Nematode eggshells: A new anatomical and terminological framework, with a critical review of relevant literature and suggested guidelines for the interpretation and reporting of eggshell imagery.

A literature review for a recent ultrastructural study of a trichinelloid eggshell revealed consistently occurring errors in the literature on nematode eggshell anatomy. Examples included nematodes of medical, veterinary, and agricultural importance in several orders. Previous researchers had warned of some of these errors decades ago, but a comprehensive solution was not offered until 2012 when a clarifying new anatomical and developmental interpretation of nematode eggshells was proposed by members of the Caenorhabditis elegans Research Community. However, their findings were explained using arcane acronyms and technical jargon intended for an audience of experimental molecular geneticists, and so their papers have rarely been cited outside the C. elegans community. Herein we (1) provide a critical review of nematode eggshell literature in which we correct errors and relabel imagery in important historical reports; (2) describe common reporting errors and their causes using language familiar to researchers having a basic understanding of microscopy and nematode eggs; (3) recommend a new hexalaminar anatomical and terminological framework for nematode eggshells based on the 2012 C. elegans framework; and (4) recommend new unambiguous terms appropriate for the embryonated/larvated eggs regularly encountered by practicing nematodologists to replace ambiguous or ontogenetically restricted terms in the 2012 C. elegans framework. We also (5) propose a resolution to conflicting claims made by the C. elegans team versus classical literature regarding Layer #3, (6) extend the C. elegans hexalaminar framework to include the polar plugs of trichinelloids, and (7) report new findings regarding trichinelloid eggshell structure.

Title: La coque des œufs des nématodes : un nouveau cadre anatomique et terminologique, avec une revue critique de la littérature pertinente et des lignes directrices suggérées pour l'interprétation et la communication de l'imagerie des coques des œufs. Abstract: Une revue de la littérature pour une étude ultrastructurale récente de la coque de l'œuf d'un trichinelloïde a révélé des erreurs récurrentes dans la littérature sur l'anatomie de la coque de l'œuf des nématodes. Les exemples comprenaient des nématodes d'importance médicale, vétérinaire et agricole dans plusieurs ordres. Des chercheurs avaient mis en garde contre certaines de ces erreurs il y a des décennies, mais une solution complète n'a été proposée qu'en 2012, lorsqu'une nouvelle interprétation anatomique et développementale clarifiant la structure des coques des œufs de nématodes a été proposée par des membres de la communauté de recherche de Caenorhabditis elegans. Cependant, leurs découvertes ont été expliquées à l'aide d'acronymes mystérieux et d'un jargon technique destiné à un public de généticiens moléculaires expérimentaux, et leurs articles ont donc rarement été cités en dehors de la communauté de C. elegans. Ici, nous (1) fournissons une revue critique de la littérature sur les coques des œufs de nématodes dans laquelle nous corrigeons les erreurs et réétiquetons les images dans des rapports historiques importants; (2) décrivons les erreurs de description courantes et leurs causes en utilisant un langage familier aux chercheurs ayant une compréhension de base de la microscopie et des œufs de nématodes; (3) recommandons un nouveau cadre anatomique et terminologique hexalaminaire pour les coques des œufs de nématodes basé sur le cadre de C. elegans de 2012; et (4) recommandons de nouveaux termes non ambigus appropriés pour les œufs embryonnés/larvés régulièrement rencontrés par les spécialistes de nématodes en exercice pour remplacer les termes ambigus ou à restriction ontogénétique dans le cadre de C. elegans de 2012. Nous proposons également (5) une résolution des affirmations contradictoires de l'équipe C. elegans par rapport à la littérature classique concernant la couche 3, (6) étendons le cadre hexalaminaire de C. elegans pour inclure les bouchons polaires des trichinelloïdes, et (7) signalons de nouvelles découvertes concernant la structure de la coque des œufs des trichinelloïdes.

Pharyngodon mamillatus (Nematoda: Pharyngodonidae) in the ocellated skink Chalcides ocellatus (Squamata: Scincidae) from Egypt: Light and scanning electron microscopic studies.

Twenty-one Pharyngodon mamillatus were recovered from the large intestine of 4 out of 12 ocellated skink Chalcides ocellatus collected from the South Sinai desert, Egypt. The prevalence of infection was 33.33% (intensity 2-4). The recovered pharyngodonid was re-described based on light and scanning electron microscopy. According to the morphology, it was characterized by regular transverse annulations extending from the posterior margin of the lips to the level of the anus, triangular mouth opening was surrounded by three large bilobed lips possessing three papillae, oval-shaped excretory pore, slit-like vulva opening, and slit-like anal ventral opening at the posterior end which terminates with a tail filament. Here, the morphometric measurements of adult female P. mamillatus were given in this study. It has no intraspecific morphological variations from the previously described P. mamillatus, but it appeared with small range on biometric features. These variations may be accounted by host/environment attributes since they exhibit wide host and geographical spectra. The present study was applied for detailed morphological knowledge of the recovered P. mamillatus for strengthening their taxonomic validity. HIGHLIGHTS: The Pharyngodon mamillatus naturally parasite of the intestinal region of Chalcides ocellatus. The present study clarifies the morphological key for the description of P. mamillatus.

