Changes in the level of acetylcholinesterase of Nematospiroides dubius and Trichostrongylus colubriformis following paralysis by levamisole in vivo

Acetylcholinesterase (acetylcholine acetylhydrolase; EC 3.1.1.7) levels of Nematospiroides dubius from laboratory mice and Trichostrongylus colubriformis from lambs have been measured. The anthelmintic levamisole (leavo isomer of 2,3,5,6-tetrahydro-6-phenylimidazo-(2,1b)-thiazole (Tetramizole)) did not affect the level of acetylcholinesterase in N. dubius in vivo but caused a reduction in the level of the enzyme in T. colubriformis following paralysis in vivo. The effect of levamisole on acetylcholinesterase in the nematodes is explained in terms of the differing roles of the enzyme in these two species.     

鱼类寄生宫脂属线虫研究进展

宫脂属线虫是鱼类寄生线虫的重要类群。本文从分类学、流行病学、生态学以及生活史等方面对宫脂属线虫的研究进展进行了论述。     

Interaction of ivermectin with γ-aminobutyric acid receptors in Trichinella spiralis muscle larvae

The value of the γ-aminobutyric acid (GABA) receptor of nematodes as a target for ivermectin's mode of action remains unclear. Using binding assays, we examined extracts from Trichinella spiralis muscle larvae for the presence of [³H]-ivermectin and [³H]-GABA binding sites. Tissue preparations displayed affinity binding sites for [³H]-ivermectin with a dissociation constant (K _d) of 83 nM and a receptor density (B_max) of 145 fmol/mg protein. We also identified a specific [³H]-GABA binding activity with a K _d of 1.2 μM and a B_max of 4.78 pmol/mg protein. In competition studies, ivermectin was found to be a competitive inhibitor of specific [³H]-GABA binding activity with an inhibition constant (K _i) of 3.39 nM, suggesting that GABA receptors could be implicated in the mechanism of action of ivermectin in nematodes. Received: 7 August 1998 / Accepted: 22 September 1998     

Strategic use of moxidectin or closantel in combination with levamisole in the control of nematodes of sheep in the highlands of central Kenya

The strategic use of moxidectin or closantel in combination with levamisole (LEV) to control gastrointestinal nematodes of sheep in the highlands of central Kenya was examined. Thirty Corriedale female lambs aged between 6 and 8 months were assigned to three treatment groups of ten lambs each. The three groups of lambs were set stocked on separate paddocks for the entire study period of 12 months. Lambs in Group 1 were dewormed strategically with moxidectin at 0.2 mg/kg body weight and those in Group 2 with closantel at 10 mg/kg body weight together with LEV at 7.5 mg/kg body weight. These strategic treatments were given 3 weeks after the onset of both the short and long rains and at the end of the long rainy season. The third group of lambs remained untreated (control group). Nematode infections in the treated groups of lambs and larval infectivity for the pastures on which the lambs were grazing were well controlled compared with the untreated control group. This resulted in higher weight gains and packed cell volume (PCV) in the treated lambs compared with the untreated lambs. These parameters were comparable between the lambs treated with moxidectin and those treated with closantel plus LEV. The estimated monitory benefit per animal from the control of gastrointestinal nematodes using moxidectin or closantel in combination with LEV when compared with animals in the control group were US dollars 26 and 25, respectively. It was concluded that worm control strategies for sheep in the study area, which are based on anthelmintic treatments during the rainy seasons, are effective. Due to the extended period during which pastures remain infective in the high rainfall central highlands of Kenya, anthelmintics with sustained action such as moxidectin or closantel may be most effective. On farms where resistance to the commonly used benzimidazoles or LEV groups of anthelmintics has developed, moxidectin or closantel may be used in helminth control programs for sheep.     

Genus-specific PCR for the differentiation of eggs or larvae from gastrointestinal nematodes of ruminants

Using sequences of the ribosomal second internal transcribed spacer (ITS2), PCR primers were designed for the differentiation of the gastrointestinal nematode genera Ostertagia, Cooperia, Nematodirus, Haemonchus and Trichostrongylus. Single eggs or larvae from faeces could be differentiated without previous DNA extraction. Quantification of the PCR result proved to be difficult because the DNA content between eggs from fresh or 24-h-old faeces varied considerably. Received: 12 February 1999 / Accepted: 2 May 1999     

Dolichol kinase in Ascaris suum and Onchocerca volvulus

Dolichol kinase was demonstrated in the microsomal fraction of Ascaris suum and Onchocerca volvulus. The enzyme from nematodes exhibited specificity for CTP as phosphoryl donor and was found to be inhibited by the reaction product CDP. Enzyme activity was optimal at pH 7.4, in the temperature range between 30 degrees and 37 degrees C, and in the presence of 0.5% Triton X-100. In addition, the enzyme was found to depend on divalent cations for activity; Mg2+ being more effective than Mn2+ and Ca2+. The dolichol kinase from both nematodes was shown to be independent of Ca2+-calmodulin for activity. The apparent Km values for dolichol were determined to be 7.5 and 9.0 micrograms ml-1 for the enzyme from A. suum and O. volvulus, respectively. Those for CTP were estimated to be 0.85 and 0.75 mM, respectively.     

Cyclic AMP-dependent and independent protein kinases in Ascaridia galli

The occurrence of multiple protein kinases, distinguished with respect to molecular weight and preference for acceptor proteins, was demonstrated in Ascaridia galli. The molecular weights of the cyclic AMP-dependent protein kinase and of phosvitin kinase I and II — both independent of cyclic AMP — were determined to be 160 000, > 200 000 and 40 000, respectively. The cyclic AMP-dependent protein kinase preferred histones and kemptide as acceptor substrates; stimulation of enzyme activity was up to 4-fold by cyclic AMP. The activities of phosvitin kinase I and II were found to be effectively inhibited by suramin. The inhibition constants were calculated to be 2 μM and 5 μM, respectively. In addition, stibophen turned out to be a potent inhibitor of phosvitin kinase I; the inhibition constant was determined to be 10 μM.     

Cuticular surface lipids are responsible for disguise properties of an entomoparasite against host cellular responses

Entomopathogenic nematodes are widely used as alternatives to chemicals for the biological control of insects. These endoparasites are symbiotically associated with bacteria that are lethal for the host; however, parasites need to overcome the host immune defences to complete a successful life cycle. The processes parasites employ to escape or depress host immunity are targeted at deceiving non-self recognition as well as inactivating defence reactions.

The purpose of this paper is to investigate the interactions between the entomopathogenic nematode Steinernema feltiae and the lepidopteran Galleria mellonella, focusing on the role of the parasite's body-surface compounds in the immunoevasion of host cell-mediated responses.

To evaluate host self/non-self discrimination and encapsulation efficiency, we carried out a series of interaction assays between cultured host hemocytes and parasites or isolated cuticles. The data obtained suggest that the parasite cuticular lipids (PCLs) are able to bind a variety of host hemolymph molecules; PCLs attract host proteins from the hemolymph creating a coat around the parasite, thus, enabling Steinernema to disguise itself against hemocytes recognition. The role of parasite lipids in the disguise process was also investigated by simulating the nematode body surface with agarose microbeads covered with purified cuticular components; when the beads were coated with cuticular lipids, host hemocytes were not able to recognize and encapsulate.Results suggest that by means of attracting host hemolymph components onto its cuticular surface, S. feltiae prevents hemocytes attachment to its cuticle and inhibits melanization by depleting hemolymph components.