Scanning electron microscopy of Echinostoma revolutum (Trematoda) during development in the chick embryo and the domestic chick

Fried B. and Fujino T. 1984. Scanning electron microscopy of Echinostoma revolutum (Trematoda) during development in the chick embryo and the domestic chick. International Journal for Parasitology14: 75–81. Scanning electron microscopy (SEM) was used to study the development of chemically excysted metacercariae of Echinostoma revolutum on the chick chorioallantois. SEM studies were also made on preovigerous adults of E. revolutum grown in the domestic chick. During worm development on the chorioallantois the tegument changed from smooth to granular and sensory papillae on the suckers became well-defined. As worms developed on the chorioallantois the cephalic collar spines became thicker and more curved and the tegumentary spines showed marked changes in shape, size and distribution on both ventral and dorsal aspects of the body. Changes in the surface ultrastructure of worms grown on the chorioallantois were essentially similar to those observed in preovigerous worms from chicks.     

Teratological Development in the Cephalic Anatomy of the Nematode Romanomermis culicivorax

Adults of Romanomermis culicivorax obtained from mass cultures were examined by scanning and transmission electron microscopy to determine the organization of their anterior sense organs. The normal pattern apparently consists of two lateral amphids plus six cephalic papillae. Lateral cephalic papillae contain two sense organs, each with a cuticular pore, while subdorsal and subventral papillae have three sense organs, each with a cuticular pore. About 30% of females and 80% of males examined showed aberrant developments in these sense organs. Both cuticle and underlying cells (hypodermis and neurons) are affected; some sense organs are absent while others are incompletely formed. Few aberrant worms were found in a smaller collection of worms reared in lower population densities. Perhaps aberrant forms are examples of teratological development resulting from, or promoted by, conditions used for mass rearing of biological control agents.     

Purification of Onchocerca gibsoni microfilariae

Forsyth K. P., Copeman D. B. and Mitchell G. F. 1982. Purification of Onchocerca gibsoni microfilariae. International Journal for Parasitology12: 53–57. Viable microfilariae from the bovine parasite, Onchocerca gibsoni, were purified to a level suitable for immunochemical analysis and without appreciable loss in numbers by low speed centrifugation on Ficoll-paque. Microfilariae were obtained by incubation of intact O. gibsoni nodules or from worm fragments dissected from nodules. After purification, microfilariae were determined as viable by motility, ability to establish in mice and to incorporate ³⁵S-methionine into large numbers of proteins in vitro as determined by two-dimensional gel electrophoresis and fluorography.     

Isolation of Onchocerca gibsoni tissue from frozen nodules for biochemical use

A new procedure is described which enables gram quantities of adult Onchocerca tissue to be isolated from frozen connective tissue nodules, thus minimizing the risk of enzymatic degradation. Bovine connective tissue nodules containing adult Onchocerca gibsoni worms were obtained from Australia frozen at −70°C and sectioned while still frozen into 3 mm thick slabs. The sections were thawed immediately before use, worm segments removed, rinsed, pelleted, and flash frozen in liquid nitrogen. Quality of the isolated material was demonstrated by the presence of an intact adult epicuticle as determined by electron microscopy, and by the presence of viable uterine larvae and cells. This procedure is applicable to other nodule-forming worms such as Onchocerca volvulus and is suitable for investigations which require the isolation of labile molecules or those present in minute quantities.     

Further studies on Contracaecum spasskii Mozgovoi, 1950 and C. rudolphii Hartwich, 1964 (sensu lato) (Ascaridida: Anisakidae) from piscivorous birds in China

Contracaecum spasskii Mozgovoi, 1950, collected from the great crested grebe Podiceps cristatus (Linnaeus) (Podicipediformes: Podicipedidae), is redescribed using both light and, for the first time, scanning electron microscopy. Contracaecum spasskii differs from its congeners by having marked transverse cuticular annulations, the length of the oesophagus and spicules, the ratio between the intestinal caecum and the ventricular appendix, the number and arrangement of male caudal papillae, and especially by the particular morphology of the lips and interlabia. Some previously unreported morphological features of C. spasskii are also revealed and others corrected. Contracaecum rudolphii Hartwich, 1964 (sensu lato) is also redescribed based on the specimens collected from the great cormorant Phalacrocorax carbo sinensis (Blumenbach) (Pelecaniformes: Phalacrocoracidae) from China. Based on the geographical perspective, the present Chinese material may represent the species C. rudolphii B.     