Unravelling the anthelmintic bioactives from Jasminum grandiflorum L. subsp. Floribundum adopting in vitro biological assessment.

ETHNOPHARMACOLOGICAL RELEVANCE: Jasminum grandiflorum L. is a medicinal plant widely used in the traditional system of Medicine as an anthelmintic in ringworm infections, for treating ulcers, stomatitis, skin diseases, and wounds. AIM OF THE STUDY: The emergence of resistance by different parasites to currently used chemicals has been reported. There are increasing needs for more effective and safer parasiticides. Therefore, the current study was designed to investigate the methanolic extract of the aerial parts of J. grandiflorum subsp. Floribundum (JGTE) to confirm its traditional uses as anthelmintic through a bioassay-guided fractionation and isolation of the active components with anthelmintic activity. MATERIALS AND METHODS: The JGTE was partitioned into dichloromethane (DCM-F) and n-butanol (BuOH-F) fractions. The JGTE, fractions, and the isolated compounds were tested in vitro for their anthelmintic activity using two nematodes; one larval stage of cestode and one arthropod. Four major compounds were isolated from the most active fraction (BuOH-F) including two flavonoids and two secoirridoid glycosides, identified as kaempferol-3-O-neohesperoside (1), rutin (2), oleuropein (3), and ligstroside (4). RESULTS: Among the isolated compounds from most active fraction (BuOH-F), rutin (2) displayed the highest anthelmintic activity in a dose-dependent activity with IC50 of 41.04 µg/mL against H. muscae adult worm, followed by ligstroside (4) with IC50 of 50.56 µg/mL. CONCLUSIONS: These findings could advocate the traditional use of J. grandiflorum L. and provide further insight into the anthelmintic activity of flavonoids.

New data on eggshell structure of capillariid species: a SEM perspective.

Capillariidae is a group of nematode parasites of vertebrates with a complex taxonomy. The structure of the eggshell, which was indicated as the most important characteristic for identification of genus or species through eggs, is very diverse among genera. The visualization and characterization of eggshell by light microscopy (LM) are a challenging task since different planes of the egg surface are needed. Nevertheless, categories of eggshell ornamentation were proposed by LM: smooth, punctuated, reticulated type I, and reticulated type II. The present study aimed to characterize the eggshell structure of Capillariidae species, parasites of mammals and avians, deposited in a helminthological collection using scanning electron microscopy (SEM). Institutional Biological Collections are taxonomic repositories of specimens described and strictly identified at the species level by systematics specialists. SEM eggshell images were obtained from 12 species belonging to 5 genera (Aonchotheca, Baruscapillaria, Capillaria, Echinocoleus, Eucoleus) and compared to their respective LM images. Eggshell patterns observed using SEM were associated categories of eggshell ornamentation previously proposed by LM images. The SEM data indicate that eggshell categories are not in agreement with capillariid genera or sites of infection. However, the study provides previously unknown SEM eggshell information from curated species, which contributes with a specific and supplementary taxonomic feature at the species level of Capillariidae.

Some nematodes from eels (Anguilliformes: Anguillidae) in Japan, with descriptions of two new species.

Recent occasional examinations of two species of eels (Anguilliformes: Anguillidae) in Japan, the Japanese eel Anguilla japonica Temminck & Schlegel from central Japan and the giant mottled eel Anguilla marmorata Quoy & Gaimard from southern Japan, respectively, revealed the following four species of gastrointestinal parasitic nematodes: Cucullanus filiformis Yamaguti, 1935 (Cucullanidae) from A. japonica; Paraquimperia japonica n. sp. (Quimperiidae) from A. japonica (type-host) and A. marmorata; Heliconema anguillae Yamaguti, 1935 (Physalopteridae) from A. marmorata (new host record); and Spinitectus anguillae n. sp. (Cystidicolidae) from A. japonica (type-host) and A. marmorata. Specimens of all species are described based on light and scanning electron microscopical examinations. Paraquimperia japonica n. sp. is mainly characterised by the presence of a ventral sucker, spicules 210-231 µm long and by narrow cervical alae, whereas S. anguillae n. sp. by the number (29-36) of cuticular spines in the first ring, the length of the left spicule (351 µm) and the structure (without polar caps, filaments or lateral swellings), and the size of eggs (36-42 × 21-27 µm).

Structural insights into the fungi-nematodes interaction mediated by fucose-specific lectin AofleA from Arthrobotrys oligospora.

Fungal lectin can bind specific carbohydrate structures of the host and work in recognition and adhesion or as a toxic factor. AofleA, as a fucose-specific lectin from widely studied nematode predatory fungus Arthrobotrys oligospora, possibly plays a key role in the event of capturing nematodes, but the mechanism remains unknown. Here we report the crystal structure of AofleA, which exists as a homodimer with each subunit folds as a six-bladed ß-propeller. Our structural and biological results revealed that three of the six putative binding sites of AofleA had fucose-binding abilities. In addition, we found that AofleA could bind to the pharynx and intestine of the nematode in a fucose-binding-dependent manner. Our results facilitate the understanding of the mechanism that fucose-specific lectin mediates fungi-nematodes interaction, and provide structural information for the development of potential applications of AofleA.

