Identification of carbohydrates on the surface membrane of pathogenic and nonpathogenic piscine haemoflagellates, Cryptobia salmositica, C. bullocki and C. catostomi (Kinetoplastida)

Feng, Shujuan; Woo, P. T. K.

doi:10.3354/dao032201

Cited by 8 publications

(6 citation statements)

References 20 publications

(25 reference statements)

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“…The intense 108 kDa band observed in R and in C blots apparently belongs to the host in accordance with the absence of NeuAc residues previously re ported for E. leei and E. scophthalmi by immunohistochemistry (Redondo et al 2008, Redondo & Álvarez-Pellitero 2009) as well as for Myxobolus cerebralis by lectin blot (Kaltner et al 2007). However, surface exposed sialic acids may play a role in self/non-self recognition, and therefore some parasites would acquire sialic acid moieties from the host by trans-sialidases as a biological mask to evade immune detection by phagocytes or complement (Feng & Woo 1998b, Kaltner et al 2007). Therefore, this band might be alternatively interpreted as a sialylated glycoconjugate of the parasite.…”

Section: Discussionmentioning

confidence: 98%

“…In fish, as in all vertebrates, the lectin pathway of the complement system is an ancient first line defense mechanism of the innate immune system that relies on recognition of pathogen-associated glycan epitopes (Sunyer & Lambris 1998, Nakao et al 2006, Kania et al 2010. In previous studies, the glycoproteins of some piscine parasites have been studied by lectin blotting (Feng & Woo 1998b, Kim et al 1999, Muñoz et al 2000a, Knaus & El-Matbouli 2005.…”

Section: Introductionmentioning

confidence: 99%

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Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea)

Estensoro¹,

Álvarez-Pellitero²,

Sitjà‐Bobadilla³

2013

Dis. Aquat. Org.

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Enteromyxum leei, an intestinal myxozoan parasite affecting a wide range of fish, was partially purified, and the immunogenic composition of its glycoproteins as well as the proteolytic activity were studied. Parasite extracts, mainly containing spores, were separated by SDS-PAGE, and thereafter, immunoblots were carried out with a polyclonal antiserum (Pab) raised against E. leei. Periodic acid/Schiff staining of gels, periodate- and Proteinase K-treated Western blots and Lectin blots were performed to analyse the terminal carbohydrate composition of the parasite's antigens. Additionally, the cross-reaction of the parasite extracts with a Pab raised against the polar filament of the myxozoan Myxobolus pendula was studied. Both Pabs detected proteic epitopes on antigenic proteins and glycoproteins of E. leei, ranging between 15 and 280 kDa. In particular, 2 glycoproteic bands (15 and 165 kDa), immunoreactive to both Pabs and with glucose and mannose moieties, could correspond to common antigens shared among myxozoans. The 165 kDa band also presented galactose, N-acetyl-galactosamine and N-acetyl-glucosamine, pointing to its possible origin on chitin-built spore valves and to its possible involvement in host-parasite interactions. The molecular weight of the 15 kDa glycoproteic antigen matches that of minicollagen, a cnidarian-specific protein of nematocysts with a myxozoan homologue. Several proteases with apparent molecular weights ranging between 43 and 245 kDa were found in zymographies of E. leei extracts, and these may have a potential role in the parasite's pathogenesis. This is a useful approach for further studies to detect targets for antiparasitic therapy.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea)

Estensoro¹,

Álvarez-Pellitero²,

Sitjà‐Bobadilla³

2013

Dis. Aquat. Org.

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“…There are also differences in expression of surface carbohydrate residues and glycoproteins on cell membranes of the pathogenic and vaccine strains; specifically, the increased surface carbohydrate residues coincide with its loss of virulence (Feng & Woo 2001). In another study Feng & Woo (1998d) showed the non‐pathogenic Cryptobia catostomi had more carbohydrates on its surface and had stronger reactions with lectins than the pathogenic C. salmositica .…”

Section: Glycoproteins Surface Membrane and Metabolismmentioning

confidence: 95%

“…It has been cloned and its biology studied (e.g. Woo & Thomas 1991, 1992; Zuo & Woo 1996, 1997a; Feng & Woo 1998d, 2001). A single intraperitoneal injection of the live vaccine protects 100% of fish (juveniles or adults) from disease and mortality.…”

