Use of fluorescent lectin binding to distinguish Teladorsagia circumcincta and Haemonchus contortus eggs, third-stage larvae and adult worms

Abstract

Lectin binding to carbohydrates on parasite surfaces has been investigated as a method of distinguishing adult worms, eggs and sheathed and exsheathed L3 of Teladorsagia circumcincta and Haemonchus contortus, economically important abomasal parasites in temperate climates. Both species were maintained as pure laboratory cultures of field isolates from New Zealand. Each of the four life cycle stages could be distinguished by the binding of at least one lectin: adult worms by Sambucus nigra agglutinin (SNA); eggs by peanut agglutinin (PNA), ConcavalinA and Lens culinaris agglutinin (LCA); exsheathed L3 by Griffonia simplicifolia-I lectin (GSL-I) and Lotus tetragonolobus lectin (LTL) and sheathed L3 by Aleuria aurantia lectin (AAL). The whole surface of both adult T. circumcincta and H. contortus strongly bound lectins specific for N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), mannose and fucose, but the two species could be distinguished by SNA binding only to T. circumcincta. Eggs could be distinguished by the binding of mannose-specific PNA to H. contortus and GalNAc-specific LCA and PSA to T. circumcincta eggs. GalNAc, GlcNAc and mannose lectins bound to the cuticle and over the excretory pores of a large proportion of sheathed L3 of both species, but only the H. contortus surface had exposed fucose or sialic acid complexes. The distinguishing lectin for sheathed L3 was AAL, which did not bind to T. circumcincta, but bound weakly to the head region of all fresh H. contortus and to 50-90% after 3 months storage. The cuticle of exsheathed L3 was unresponsive to all 19 lectins, and any binding was restricted to the head and tail regions. L3 exsheathed after 2-4 months storage could be distinguished by the binding of GSL-I and LTL to H. contortus but not to T. circumcincta. Lectin binding could be a useful adjunct in identifying L3, but lacked the consistency to be definitive, whereas it could be further developed as a practical method of distinguishing parasitic nematodes at other stages in the life cycle, particularly the eggs.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus

“…Therefore, the ricin protein, a toxic protein with 60 kDa which is contained in R. communis seeds (WORBS et al, 2011), cannot be the active compound against H. contortus eggs, although it is known that the surface of H. contortus eggshells contains galactose carbohydrate (PALMER; McCOMBE, 1996;JURASEK et al, 2010;HILLRICHS et al, 2012), which is a binding site for ricin. Trypsin inhibitor activity, albeit weak, was also observed in the CERN fraction of R. communis.…”

“…Many studies have shown the presence of carbohydrates on GIN surfaces, using labeled lectins (BONE;BOTTJER, 1985;PALMER;McCOMBE, 1996;JURASEK et al, 2010;HILLRICHS et al, 2012). In these studies, Concanavalin A (ConA), Lens culinaris agglutinin (LCA), Maclura pomifera agglutinin (MPA), peanut agglutinin (PNA) and Pisum sativum agglutinin (PSA) were bound to the egg surfaces, suggesting the presence of glucose, mannose and galactose carbohydrates.…”

Targeting Aberrant Sialylation in Cancer Cells Using a Fluorinated Sialic Acid Analog Impairs Adhesion, Migration, and In Vivo Tumor Growth

“…a2,3-linked sialic acids were detected using MALII, a2,6-linked sialic acids by SNA-I, exposure of terminal (b-)galactose/T antigen by PNA, exposure of terminal (a-)galactose and (a-)GalNAc by GSL-I, (poly) GlcNAc (chitobiose) by WGA, and (a-)fucose by AAL (30)(31)(32). Free lectin was removed by washing the cells in carbofree blocking solution, and cell-bound biotinylated lectin was conjugated with 2 mg/mL streptavidin-phycoerythrin for 15 minutes (33).…”

Allosteric Regulation of the Carbohydrate-binding Ability of a Novel Conger Eel Galectin by d-Mannoside

“…V. anguillarum) via a galactose or lactose sugar unit, whereas Con-P recognizes oligo-mannose sugars via allosteric regulation. Actually, some mannose-specific lectins bind strongly to parasitic nematodes and nematode eggs (18). Furthermore, Con-P was detected and cloned from the peritoneal cells and the cells that encapsulated parasitic nematodes in the abdominal cavity (24), and Con-P interacted with yeast mannan (supplemental Fig.…”