New records of cucullanid nematodes from marine fishes off New Caledonia, with descriptions of five new species of Cucullanus (Nematoda, Cucullanidae).

Recent examinations of cucullanid nematodes (Cucullanidae) from marine fishes off New Caledonia, collected in the years 2004-2009, revealed the presence of the following five new species of Cucullanus Müller, 1777, all parasitic in Perciformes: Cucullanus variolae n. sp. from Variola louti (type host) and V. albimarginata (both Serranidae); Cucullanus acutospiculatus n. sp. from Caesio cuning (Caesionidae); Cucullanus diagrammae n. sp. from Diagramma pictum (Haemulidae); Cucullanus parapercidis n. sp. from Parapercis xanthozona (type host) and P. hexophtalma (both Pinguipedidae); and Cucullanus petterae n. sp. from Epinephelus merra (type host) and E. fasciatus (both Serranidae). An additional congeneric species, Cucullanus bioccai Orecchia et Paggi, 1987 was recorded from Mugil cephalus (Mugilidae, Mugiliformes) (first record in the Pacific Ocean) and Cucullanus sp. (only female) was found in Arothron manilensis (Tetraodontidae, Tetraodontiformes). Furthermore, two known cucullanid species, Dichelyne (Cucullanellus) branchiostegi (Yamaguti, 1941) in Branchiostegus wardi (Malacanthidae, Perciformes) (new host and geographical records) and Dichelyne (Cucullanellus) bodiani Moravec et Justine, 2019 in Bodianus busellatus (new host) and B. perditio (both Labridae, Perciformes), were found; Dichelyne (Cucullanellus) sp. (only females) coinfecting the latter host may represent an unknown species. Most species are described based on light and electron microscopical studies. The specimens described by Xu et al., 2017 as Cucullanus bourdini Petter et Le Bel, 1992 from Caesio xanthonota (Caesionidae) in the Taiwan Strait are considered to represent a new species, for which the name Cucullanus sinensis n. sp. is proposed.

TITLE: Nouvelles mentions de nématodes Cucullanidae de poissons marins de Nouvelle-Calédonie, avec description de cinq nouvelles espèces de Cucullanus (Nematoda, Cucullanidae). ABSTRACT: L'étude récente de nématodes Cucullanidae de poissons marins du large de la Nouvelle-Calédonie, collectés dans les années 2004-2009, a révélé la présence des cinq nouvelles espèces suivantes de Cucullanus Müller, 1777, toutes parasites de Perciformes : Cucullanus variolae n. sp. de Variola louti (hôte type) et V. albimarginata (tous deux Serranidae) ; Cucullanus acutospiculatus n. sp. de Caesio cuning (Caesionidae) ; Cucullanus diagrammae n. sp. de Diagramma pictum (Haemulidae) ; Cucullanus parapercidis n. sp. de Parapercis xanthozona (hôte type) et P. hexophtalma (tous deux Pinguipedidae) ; et Cucullanus petterae n. sp. d'Epinephelus merra (type hôte) et E. fasciatus (tous deux Serranidae). Une autre espèce congénérique, Cucullanus bioccai Orecchia et Paggi, 1987 est signalée chez Mugil cephalus (Mugilidae, Mugiliformes) (première mention dans l'océan Pacifique) et Cucullanus sp. (seulement femelle) a été trouvé chez Arothron manilensis (Tetraodontidae, Tetraodontiformes). De plus, deux espèces de Cucullanidae connues, Dichelyne (Cucullanellus) branchiostegi (Yamaguti, 1941) chez Branchiostegus wardi (Malacanthidae, Perciformes) (nouvel hôte et nouvelle mention géographique) et Dichelyne (Cucullanellus) bodiani Moravec et Justine, 2019 chez Bodianus busellatus (nouvel hôte) et B. perditio (tous deux Labridae, Perciformes), ont été trouvées ; Dichelyne (Cucullanellus) sp. (seulement femelles) co-infectant ce dernier hôte peut représenter une espèce inconnue. La plupart des espèces sont décrites sur la base d'études au microscope optique et électronique. Les spécimens décrits par Xu et al., 2017 comme Cucullanus bourdini Petter et Le Bel, 1992 chez Caesio xanthonota (Caesionidae) dans le détroit de Taiwan sont considérés comme représentant une nouvelle espèce, pour laquelle le nom Cucullanus sinensis n. sp. est proposé.

