Fasciola gigantica: Ultrastructural cytochemistry of the tegumental surface in newly- excysted metacercariae and in vitro-penetrated juvenile flukes informs a concept of parasite defence at the interface with the host

The subcellular route of incorporation of complex carbohydrates into rabbit heterophil primary granules and their subsequent intragranular distribution during granule maturation were studied with ultrastructural, cytochemical, and radioautographic methods. High iron diamine (HID) staining of sulfated glycoconjugates in primary granules was partially diminished after treatment with chondroitinase ABC or after removal of N-sulfate groups with nitrous acid, but was not altered by exposure to hyaluronidase, trypsin, or HCl. Subsequent thiocarbohydrazide-silver proteinate (TCH-SP) straining of thin sections increased the density of the HID reaction product. Golgi-derived spherules and very immature morular granules stained weakly with HID-TCH-SP and labeled intensely after a 10 min incubation with 35SO4. After a 60 min 35SO4 pulse and a 60 min chase, an increase in radiolabeling was observed in granules with HID stained, fused morular material, and some labeling was present in more mature rim stained granules. Fully mature granules lacked HID or HID-TCH-SP staining, but contained most of the 35SO4 labels after a 60 min pulse and 18 hr chase in vitro. Periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of unosmicated thin sections localized vicinal glycol-containing complex carbohydrates in Golgi-associated small vesicles. These vesicles lacked HID-TCH-SP staining and apparently contained neutral glycoprotein. They frequently bordered, in a rosette arrangement, the immature morular granules, but not the more mature primary granules. The PA-TCH-SP method localized complex carbohydrates in the rim of granules precursors and enclosed a spherule or morula, but failed to stain the sulfate-containing material in the morulas or spherules. PA-TCH-SP reactivity was diffusely distributed in moderately mature granules and was decreased in fully mature granules. These results indicate that heterophil primary granule contain several complex carbohydrates including O-sulfated and N-sulfated glycosaminoglycans, as well as vicinal glycol-containing glycoproteins. These complex carbohydrates are transported to immature primary granules by different Golgi-derived organelles. The complex carbohydrates are subsequently distributed differently within primary granules and become masked to staining as the granule matures.

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In vitro tegumental alterations on adult Fasciola gigantica caused by mefloquine

Journal of Parasitic Diseases ◽

10.1007/s12639-014-0466-y ◽

2014 ◽

Vol 40 (1) ◽

pp. 145-151 ◽

Cited By ~ 2

Author(s):

Hatem A. Shalaby ◽

Amira H. El Namaky ◽

Reem O. A. Kamel

Keyword(s):

Fasciola Gigantica

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The anthelmintic activity of 1,10-phenanthroline, a metalloenzyme inhibitor

Indian Journal of Animal Research ◽

10.18805/ijar.b-3692 ◽

2019 ◽

Author(s):

S.M.A. Abidi ◽

Kavita Singh ◽

A. Rehman ◽

R. Ullah ◽

L. Rehman ◽

...

Keyword(s):

Enzyme Inhibitor ◽

Anthelmintic Activity ◽

In Vivo Studies ◽

Parasitic Helminths ◽

Tegumental Surface ◽

Biochemical Assays ◽

Direct Exposure ◽

Surface Changes

Paramphistomosis is a chronic, debilitating parasitic disease of livestock prevalent in the tropical and sub-tropical countries. Globally there is a heavy reliance on anthelmintics but concerns over drug resistance encourage the search for new leads. Metalloproteinases play a significant role in the biology and life cycle of parasitic helminths. The efficacy of metalloproteinase inhibitor, 1,10-Phenanthroline (1,10-phe) which is commonly used as a specific enzyme inhibitor in biochemical assays, was tested in vitro on Gigantocotyle explanatum tegument as a marker of anthelmintic action. The scanning electron microscopy revealed that the tegumental surface exhibited considerable changes in the worms treated with the metalloenzyme inhibitor, 1,10-phe. The untreated control worms appeared normal showing smooth tegumental surface with abundant dome shaped papillae in the anterior to mid region, while their density was less around the acetabulum which serves as a hold-fast organ helping the worms to remain attached in biliary passage. The 1,10-phe produced significant tegumental damage when the liver amphistomes were in vitro exposed to this compound at 12.5 µM concentration. The surface changes appeared in the form of edematous ridges with prominent furrows and erosion of the dome shaped papillae with rosette shaped deep lesions as a result of which deep parenchymatous tissues were exposed. The collapse of sensory bulbs as well as sloughing of tegument, particularly in the anterior-mid region was observed. The nature of damage could be comparable to various anthelmintics used in previous studies. To the best of our knowledge this is the first report of direct exposure of amphistome worms to zinc metallo-enzyme inhibitor, however, further in vivo studies are required to ascertain the anthelmintic efficacy of 1,10-phe.

