the two isolated populations of Chizé and Trois Fontaines in France not co-grazing with any domestic ungulate species. RESULTS: We identified 100 amplified sequence variants (ASVs) that were assigned to 14 gastrointestinal nematode taxa overall at either genus (29%) or species (71%) level. These taxa were dominated by parasites classically found in cervids-e.g. Ostertagia leptospicularis, Spiculopteragia spp. Higher parasite species diversity was present in the Trois Fontaines population than in the Chizé population including the presence of species more typically seen in domestic livestock (Haemonchus contortus, Bunostomum sp., Cooperia punctata, Teladorsagia circumcincta). No differences in parasite species diversity or community composition were seen in the samples collected from three zones of differing habitat quality within the Chizé study area. Young roe deer hosted the highest diversity of gastrointestinal nematodes, with more pronounced effects of age apparent in Trois Fontaines. The effect of host age differed between gastrointestinal nematode species, e.g. there was little effect on O. leptospicularis but a large effect on Trichostrongylus spp. No effect of host sex was detected in either site. CONCLUSIONS: The presence of some livestock parasite species in the Trois Fontaines roe deer population was unexpected given the isolation of this population away from grazing domestic livestock since decades. Overall, our results illustrate the influence of host traits and the local environment on roe deer nemabiome and demonstrate the power of the nemabiome metabarcoding approach to elucidate the composition of gastrointestinal nematode communities in wildlife.

Use of universal primers for the 18S ribosomal RNA gene and whole soil DNAs to reveal the taxonomic structures of soil nematodes by high-throughput amplicon sequencing.

Nematodes are abundant metazoans that play crucial roles in nutrient recycle in the pedosphere. Although high-throughput amplicon sequencing is a powerful tool for the taxonomic profiling of soil nematodes, polymerase chain reaction (PCR) primers for amplification of the 18S ribosomal RNA (SSU) gene and preparation of template DNAs have not been sufficiently evaluated. We investigated nematode community structure in copse soil using four nematode-specific (regions 1-4) and two universal (regions U1 and U2) primer sets for the SSU gene regions with two DNAs prepared from copse-derived mixed nematodes and whole soil. The major nematode-derived sequence variants (SVs) identified in each region was detected in both template DNAs. Order level taxonomy and feeding type of identified nematode-derived SVs were distantly related between the two DNA preparations, and the region U2 was closely related to region 4 in the non-metric multidimensional scaling (NMDS) based on Bray-Curtis dissimilarity. Thus, the universal primers for region U2 could be used to analyze soil nematode communities. We further applied this method to analyze the nematodes living in two sampling sites of a sweet potato-cultivated field, where the plants were differently growing. The structure of nematode-derived SVs from the two sites was distantly related in the principal coordinate analysis (PCoA) with weighted unifrac distances, suggesting their distinct soil environments. The resultant ecophysiological status of the nematode communities in the copse and field on the basis of feeding behavior and maturity indices was fairly consistent with those of the copse- and the cultivated house garden-derived nematodes in prior studies. These findings will be useful for the DNA metabarcoding of soil eukaryotes, including nematodes, using soil DNAs.

NEW RECORDS OF NEMATODES IN THE YELLOW-BILLED CUCKOO COCCYZUS AMERICANUS (CUCULIFORMES: CUCULIDAE) BASED ON A MIGRANT SPECIMEN FOUND IN BRAZIL.