First record of Pseudoterranova decipiens (Nematoda, Anisakidae) infecting the Red spot emperor Lethrinus lentjan in the Red Sea / Primeiro registro de Pseudoterranova decipiens (Nematoda, Anisakidae) infectando o imperador da mancha vermelha Lethrinus lentjan no Mar Vermelho

Abstract The current parasitological study was carried out to investigate helminth parasites infecting the Red spot emperor Lethrinus lentjan inhabiting Hurghada City at the Gulf of Suez, Red Sea, Egypt. Third-stage larvae of nematode parasite was isolated from the intestine as well as body cavity of the examined fish. Light and scanning electron microscopy revealed that this parasite belonged to Anisakidae family within the genus Pseudoterranova. The present species is named Pseudoterranova decipiens based on the presence of triangular mouth aperture with prominent boring teeth and soft swellings of the cuticle, long muscular esophagus, ventrally excretory pore, and narrow transverse slit of anal opening followed by a short mucron. The morphological characteristics of this species were confirmed by molecular analysis of 18S rDNA gene region of the present parasite. It demonstrated a close identity ≥89% with taxa under family Anisakidae, 85% with Raphidascarididae, and 79-84% with Toxocaridae. A preliminary genetic comparison between gene sequence of the present parasite and other oxyurid species placeed it as a putative sister taxon to other Pseudoterranova decipiens described previously. This study demonstrated that the 18S rDNA gene region of Pseudoterranova decipiens yielded a unique sequence that confirmed its taxonomic position in Anisakidae.

Resumo O presente estudo parasitológico foi realizado para investigar os helmintos parasitos que infectam o peixe imperador Lethrinus lentjan, que habita a cidade de Hurghada no Golfo de Suez, Mar Vermelho, no Egito. Larvas de terceiro estágio de parasitos nematoides foram isoladas do intestino e da cavidade do corpo do peixe examinado. Microscopia eletrônica de luz e de varredura revelou que este parasita pertence à família Anisakidae dentro do gênero Pseudoterranova. A espécie atual é denominada Pseudoterranova decipiens baseada na presença de abertura triangular da boca com dentes proeminentes chatos e inchaços moles da cutícula, esôfago muscular longo, poro ventralmente excretor e fenda transversal estreita da abertura anal seguida por um mucron curto. As características morfológicas desta espécie foram confirmadas pela análise molecular da região do gene 18S rDNA do presente parasito. Demonstrou uma identidade próxima ≥89% com taxa sob família Anisakidae, 85% com Raphidascarididae, e 79-84% com Toxocaridae. Uma comparação genética preliminar entre a sequência genética do presente parasito e outras espécies de oxiurídeos coloca-o como um taxon irmão putativo para outros Pseudoterranova descritos anteriormente. Este estudo demonstra que a região do gene 18S rDNA de Pseudoterranova decipiens produz uma sequência única que confirma sua posição taxonômica em Anisakidae.

First record of Pseudoterranova decipiens (Nematoda, Anisakidae) infecting the Red spot emperor Lethrinus lentjan in the Red Sea.

The current parasitological study was carried out to investigate helminth parasites infecting the Red spot emperor Lethrinus lentjan inhabiting Hurghada City at the Gulf of Suez, Red Sea, Egypt. Third-stage larvae of nematode parasite was isolated from the intestine as well as body cavity of the examined fish. Light and scanning electron microscopy revealed that this parasite belonged to Anisakidae family within the genus Pseudoterranova. The present species is named Pseudoterranova decipiens based on the presence of triangular mouth aperture with prominent boring teeth and soft swellings of the cuticle, long muscular esophagus, ventrally excretory pore, and narrow transverse slit of anal opening followed by a short mucron. The morphological characteristics of this species were confirmed by molecular analysis of 18S rDNA gene region of the present parasite. It demonstrated a close identity ≥89% with taxa under family Anisakidae, 85% with Raphidascarididae, and 79-84% with Toxocaridae. A preliminary genetic comparison between gene sequence of the present parasite and other oxyurid species placeed it as a putative sister taxon to other Pseudoterranova decipiens described previously. This study demonstrated that the 18S rDNA gene region of Pseudoterranova decipiens yielded a unique sequence that confirmed its taxonomic position in Anisakidae.

Nematodes found in Nile crocodiles in the Kruger National Park, South Africa, with redescriptions of Multicaecum agile (Wedl, 1861) (Heterocheilidae) and Camallanus kaapstaadi Southwell & Kirshner, 1937 (Camallanidae).

Sixteen Nile crocodiles were collected in the Kruger National Park, South Africa and vicinity during 2010 and 2011. A total of 11 nematode species representing six families were recovered. Heterocheilids were the dominant group, comprising five species, with Dujardinascaris madagascariensis (Chabaud & Caballero, 1966) being the most prevalent (75%), followed by Ingwenascaris sprenti Junker & Mutafchiev, 2017 (68.8%), which was also the second most numerous nematode. While less prevalent (31.3%), Typhlophoros kwenae Junker & Mutafchiev, 2017 was the most abundant species. Micropleura huchzermeyeri Junker & Mutafchiev, 2017 (Micropleuridae) was collected from five crocodiles and Crocodylocapillaria sp. (Capillariidae) occurred in a single host. Three nematodes, Camallanus kaapstaadi Southwell & Kirshner, 1937, Spirocamallanus sp. (both Camallanidae) and Ascarophis sp. (Cystidicolidae), are considered accidental infections, likely ingested with the hosts' prey. Our findings of D. dujardini (Travassos, 1920), D. madagascariensis and Multicaecum agile (Wedl, 1861) in South Africa constitute new geographical records. Crocodylocapillaria sp. represents a new host and geographical record, while T. kwenae, I. sprenti and M. huchzermeyeri have been described as new species during the course of this survey. Multicaecum agile is here redescribed based on light and scanning electron microscopy. Previously undescribed morphological characters of C. kaapstaadi, typically a parasite of Xenopus spp. (Amphibia: Pipidae), but here found in two Nile crocodiles, are also presented.