Scanning Electron Microscope Studies of Steinernema anomali Kozodoi, 1984

This paper presents SEM micrographs of portions of the male, female, and infective-stage juvenile of Steinernema anomali. Included are micrographs of the cephalic and caudal region, spicules, and gubernaculum of the male, the cephalic and vulval region of the female, and the cephalic region of the infective-stage juvenile. Males have six labial and four prominent cephalic papillae and small amphids. There are 11-14 pairs and one single genital papillae; of these, 6-9 pairs are preanal and subventral, one pair preanal, lateral, one pair adanal, and three pairs postanal. Spicules have a short head, a long blade, and a reduced shaft. The distal end is enlarged and bears a dorsal aperture. Gubernaculum much shorter than spicules; cuneus of gubernaculum short and bifurcate anteriorly. Females have six labial and four cephalic papillae and small amphids. Vulva with a thickened posterior lip. Infective juveniles have a smooth head, prominent amphids, and four cephalic papillae. Labial papillae, if present, are not evident.     

New data on the morphology of Philonema oncorhynchi Kuitunen-Ekbaum, 1933 (Nematoda: Dracunculoidea) from the abdominal cavity of Pacific salmon (Oncorhynchus spp.)

The dracunculoid nematode Philonema oncorhynchi Kuitunen-Ekbaum, 1933 is redescribed from specimens collected from three species of anadromous Pacific salmon, i.e. Oncorhynchus keta (Walbaum), O. kisutch (Walbaum) and O. nerka (Walbaum), from Japan, North Pacific Ocean and eastern Russia (Kamchatka), respectively. Scanning electron microscopy revealed the presence of hitherto undescribed cuticular ornamentations on the male caudal end and made it possible to establish the numbers, distribution and shape of cephalic and caudal papillae in the male, which are hardly visible under a light microscope; two horn-like protrusions and a median pre-anal papilla situated on the anterior cloacal lip were not previously described in this species. The necessity of detailed morphological studies on Philonema spp. parasitic in salmonids for the clarification of their taxonomy is stressed.     

Immunity to Onchocerca lienalis microfilariae in mice. II. Effects of sensitization with a range of heterologous species

The model of Onchocerca lienalis microfilariae (mff) injected into CBA and T.O. strains of mice has been used to examine immunity to skin-dwelling microfilariae following exposure to a range of species of helminths. Mice which had received a primary infection with O. cervicalis mff were significantly resistant to challenge with O. lienalis mff (58% reduction relative to challenge controls). Immunization with the uterine contents (eggs and mff) of O. lienalis, O. gutturosa or O. volvulus conferred equivalent levels of protection against challenge with O. lienalis mff (66 to 75%). Similar results were obtained with immunizations in mice that employed either fresh or freeze-killed eggs of O. gutturosa. Significant reductions in the recoveries of O. lienalis mff were also demonstrated following the intraperitoneal implantation of adult male worms of O. gutturosa (30 to 52%), the adults of either sex of Dipetalonema viteae (60%), or after infection with Trichinella spiralis (27 to 81%). Infections with the trematode, Schistosoma mansoni, had a negligible effect on mff recoveries. It is concluded that partial resistance in mice to Onchocerca mff may be stimulated by factors, yet to be determined, that are neither stage nor species-specific.     

Scanning Electron Microscopy of Second-Stage Juvenile Cephalic Morphology in Heterodera glycines Races

Scanning electron microscopy (SEM) was used to compare juvenile cephalic morphology of the five described races of Heterodera glycines. Races 1, 2, 3, and 4 were obtained in the United States and race 5 was obtained from Japan. Differences in the gross morphology o f labial discs; ventral, dorsal and lateral lips; amphidial apertures; and fissures on the labial disc o f some specimens were observed. There was considerable interracial and intraracial variation which precluded separation o f juveniles of H. glycines races by SEM.     

Ultrastructural observations on the sensory papillae of juvenile and adult Gorgoderina vitelliloba (Trematoda: Gorgoderidae)

Hoole D. and Mitchell J.B. 1981. Ultrastructural observations on the sensory papillae of juvenile and adult Gorgoderina vitelliloba (Trematoda : Gorgoderidae). International Journal for Parasitology11: 411–417. Ultrastructural observations have been made on the juvenile and adult stages of Gorgoderina vitelliloba from Rana temporaria. Four types of sensory papillae occur; button, rosette, ciliated and domed. Button papillae, which contain a ciliary-rootlet but lack a cilium, occur on the oral sucker, dorsal preacetabular surface and the lateral margins of the fluke. Scanning electron microscopical observations reveal that the tegumental protuberances of the papilla are transformed from a spiked or conical appearance in juvenile flukes to a rounded form in adult flukes. Rosette papillae, which also contain a rootlet, occur on the lip of the ventral sucker of both juvenile and adult flukes. Ciliated papillae only occur on the oral sucker of juvenile flukes. Domed papillae, which contain a large area of electron-dense material, occur on the internal surface of the ventral sucker of both juvenile and adult flukes. The functions of papillae and their possible role in the migration of the parasite are discussed.     