The yellow-billed cuckoo, Coccyzus americanus, is a threatened passerine bird native to North America that migrates to overwinter in South America. Although migratory birds have drawn broad attention, given their assumed role in the long-distance dispersal of parasites, studies on the helminth fauna of this cuculid are scarce. In the present study, nematodes found in the gastrointestinal tract of a specimen of C. americanus found in Belo Horizonte, State of Minas Gerais, southeastern Brazil, were characterized morphologically. Five species of gastrointestinal nematodes belonging to 4 families were identified: Synhimantus (Dispharynx) nasuta (Rudolphi, 1819) and Synhimantus (Dispharynx) resticulaCanavan, 1929 (Acuariidae), Microtetrameres sp. (Tetrameridae), and Cyrnea piayaeSandground, 1929 (Habronematidae) and Subulura halli Barreto, 1918 (Subuluridae). Except for Microtetrameres sp., all other nematodes are reported in this host for the first time. Although it is difficult to accurately determine the geographical origin of infections, it is important to note that Sy. (D.) resticula and Su. halli are reported for the first time in Brazil. Additionally, the absence of the former species infecting hosts where the widespread and generalist Sy. (D.) nasuta was found may indicate that Sy. (D.) resticula have either been overlooked in previous studies or that this nematode is rare indeed. Finally, the geographical distribution of Su. halli is greatly expanded and now includes the Americas. Aspects related to the dispersion of these parasites, both in migrating and native hosts, are briefly discussed.

On the role of dauer in the adaptation of nematodes to a parasitic lifestyle.

Nematodes are presumably the most abundant Metazoa on Earth, and can even be found in some of the most hostile environments of our planet. Various types of hypobiosis evolved to adapt their life cycles to such harsh environmental conditions. The five most distal major clades of the phylum Nematoda (Clades 8-12), formerly referred to as the Secernentea, contain many economically relevant parasitic nematodes. In this group, a special type of hypobiosis, dauer, has evolved. The dauer signalling pathway, which culminates in the biosynthesis of dafachronic acid (DA), is intensively studied in the free-living nematode Caenorhabditis elegans, and it has been hypothesized that the dauer stage may have been a prerequisite for the evolution of a wide range of parasitic lifestyles among other nematode species. Biosynthesis of DA is not specific for hypobiosis, but if it results in exit of the hypobiotic state, it is one of the main criteria to define certain behaviour as dauer. Within Clades 9 and 10, the involvement of DA has been validated experimentally, and dauer is therefore generally accepted to occur in those clades. However, for other clades, such as Clade 12, this has hardly been explored. In this review, we provide clarity on the nomenclature associated with hypobiosis and dauer across different nematological subfields. We discuss evidence for dauer-like stages in Clades 8 to 12 and support this with a meta-analysis of available genomic data. Furthermore, we discuss indications for a simplified dauer signalling pathway in parasitic nematodes. Finally, we zoom in on the host cues that induce exit from the hypobiotic stage and introduce two hypotheses on how these signals might feed into the dauer signalling pathway for plant-parasitic nematodes. With this work, we contribute to the deeper understanding of the molecular mechanisms underlying hypobiosis in parasitic nematodes. Based on this, novel strategies for the control of parasitic nematodes can be developed.

Practical guide for microscopic identification of infectious gastrointestinal nematode larvae in sheep from Sardinia, Italy, backed by molecular analysis.

BACKGROUND: Gastrointestinal nematodes (GIN) are ubiquitous in small ruminant farming, representing a major health and production concern. Given their differences in pathogenicity and the current problems regarding anthelmintic resistance, specific diagnosis of GIN is of significant importance. At present, the most widely applied method for this entails culture and microscopic analysis of third-stage larvae, allowing for identification at least to the genus level. Overall, a variety of keys for microscopic analysis have been published, showing substantial variation. Given this fact, this study aimed to produce a practical and updated guide for the identification of infective ovine GIN larvae. METHODS: Using existing keys and protocols, a total of 173larvae of the most common species/genera of ovine GIN from pooled faecal samples from Sardinia (Italy) were identified and extracted, and further individual molecular identification was performed. Morphometric and morphological data as well as high-quality photographs were collected and combined to produce the final guide. RESULTS: GIN microscopically and molecularly identified during this research include Trichostrongylus spp., Teladorsagia circumcincta, Haemonchus contortus, Cooperia curticei, and Chabertia ovina. Based on microscopic analysis, 73.5% of the larvae were correctly identified. Based on sheathed tail length, 91.8% were correctly classified into their respective preliminary groups. CONCLUSIONS: It is crucial for the microscopic identification of infectious GIN larvae to examine each larva in its entirety and thus to take multiple characteristics into account to obtain an accurate diagnosis. However, a preliminary classification based on sheathed tail length (resulting in three groups: A, short; B, medium; C, long) was found to be effective. Further identification within group A can be achieved based on the presence of a cranial inflexion, caudal tubercles and full body measurements (Trichostrongylus spp. < 720 µm, T. circumcincta ≥ 720 µm). Larvae within group B can be differentiated based on sheathed tail morphometry (H. contortus > 65 µm, C. curticei ≤ 65 µm), the presence of cranial refractile bodies, total body length measurements (H. contortus ≤ 790 µm, C. curticei > 790 µm) and shape of the cranial extremity. Finally, all characteristics proposed for the differentiation between Oesophagostomum spp. and C. ovina larvae (group C) were found to have considerable restrictions.