First report of filarial nematodes in free-living pitheciid primates.

Although little studied, infections with nematodes of the Onchocercidae Leiper, 1911, predominated by the genera Dipetalonema Diesing, 1861 and Mansonella Faust, 1929, are frequent in wild primates and human populations in the Neotropical forest areas. This study reports natural infections with Dipetalonema freitasi Bain, Diagne & Muller, 1987 and D. gracile (Rudolphi, 1809) in two free-living species of pitheciid primates, extending the known geographical distribution of these species to the forest of the Peruvian Amazon. Adult worms were recovered from the thoracic and abdominal cavities of two species of monkeys, Pithecia monachus monachus (É. Geoffroy Saint-Hilaire) and Cacajao calvus ucayalii (Thomas) (Primates: Pitheciidae), collected along the Yavari-Mirin River basin and analysed via light and scanning electron microscopy. Both host species represent new host records for D. freitasi and D. gracile. Morphometric data are also presented for the sampled filarial worms in addition to morphological details obtained through light and electron microscopy examination of D. freitasi specimens.

Morphological and genetic characterization of Syphabulea tjanschani () (Nematoda: Oxyuridae), with phylogenetic position of Syphabulea in Oxyuridae.

Pinworms are important parasitic nematodes in animals and humans, and many species are of medical and veterinary importance. The genus Syphabulea is a poorly known group of pinworms. The systematic position of Syphabulea in Oxyuridae remains unclear. Moreover, there is still a paucity of detailed information on some morphological aspects of the type species S. tjanschani (Ablasov, 1962). In the present study, the detailed morphology of S. tjanschani was studied using light microscopy and, for the first time, scanning electron microscopy. The results revealed three traits useful for the characterization of the species including: cuticular depression around excretory pore, non-prominent labial teeth and number of adhesive ridges in three ventral mammelons. The ribosomal [small ribosomal DNA (18S), internal transcribed spacer (ITS) and large ribosomal DNA (28S)] and mitochondrial [cytochrome c oxidase subunit 1 (cox1)] target regions of S. tjanschani were also firstly amplified and sequenced for future use in the molecular identification of this poorly known species. In addition, in order to investigate the systematic position of Syphabulea in Oxyuridae, the phylogenetic analyses of representatives of the Oxyuridae were performed using maximum likelihood (ML) and maximum parsimony (MP) methods based on 18S, 28S and 18S + 28S + cox1 sequence data, respectively. The phylogenetic results based on different sequence data all supported the genus Syphabulea to be a member of the subfamily Syphaciinae. The phylogenetic analysis based on 28S sequence data also showed Syphabulea as sister to the genus Syphatineria. Our present study represents the first attempt to resolve the systematic position of the genus Syphabulea using phylogenetic analyses based on molecular data.

Diaphanocephalus galeatus (Nematoda: Diaphanocephalidae), a poorly known parasite of lizards: redescription, first genetic characterization and a revision of its congeners from Brazil.

Diaphanocephalus galeatus collected from the small intestine of the lizard Dracaena paraguayensis in the Pantanal wetlands, State of Mato Grosso do Sul, Brazil, is redescribed. Genetic characterization and observations using scanning electron microscopy (SEM) were performed for the first time. The vouchers of D. galeatus and the type specimens of its congeners, deposited in the Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC), were consulted. Light and SEM observations revealed several undescribed features of D. galeatus, i.e. structure of the cephalic end and of the buccal capsule, position and morphology of deirids, presence of phasmids in females and presence of unpaired papilla on the membranous projection that covers the genital cone in males. After observation of the specimens deposited in the helminthological collection, D. jacuruxi is considered a synonym of D. galeatus, and D. diesingi, despite its incomplete description, is tentatively retained as valid due to the poor condition of the type material. The results also indicated low host specificity of D. galeatus, contradicting previous assertions. Genetic comparisons using patristic distances and phylogenetic trees generated from sequences of the 28S rRNA nuclear gene indicated that D. galeatus is closer to the taxa within Ancylostomatoidea and Strongyloidea than any lineage of Metastrongyloidea or Trichostrongyloidea. However, most of the nodal supports were low. Based on the genetic and morphological characterization, the validity of D. galeatus was confirmed. These data may serve for further comparative approaches for different populations of the parasite, from different hosts in different geographical areas, mitigating taxonomic confusions.