Addendum to the Morphology of Steinernema scapterisci

This paper provides additional information on the morphology of Steinernema scapterisci. For females, the amphids are shown for the first time, and clearer scanning electron microscope (SEM) photographs of the six labial and four cephalic papillae are presented. For males, six labial and four cephalic papillae, one or two pairs of additional genital papillae, and the death shape are shown. For infective juveniles, six labial and four cephalic papillae and an elevated oral disc are shown.     

<Emphasis Type="Italic">Myrmeconema neotropicum</Emphasis> n. g., n. sp., a new tetradonematid nematode parasitising South American populations of <Emphasis Type="Italic">Cephalotes atratus</Emphasis> (Hymenoptera: Formicidae), with the discovery of an apparent parasite-induced host morph

A new genus and species of tetradonematid nematode, Myrmeconema neotropicum n. g., n. sp., is described from larval, pupal and adult stages of Cephalotes atratus L. (Hymenoptera: Formicidae) in Peru and Panama. Diagnostic characters of the new genus include: males and females subequal in size; cuticle with minute annulations; six cephalic papillae; stylet present in all stages; stichocytes absent; trophosome degenerate; three penetration glands; gonads paired and opposite; vulva in mid-body region; single spicule; genital papillae absent; adult tails rounded; infective juveniles moult once in egg; and adults of both sexes remain in the host throughout their development. As the female nematodes mature inside the worker ants, the host gasters change colour from black to red.     

Emodepside targets SLO-1 channels of Onchocerca ochengi and induces broad anthelmintic effects in a bovine model of onchocerciasis

Onchocerciasis (river blindness), caused by the filarial worm Onchocerca volvulus, is a neglected tropical disease mostly affecting sub-Saharan Africa and is responsible for >1.3 million years lived with disability. Current control relies almost entirely on ivermectin, which suppresses symptoms caused by the first-stage larvae (microfilariae) but does not kill the long-lived adults. Here, we evaluated emodepside, a semi-synthetic cyclooctadepsipeptide registered for deworming applications in companion animals, for activity against adult filariae (i.e., as a macrofilaricide). We demonstrate the equivalence of emodepside activity on SLO-1 potassium channels in Onchocerca volvulus and Onchocerca ochengi, its sister species from cattle. Evaluation of emodepside in cattle as single or 7-day treatments at two doses (0.15 and 0.75 mg/kg) revealed rapid activity against microfilariae, prolonged suppression of female worm fecundity, and macrofilaricidal effects by 18 months post treatment. The drug was well tolerated, causing only transiently increased blood glucose. Female adult worms were mostly paralyzed; however, some retained metabolic activity even in the multiple high-dose group. These data support ongoing clinical development of emodepside to treat river blindness.     

Dipetalonema viteae: Extraction and immunogenicity of cuticular antigens from female worms

The surface of adult female Dipetalonema viteae filarial worms was labeled with ¹²⁵I using 1,3,4,6-tetrachloro-3α, 6α-diphenylglycoluril as an iodinating reagent. Ultrastructural autoradiographs showed specific labeling of only the outermost cuticular layers. Surface-labeled worms were homogenized and were extracted with phosphate-buffered saline followed by NaOH. These extracts contained only minor amounts of radioactive TCA-precipitable material. Proteolytic digestion of the remaining sediment either with proteinase K, or thermolysin or subtilisin solubilized about 60% of the radioactive material of which 30% was TCA precipitable. Golden hamsters were injected with proteolytic extracts of unlabeled female worms. Subtilisin and thermolysin extracts provoked antibodies against somatic structures (e.g., gut, uterus, muscles) but not against the cuticle, whereas immunization with the proteinase K extract induced antibodies exclusively against cuticular and hypodermic structures of female and male worms.     

Repurposing Auranofin as a Lead Candidate for Treatment of Lymphatic Filariasis and Onchocerciasis

Two major human diseases caused by filariid nematodes are onchocerciasis, or river blindness, and lymphatic filariasis, which can lead to elephantiasis. The drugs ivermectin, diethylcarbamazine (DEC), and albendazole are used in control programs for these diseases, but are mainly effective against the microfilarial stage and have minimal or no effect on adult worms. Adult Onchocerca volvulus and Brugia malayi worms (macrofilariae) can live for up to 15 years, reproducing and allowing the infection to persist in a population. Therefore, to support control or elimination of these two diseases, effective macrofilaricidal drugs are necessary, in addition to current drugs. In an effort to identify macrofilaricidal drugs, we screened an FDA-approved library with adult worms of Brugia spp. and Onchocerca ochengi, third-stage larvae (L3s) of Onchocerca volvulus, and the microfilariae of both O. ochengi and Loa loa. We found that auranofin, a gold-containing drug used for rheumatoid arthritis, was effective in vitro in killing both Brugia spp. and O. ochengi adult worms and in inhibiting the molting of L3s of O. volvulus with IC₅₀ values in the low micromolar to nanomolar range. Auranofin had an approximately 43-fold higher IC₅₀ against the microfilariae of L. loa compared with the IC₅₀ for adult female O. ochengi, which may be beneficial if used in areas where Onchocerca and Brugia are co-endemic with L. loa, to prevent severe adverse reactions to the drug-induced death of L. loa microfilariae. Further testing indicated that auranofin is also effective in reducing Brugia adult worm burden in infected gerbils and that auranofin may be targeting the thioredoxin reductase in this nematode.     