The identification and semi-quantitative assessment of gastrointestinal nematodes in faecal samples using multiplex real-time PCR assays.

BACKGROUND: The diagnosis of gastrointestinal nematode (GIN) infections in ruminants is routinely based on morphological/morphometric analysis of parasite specimens recovered by coprological methods, followed by larval culture (LC) techniques. Such an approach is laborious, time-consuming, requires a skilled expert, and moreover suffers from certain limitations. Molecular tools are able to overcome the majority of these issues, providing accurate identification of nematode species and, therefore, may be valuable in sustainable parasite control strategies. METHODS: Two multiplex real-time polymerase chain reaction (PCR) assays for specific detection of five main and one invasive GIN species, including an internal amplification control to avoid false-negative results, were designed targeting SSU rRNA and COI genetic markers, as well as established ITS1/2 sequences. The assays were optimized for analysis of DNA extracted directly from sheep faeces and verified for Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus colubriformis, Nematodirus battus, Chabertia ovina, and Ashworthius sidemi. Semi-quantitative evaluation of infection intensity was enabled using a plasmid construct and a dilution series of sheep faeces with a known number of nematode eggs. Assays were tested on 44 individually collected faecal samples from three farms, and results were compared to those from faecal egg counts (FEC) using the concentration McMaster technique and LC. RESULTS: Multiplex real-time PCR assays showed great specificity to target nematodes. During the analysis of faecal samples, the assays proved to have higher sensitivity in strongylid-type egg detection over FEC by revealing three false-negative samples, while showing moderate agreement in evaluation of infection intensity. The multiplex assays further clarified GIN species identification compared to LC, which had confused determination of Teladorsagia spp. for Trichostrongylus spp. CONCLUSIONS: Our multiplex assays proved to be a rapid and accurate approach enabling simultaneous and reliable GIN species identification from faeces and semi-quantitative estimation of the number of eggs present. This approach increases diagnostic value and may add a high degree of precision to evaluation of anthelmintic efficacy, where it is important to identify species surviving after treatment.

Non-lethal effects of entomopathogenic nematode infection.

Entomopathogenic nematodes are typically considered lethal parasites of insect hosts. Indeed they are employed as such for biological control of insect pests. The effects of exposure to entomopathogenic nematodes are not strictly limited to mortality, however. Here we explore non-lethal effects of exposure to entomopathogenic nematodes by introducing the relatively non-susceptible pupal stage of Delia antiqua to thirteen different strains. We specifically chose to inoculate the pupal stage because it tends to be more resistant to infection, yet resides in the soil where it could come into contact with EPN biological control agents. We find that there is no significant mortality at the pupal stage, but that there are a host of strain-dependent non-lethal effects during and after the transition to adulthood including altered developmental times and changes in risk of death compared to controls. We also find that exposure to specific strains can reduce risk of mortality. These results emphasize the strain-dependent nature of entomopathogenic nematode infection and highlight the positive and negative ramifications for non-lethal effects for biological control of insect pests. Our work emphasizes the need for strain-specific screening of biological control agents before wide-spread adoption.