Nematode capilaridae in the tongue of Cerdocyon thous of free life in Brazil / Nematoide capilaridae na língua de Cerdocyon thous de vida livre no Brasil

Abstract Studies of helminths in road-killed wild animals are still uncommon but may provide promising results since they can identify the parasites in juvenile and adult hosts and meet the recommendations of current discussions on bioethics that prioritize alternative methods for the use of animals. This study evaluated three Cerdocyon thous individuals that were donated after dying from being run over. Two of them had small nematode adults in the epithelial and connective tissues of the tongue. The diagnosis was based on the presence of eggs, observed in histological sections, and morphological characteristics of the nematodes in the tongue. Morphologically, this nematode has a body with transverse grooves, simple mouth opening and no lips, esophagus and stichosome with stichocytes and bacillary bands along the body, which is characteristic morphology of the family Capilariidae and genus Capillaria . The presence of this nematode in the tongue of C. thous is an extremely important fact that contributes to what is known about the biodiversity of zoonotic parasites in wild canid populations. However, an explanation for these findings remains unclear because, until now, this has not been observed in the biological cycle of the species.

Resumo O estudo de helmintos de animais selvagens vitimados por atropelamento, ainda é uma prática pouco comum, porém pode ser promissor, uma vez que permite a identificação de helmintos a partir de hospedeiros juvenis e adultos, e atendem às atuais discussões em bioética, as quais priorizam métodos alternativos para o uso de animais. Foram avaliados 3 exemplares de Cerdocyon thous provenientes de doação pós-óbito por atropelamento. Dois deles apresentaram pequenos nematoides adultos inseridos nos tecidos epitelial e conjuntivo da língua. O diagnóstico foi baseado na análise de cortes histológicos que evidenciaram a presença de ovos no interior do nematoide e nas características morfológicas gerais do mesmo. Morfologicamente estes nematoides apresentavam corpo com estrias transversais, abertura bucal simples e lábios ausentes, esôfago trichuroiude com a presença de esticócitos e bandas bacilares ao longo do corpo, característica morfologicamente sugestiva da família Capillariidae e do gênero Capillaria . A presença de nematoides inseridos na língua de C. thous é um fato de extrema importância para o conhecimento da biodiversidade de parasitos zoonóticos circulantes presentes em populações selvagens de canídeos. Porém, permanece obscura a explicação para tais achados, uma vez que tal evento não foi observado no ciclo biológico das espécies até o momento.

Cucullanus marajoara n. sp. (Nematoda: Cucullanidae), a parasite of Colomesus psittacus (Osteichthyes: Tetraodontiformes) in the Marajó, Brazil / Cucullanus marajoara n. sp. (Nematoda: Cucullanidae), um parasito de Colomesus psittacus (Osteichthyes: Tetraodontiformes) no Marajó, Brasil

Abstract Cucullanus marajoara n. sp. (Cucullanidae) is reported to parasitize Colomesus psittacus (Tetraodontiformes), which is a fish species from the Marajó Archipelago, state of Pará, estuarine region of the Brazilian Amazon. The new species differs from similar species by the presence of a protruding upper lip on the cloacal opening, the distribution of the cloacal papillae: five pre-cloacal papillae pairs and 5 are ventral and located posteriorly to the pre-cloacal sucker and an unpaired papilla is located on the upper cloacal lip and five post-cloacal pairs, and a pair of lateral phasmids located between papillae pairs. Additionally, Cucullanus marajoara n. sp. is compared to other species of this genus described in Brazil, particularly Cucullanus ageneiosus and Cucullanus oswaldocruzi, which both occur in the same zoogeographic region of this study but parasitize fish of a different order (Siluriformes). Cucullanus dodsworthi and Cucullanus brevicaudatus are the only described species parasitizing fish of the order Tetradontiformes in Brazil, and the new species differs from these species by the distribution of the cloacal papillae and the host habitat. The description of Cucullanus marajoara n. sp. adds data to the biodiversity of described parasites that parasitize Tetradontiformes of the estuarine ichthyofauna in the Brazilian Amazon.

Resumo Cucullanus marajoara n. sp. (Cucullanidae) é descrita parasitando Colomesus psittacus (Tetraodontiformes) peixe do arquipélago de Marajó, Estado do Pará, região estuarina da Amazônia brasileira. A nova espécie difere de seus congêneres por apresentar lábio superior da abertura cloacal protrudente e na distribuição de papilas cloacais: 5 pares pré-cloacais, e uma papila não pareada no lábio superior da cloaca e 5 pares pós-cloacais, um par de fasmídeos laterais. Além disso, Cucullanus marajoara n. sp. é comparada com outras espécies do gênero descritas no Brasil, em especial Cucullanus ageneiosus e Cucullanus oswaldocruzi, ambos parasitos de peixes da ordem Siluriformes; porém, ocorrendo na mesma região zoogeográfica deste estudo, diferem da ordem do hospedeiro da nova espécie (Tetraodontiformes). Cucullanus dodsworthi e Cucullanus brevicaudatus são as únicas espécies descritas parasitando peixe da ordem Tetradontiformes no Brasil, e a nova espécie difere dessas na distribuição das papilas cloacais e habitat dos hospedeiros. Cucullanus marajoara n. sp. adiciona dados a biodiversidade de parasitos descritos parasitando Tetradontiformes da ictiofauna estuarina da Amazônia brasileira.