Margolisianum bulbosum n. gen., n. sp. (Nematoda: Philometridae) from the southern flounder, Paralichthys lethostigma (Pisces: Bothidae), in Mississippi Sound, USA

A new species of a philometrid nematode, Margolisianum bulbosum, is described from the subcutaneous tissue in the mouth (larvigerous females), head (males, ovigerous, and larvigerous females), and eye (preovigerous females) of the southern flounder, Paralichthys lethostigma, from Mississippi Sound. It is placed in a new genus diagnosed by the combination of 8 large, paired but separate cephalic papillae; no inner cephalic papillae; an esophagus with a separate, muscular anterior bulb; a prominent mononuclear esophageal gland; and variable, irregularly distributed cuticular bosses in the females, as well as a vestigial rectum, particularly in larvigerous females. Some female specimens exhibit rows of lateral grooves and longitudinal ridges near the posterior end. Males have 2 small slightly subequal spicules, a barbed gubernaculum, 4 pairs of small cephalic papillae, and a bipartite hypodermal extension within a membranous cuticle on the posterior end. Males, ovigerous females, and larvigerous females appear to be present year round in this sporadic infection in Mississippi.     

Description of Boleodorus Bushehrensis n. Sp. (Rhabditida: Tylenchidae) from Southern Iran,and Observations on a Commonly Known Species

A new species of the genus Boleodorus, recovered from southern Iran, is described and illustrated based upon morphological and molecular data. B. bushehrensis n. sp. is mainly characterized by having a wide and low cephalic region (which is continuous with the adjacent body), the oral aperture in a depression in side view under a light microscope, four lines in the lateral field, weak metacorpus with a vestigial-to-invisible valve, and short (26–38 mm long) hooked tail with rounded tip. The females are 334–464 mm long and have a spherical spermatheca filled with spheroid sperm; males have 11.5- to 12.0-mm-long spicules and weakly developed bursa. The new species has an annulated low cephalic region, four large cephalic papillae, and small crescent-shaped amphidial openings when observed by scanning electron microscopy (SEM). Its morphological and morphometric differences with seven known species are discussed. The phylogenetic relationships of the new species with other relevant genera and species have been studied using partial sequences of small and large subunit ribosomal DNA (SSU and LSU rDNA). In both the SSU and LSU phylogenies, the sequences of B. bushehrensis n. sp. and other Boleodorus spp. formed a clade. A second species, B. thylactus, when studied under SEM, has a raised, smooth cephalic region, four large cephalic papillae, and oblique amphidial slits, with the oral opening in a depression.     

Host-induced effects on the morphology of Ostertagia circumcincta in grazing sheep

Waller P. J. and Thomas R. J. 1978. Host-induced effects on the morphology of Ostertagia circumcincta in grazing sheep. International Journal for Parasitology8: 365–370. Morphological changes are described in adult Ostertagia circumcincta recovered from lambs slaughtered monthly from June to November. Comparisons were made between continuously grazed principal lambs exposed to infection for increasingly long periods, and tracer Iambs exposed for only four weeks. Lambs were either slaughtered immediately off pasture or held 2 weeks under worm free conditions before slaughter. Although no resistance to establishment of infection occurred, shown by comparable worm burdens of principals and tracers, a rapid parasite population turnover was observed in the principal lambs which ceased once the lambs were removed from the infected pasture. Significantly fewer adult female Ostertagia with fully developed vulval flaps were recorded in principals compared to tracers. Such differences are considered to be primarily host induced responses which rapidly come into play, evidenced by the entire populations of developing female worms in tracers slaughtered immediately off pasture showing arrested vulval flap development after they had completed development in those tracers held for a further two weeks. A progressive reduction in the proportion of adult male worms also occurred in the principal lambs. Population density was shown to have an inverse correlation with worm size. By analogy, it is suggested that seasonal change in the presence of cuticular inflations in Haemonchus contortus females with linguiform vulval flaps may be primarily host induced, rather than a change in the relative fitness of discrete genotypes.