Section: Acquired Immunitymentioning

confidence: 99%

Cryptobia (Trypanoplasma) salmositica and salmonid cryptobiosis

Woo

2003

Journal of Fish Diseases

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Salmonid cryptobiosis is caused by Cryptobia (Trypanoplasma) salmositica. The haemoflagellate has been reported from all species of Pacific Oncorhynchus spp. on the west coast of North America. It is normally transmitted by the freshwater leech, Piscicola salmositica, in streams and rivers, and sculpins, Cottus spp., are considered important reservoir hosts. The pathogen can also survive on the body surface of fish because it has a contractile vacuole to osmoregulate when the fish is in fresh water. This allows for direct transmission between fish, especially in aquaculture facilities. The parasite divides rapidly by binary fission in the blood to cause disease, the severity of which is directly related to parasitaemia. Cryptobia salmositica has a mitochondrium and it normally undergoes aerobic respiration; however, if its mitochondrium is damaged it will switch to glycolysis. Its glycolytic enzymes and catalase are contained in glycosomes. Cysteine protease is a metabolic enzyme, and its neutralization inhibits oxygen consumption and multiplication of the parasite. An important virulent factor in cryptobiosis is a secretory metalloprotease. The protective mechanism involves production of complement fixing antibodies, phagocytosis by macrophages, and cell-mediated cytotoxicity. Recovered fish are protected, probably for life as the immunity is non-sterile. Clinical signs of the disease include anaemia, anorexia, splenomegaly, general oedema and abdominal distension with ascites. The metabolism and swimming performance of infected fish are significantly reduced and the bioenergetic cost of the disease is very considerable. Fish are susceptible to hypoxia and their immune system is depressed during acute cryptobiosis. Severity of the disease and mortality rates vary significantly between species and stocks of salmon. Protective strategies include selective breeding of Cryptobia-resistant fish. This is innate resistance to infection and it is controlled by a dominant Mendelian locus. In these fish the parasite is lysed via the alternative pathway of complement activation. In Cryptobia-tolerant fish (infected with the pathogen but which do not suffer from disease) the metalloprotease secreted by the parasite is neutralized by alpha2 macroglobulin. Hence, the production of a transgenic Cryptobia-tolerant salmon is an option. This strategy has the advantage in that human intervention (e.g. vaccination, chemotherapy) is not required once the transgenic fish is produced. Acquired immunity is another option; a single dose of the attenuated live vaccine protects fish for at least 2 years. The protective mechanism in vaccinated fish is similar to that in recovered fish. The trypanocidal drug, isometamidium chloride, is an effective therapeutic and prophylactic agent. It accumulates in the mitochondrium of the parasite and significantly disrupts aerobic respiration by causing lesions in the organelle. Efficacy of the drug is significantly increased after its conjugation to antibodies. This immuno-chemotherapeutic strategy ha...

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“…In aquatic parasites, surface associated glycans were identified in Tetracapsuloides bryosalmonae (Morris and Adams 2004), Eudiplozoon nipponicum (Schabussova et al 2004), in monogenean parasites (Buchmann and Lindenstrom 2002) and in Cryptobia spp. (Feng and Woo 1998). The interaction of such glycans with lectins, including congerins I and II (Muramoto and Kamiya 1992;Muramoto et al 1999) and AJL 1 and 2 (Tasumi et al 2002(Tasumi et al , 2004 in fish mucous, may constitute a recognition system involved in the attachment of ichthyoparasites to the epidermis of the host.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of carbohydrate-binding sites in developmental stages of Myxobolus cerebralis

Knaus¹,

El‐Matbouli²

2005

Parasitol Res

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Glycans and lectins (carbohydrate-binding molecules) form a mutual recognition system, which enables parasitic organisms to attach themselves to the host cells and/or take part in the migration of their developmental stages into the target tissue. The aim of the present study was to identify and characterise the potential binding activity of glycoconjugates in different developmental stages of Myxobolus cerebralis, the causative agent of whirling disease in salmonids. The binding patterns of 13 biotinylated neoglycoconjugates were histochemically examined in thin-sections of infected rainbow trout (Oncorhynchus mykiss) and oligochaetes (Tubifex tubifex), as well as isolated waterborne triactinomyxon spores. A distinct structure-selective and developmental stage-regulated expression of certain classes of carbohydrate binding was observed. In triactinomyxon spores, the expression of carbohydrate binding activity for alpha-l-Fuc-BSA-biotin, alpha-d-GalNAc-BSA-biotin, beta-d-GlcNAc-BSA-biotin, Lac-BSA-biotin and ASF-biotin was up-regulated in the polar capsules; the shell valves showed no activity. In the gut of T. tubifex, polar capsules of the parasite showed strong positive reaction only for beta-d-GlcNAc-BSA-biotin. In fish cartilage, polar capsules were negative, but the spore shell valves showed a broad range of carbohydrate binding activity. No activity was detected for either alpha6- or alpha3-linked N-acetyl-d-neuraminic acid to galactose. An adhesion assay was performed on GlycoWell plates and Myxobolus spores were found to specifically adhere to matrices containing residues of lactose, fucose, galactose, N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. This is the first study to identify lectin activity in a myxozoan parasite; activity that is likely to play a role in the recognition systems involved in host specificity and the processes of spore attachment and invasion.

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Identification of carbohydrates on the surface membrane of pathogenic and nonpathogenic piscine haemoflagellates, Cryptobia salmositica, C. bullocki and C. catostomi (Kinetoplastida)

Cited by 8 publications

References 20 publications

Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea)

Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea)

Cryptobia (Trypanoplasma) salmositica and salmonid cryptobiosis

Characterisation of carbohydrate-binding sites in developmental stages of Myxobolus cerebralis

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