Cucullanus marajoara n. sp. (Nematoda: Cucullanidae), a parasite of Colomesus psittacus (Osteichthyes: Tetraodontiformes) in the Marajó, Brazil.

Cucullanus marajoara n. sp. (Cucullanidae) is reported to parasitize Colomesus psittacus (Tetraodontiformes), which is a fish species from the Marajó Archipelago, state of Pará, estuarine region of the Brazilian Amazon. The new species differs from similar species by the presence of a protruding upper lip on the cloacal opening, the distribution of the cloacal papillae: five pre-cloacal papillae pairs and 5 are ventral and located posteriorly to the pre-cloacal sucker and an unpaired papilla is located on the upper cloacal lip and five post-cloacal pairs, and a pair of lateral phasmids located between papillae pairs. Additionally, Cucullanus marajoara n. sp. is compared to other species of this genus described in Brazil, particularly Cucullanus ageneiosus and Cucullanus oswaldocruzi, which both occur in the same zoogeographic region of this study but parasitize fish of a different order (Siluriformes). Cucullanus dodsworthi and Cucullanus brevicaudatus are the only described species parasitizing fish of the order Tetradontiformes in Brazil, and the new species differs from these species by the distribution of the cloacal papillae and the host habitat. The description of Cucullanus marajoara n. sp. adds data to the biodiversity of described parasites that parasitize Tetradontiformes of the estuarine ichthyofauna in the Brazilian Amazon.

Nematode capilaridae in the tongue of Cerdocyon thous of free life in Brazil.

Studies of helminths in road-killed wild animals are still uncommon but may provide promising results since they can identify the parasites in juvenile and adult hosts and meet the recommendations of current discussions on bioethics that prioritize alternative methods for the use of animals. This study evaluated three Cerdocyon thous individuals that were donated after dying from being run over. Two of them had small nematode adults in the epithelial and connective tissues of the tongue. The diagnosis was based on the presence of eggs, observed in histological sections, and morphological characteristics of the nematodes in the tongue. Morphologically, this nematode has a body with transverse grooves, simple mouth opening and no lips, esophagus and stichosome with stichocytes and bacillary bands along the body, which is characteristic morphology of the family Capilariidae and genus Capillaria . The presence of this nematode in the tongue of C. thous is an extremely important fact that contributes to what is known about the biodiversity of zoonotic parasites in wild canid populations. However, an explanation for these findings remains unclear because, until now, this has not been observed in the biological cycle of the species.

Atomic force microscopy for imaging and nanomechanical characterisation of live nematode epicuticle: A comparative Caenorhabditis elegans and Turbatrix aceti study.

Atomic force microscopy (AFM), a powerful tool in interdisciplinary biomedical research, has been applied here to investigate the surface of live nematodes epicuticle. We have used AFM in PeakForce Tapping non-resonant imaging and nanomechanical characterisation mode to investigate and compare the surface features of epicuticle of two free-living microscopic nematodes, Caenorhabditis elegans and Turbatrix aceti. We have successfully immobilised live anesthetized adult nematodes on glass supports using either layer-by-layer-deposited polyelectrolyte films or bioadhesive coatings, which allowed for imaging the living nematodes in native environment. We have obtained AFM images and corresponding nanomechanical maps of annular rings and furrows, demonstrating the differences in topography and structure between the species. Our results demonstrate that AFM in PeakForce Tapping mode can be used to image and characterise surfaces of relatively-large live immobilised multicellular organisms, which can be further applied to a number of invertebrates.

Rasheedia n. nom. (Nematoda, Physalopteridae) for Bulbocephalus Rasheed, 1966 (a homonym of Bulbocephalus Watson, 1916), with description of Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. from perciform fishes off New Caledonia.

The nematode genus Bulbocephalus Rasheed, 1966 (Nematoda, Physalopteridae) was found to be a homonym of Bulbocephalus Watson, 1916 (Apicomplexa) and, therefore, a new name, Rasheedia n. nom., is proposed to substitute it. Based on light and scanning electron microscope studies of specimens collected from the digestive tract of perciform fishes off New Caledonia, two new species of Rasheedia are described: R. heptacanthi n. sp. from the Cinnabar goatfish Parupeneus heptacanthus (Mullidae) (type host) and Dentex fourmanoiri (Sparidae), and R. novaecaledoniensis n. sp. from the Indian goatfish Parupeneus indicus (Mullidae). These new species are mainly characterized by the number of anterior protrusible oesophageal lobes (two in R. heptacanthi and four in R. novaecaledoniensis), structure of the oesophagus and the lengths of spicules. An amended diagnosis of Rasheedia and a key to species of this genus are provided. Three previously described congeneric species are transferred to Rasheedia as R. deblocki (Le-Van-Hoa, Pham-Ngoc-Khue & Nguyen-Thi-Lien, 1972) n. comb., R. inglisi (Rasheed, 1966) n. comb. and R. pseudupenei (Vassiliadès & Diaw, 1978) n. comb. Cestocephalus Rasheed, 1966 [genus inquirendum], including C. serratus Rasheed, 1966 and C. petterae (Le-Van-Hoa, Pham-Ngoc-Khue & Nguyen-Thi-Lien, 1972) n. comb., should be considered to be separate from Rasheedia. The names Pseudomazzia Bilqees, Ghazi & Haseeb, 2005 and P. macrolabiata Bilqees, Ghazi & Haseeb, 2005, established for a nematode somewhat resembling Rasheedia spp., should be considered nomina dubia. Rasheedia heptacanthi n. sp. and R. novaecaledoniensis n. sp. are the first representatives of the Physalopteridae recorded from fishes in New Caledonian waters.