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Gamal (Gliricidia Sepium) Leaf Extract Increase Mortality of Fasciola gigantica In Vitro

Jurnal Medik Veteriner ◽

10.20473/jmv.vol4.iss2.2021.193-198 ◽

2021 ◽

Vol 4 (2) ◽

pp. 193

Author(s):

Brillia Zulianti ◽

Iwan Sahrial Hamid ◽

Aditya Yudhana ◽

Jola Rahmahani ◽

Maya Nurwartanti Yunita ◽

...

Keyword(s):

Mortality Rate ◽

Leaf Extract ◽

In Vitro Study ◽

Gliricidia Sepium ◽

Fasciola Gigantica ◽

Economic Losses ◽

Randomized Design ◽

The Mean ◽

Range Test

Fasciolosis is a disease caused by infection with liver fluke (Fasciola sp). Fasciolosis is a parasitic disease that is important because it can cause high economic losses. The aim of this study was to investigate the effect of gamal (Gliricidia sepium) leaf extract against mortality of Fasciola gigantica in vitro study. In present study was completely randomized design with six treatments and each treatment were divided into four replications. A total of 240 specimens from infected cattles were used in this study. The observation and recording of F. gigantica mortality in all treatments with concentrations of gamal leaf extract P1 (1%), P2 (2%), P3 (3), P4 (4%), K + (Nitroxynil 2%) and K- (physiological NaCl) rate was done in 30, 60, 90, 120, 150, 180, 210 and 240 minutes, respectively. The mortality of F. gigantica was confirmed if there was no movement when triggered by anatomy tweezers then dipped in slightly warm water (50 15â„ƒ"> ). The data was analyzed using ANAVA Faktorial and continued with Duncan Multiple Range Test. The results of this study were shown the lowest and the highest mortality rate recorded at minutes 30 and 240 respectively. Moreover, the mean of mortality rate were peaked in nitroxynil (K+) and 1% gamal leaf extract treatment, while the lowest mean of mortality rate occurs in NaCl (K-) treatment. Present study conclude that gamal leaf extract has anthelmintic effect on the mortality rate of Fasciola gigantica. Gamal leaf extract with 1% concentration was claimed to be the most effective.

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Modulation of the Functions of Goat Peripheral Blood Mononuclear Cells by Fasciola gigantica Thioredoxin Peroxidase In Vitro

Pathogens ◽

10.3390/pathogens9090758 ◽

2020 ◽

Vol 9 (9) ◽

pp. 758 ◽

Cited By ~ 1

Author(s):

Ai-Ling Tian ◽

Xiaowei Tian ◽

Dan Chen ◽

Mingmin Lu ◽

Guillermo Calderón-Mantilla ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Transforming Growth Factor ◽

Interleukin 2 ◽

Fasciola Gigantica ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Thioredoxin Peroxidase

The liver fluke Fasciola gigantica has a remarkable ability to establish a long-term infection within the hepatobiliary system of the mammalian definitive host. F. gigantica achieves this by producing excretory–secretory molecules, which have immunomodulatory activities. In an effort to elucidate the immunomodulatory functions of F. gigantica thioredoxin peroxidase protein (FgTPx), we expressed recombinant FgTPx (rFgTPx) in Escherichia coli bacteria and examined its effects on several functions of goat peripheral blood mononuclear cells (PBMCs) in vitro. Sequence analysis revealed that FgTPx is related to a thioredoxin-like superfamily. Western blot analysis showed that rFgTPx was recognized by the sera of goats experimentally infected by F. gigantica. The specific binding of rFgTPx protein to the surface of goat PBMCs was demonstrated by immunofluorescence staining. We investigated the influence of serial concentrations of rFgTPx on various functions of goat PBMCs. All concentrations of rFgTPx increased the secretion of interleukin-2 (IL-2), IL-4, IL-10, IL-17, transforming growth factor-beta (TGF-β), and interferon gamma (IFN-γ), but inhibited PBMC proliferation, migration, and monocyte phagocytosis. Goat PBMCs exposed to 20–40 μg/mL of rFgTPx secreted increased levels of nitric oxide (NO), and 10–40 μg/mL of rFgTPx promoted cell apoptosis. These findings indicate that rFgTPx influences various functions of goat PBMCs by interacting with a large number of cellular targets, ultimately to promote the parasite’s survival. The roles of rFgTPx and their interacting proteins warrant further investigation.

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