TITLE: Rasheedia n. nom. (Nematoda, Physalopteridae) pour Bulbocephalus Rasheed, 1966 (homonyme de Bulbocephalus Watson, 1916), avec description de Rasheedia heptacanthi n. sp. et R. novaecaledoniensis n. sp. de poissons perciformes de Nouvelle-Calédonie. ABSTRACT: Le genre de nématode Bulbocephalus Rasheed, 1966 (Nematoda, Physalopteridae) a été trouvé homonyme de Bulbocephalus Watson, 1916 (Apicomplexa) et, par conséquent, un nouveau nom, Rasheedia n. nom., est proposé pour le remplacer. Sur la base d'études en microscopie optique et électronique à balayage de spécimens prélevés dans le tube digestif de poissons perciformes de Nouvelle-Calédonie, deux nouvelles espèces de Rasheedia sont décrites : R. heptacanthi n. sp. chez Parupeneus heptacanthus (Mullidae) (hôte-type) et Dentex fourmanoiri (Sparidae) et R. novaecaledoniensis n. sp. chez Parupeneus indicus (Mullidae). Ces nouvelles espèces sont principalement caractérisées par le nombre de lobes œsophagiens protrusibles antérieurs (deux chez R. heptacanthi et quatre chez R. novaecaledoniensis), la structure de l'œsophage et la longueur des spicules. Un diagnostic modifié de Rasheedia et une clé pour les espèces de ce genre sont fournis. Trois espèces congénériques précédemment décrites sont transférées dans Rasheedia sous les noms R. deblocki (Le-Van-Hoa, Pham-Ngoc-Khué et Nguyen-Thi-Lien, 1972) n. comb., R. inglisi (Rasheed, 1966) n. comb. et R. pseudupenei (Vassiliadès & Diaw, 1978) n. comb. Cestocephalus Rasheed, 1966 [genus inquirendum], incluant C. serratus Rasheed, 1966 et C. petterae (Le-Van-Hoa, Pham-Ngoc-Khué et Nguyen-Thi-Lien, 1972) n. comb., doit être considéré comme distinct de Rasheedia. Les noms Pseudomazzia Bilqees, Ghazi & Haseeb, 2005 et P. macrolabiata Bilqees, Ghazi & Haseeb, 2005, établis pour un nématode ressemblant un peu à Rasheedia spp., devraient être considérés comme nomina dubia. Rasheedia heptacanthi n. sp. et R. novaecaledoniensis n. sp. sont les premiers Physalopteridae rapportés de poissons dans les eaux de Nouvelle-Calédonie.

Motility in the L3 stage is a poor phenotype for detecting and measuring resistance to avermectin/milbemycin drugs in gastrointestinal nematodes of livestock.

Motility is a commonly used in vitro phenotype for assessing anthelmintic activity of candidate compounds, and for detecting anthelmintic resistance in nematodes. Third-stage larvae (L3) of parasitic nematodes are commonly used in motility-based assays because L3 are simple to obtain and can remain viable in storage for extended periods. To improve the measurement of motility of microscopic stages of nematodes, our laboratory developed the Worminator, which quantitatively measures motility of parasites. Using the Worminator, we compared the dose-response characteristics of several avermectin/milbemycin (AM) compounds using L3 from both AM-susceptible and AM-resistant Cooperia spp. (abamectin, doramectin, eprinomectin, ivermectin, moxidectin) and Haemonchus contortus (eprinomectin, ivermectin, moxidectin). Concentrations tested with the Worminator ranged from 0.156 to 40 µM. Differences in EC50 between AM-susceptible and AM-resistant isolates of Cooperia spp. and Haemonchus contortus were small, with resistance ratios ranging from 1.00 to 1.34 for Cooperia spp., 0.99 to 1.65 for Haemonchus contortus. Larval migration inhibition assays were conducted using the same isolates and were equally ineffective for detection of resistance with resistance ratios less than 2.0. These results contrast with those of the Larval Development Assay where we obtained a resistance ratio of 16.48 using the same isolates of Haemonchus contortus. Moreover, even at the highest concentration tested (40 µM), 100% inhibition of motility was never achieved and EC50 for Worminator assays were more than 100× higher than peak plasma levels achieved in vivo following treatment. These data demonstrate that dose-response characteristics for inhibition of motility in L3 of gastrointestinal nematodes of livestock do not significantly differ for AM-susceptible and AM-resistant isolates. These data challenge the suitability of motility as a phenotype for detecting and measuring resistance to AM drugs in gastrointestinal nematodes of livestock.