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The metabolic profile of adult Fasciola hepatica obtained from rafoxanide-treated sheep.

Sheep infected with adult Fasciola hepatica were drenched with rafoxanide. At 4, 8, 16 and 24 h after drenching the sheep were killed and the flukes removed, washed and rapidly frozen in liquid nitrogen. The content of key metabolites in the fermentation pathway were determined and compared with those in control F. hepatica, whose hosts were not treated with rafoxanide. Rafoxanide decreased glycogen, malate, NADH and ATP levels. The level of other metabolites in the pathway increased for the first 8-16 h after rafoxanide treatment. The marked decrease in ATP and glycogen, and the increase in total [NAD+]/[NADH] and [oxaloacetate]/[malate], together with the changed content of other metabolites, led to the conclusion that the mode of action of rafoxanide against F. hepatica in vivo is by uncoupling oxidative phosphorylation.

Adenosine Diphosphate

Trials with rafoxanide. 8. Efficacy of an injectable solution against trematodes and nematodes in cattle.

Four experiments are described in which the efficacy of an experimental 5% injectable solution of rafoxanide was evaluated against various adult and immature helminths in cattle. Subcutaneous injection at a dosage of 3 mg/kg live mass resulted in the following reductions in mean worm burdens: adult Fasciola hepatica, 82,6%; adult Fasciola gigantica, 99,8% immature Paramphistomum microbothrium, 10,1% adult Haemonchus placei, 99,6%, third stage H. placei, 73,7%; adult Bunostomum phlebotomum, 99,8%; adult Oesophagostomum radiatum, 99,9%; and fourth stage O. radiatum, 76,9%. At 5 mg/kg live mass, rafoxanide solution was 97,5% and 99,2% effective against 8-week old F. gigantica and third stage H. placei respectively and at 7,5 mg/kg, 92,4% against 6-week old F. gigantica.

Animals

[Behavior of selected veterinary preparations during industrial milk processing].

Since the extensive veterinary-hygienic treatment of herds with insecticidal veterinary preparations may lead to milk contamination, the authors studied the behaviour of some active agents (butonate, dichlorvos (DDVP), trichlorphon, naled, carbaryl, hexachloro-p-xylene and rafoxanide) during the industrial processing of milk to fluid milk, cream, butter, cheese and milk powder, and during the storage of these products. Small-scale model experiments served to investigate the effects of pasteurization (74 and 95 degrees C.), separation and churning as well as of the processing to milk powder and cheese. Analyses for residues were performed by thin-layer chromatography, colorimetry and with the aid of the isotope technique. From the viewpoint of milk processing, the use of TCP and, in part, that of DDVP may be considered as less critical due to their hydrophilic properties and rapid degradation. In view of their lipophilic behaviour, the use of butonate, carbaryl, rafoxanide and hexachloro-p-xylene as active agents in veterinary preparations for milk cows must be regarded as problematic. The utilization of naled is also problematic due to the fact that the toxicology of its metabolites is not yet sufficiently known.

Animals

Effect of anthelmintic treatment on the performance of Awassi sheep in Iraq.

Studies are reported on the effect of anthelmintic treatment on the body weight gain and wool growth of ewes with subclinical helminth infections. The parasites involved were Fasciola gigantica and Ostertagia spp., together with some Haemonchus contortus, Trichostrongylus spp., and Dictyocaulus filaria. In 1 group, 5 sheep, which were treated twice with tetramisole at an interval of 4 weeks to remove gastro-intestinal nematodes and lungworms, showed an increase of 41% in weight gain and 3.2% in wool growth compared with an untreated group. Another group, which was treated with both tetramisole and rafoxanide to remove nematode and fluke infections, had a 143% increase in weight gain and 81% in wool growth compared with the control.

Animals

WNK1-OSR1 Signaling Regulates Angiogenesis-Mediated Metastasis towards Developing a Combinatorial Anti-Cancer Strategy.

Lysine-deficient protein kinase-1 (WNK1) is critical for both embryonic angiogenesis and tumor-induced angiogenesis. However, the downstream effectors of WNK1 during these processes remain ambiguous. In this study, we identified that oxidative stress responsive 1b (osr1b) is upregulated in endothelial cells in both embryonic and tumor-induced angiogenesis in zebrafish, accompanied by downregulation of protein phosphatase 2A (pp2a) subunit ppp2r1bb. In addition, wnk1a and osr1b are upregulated in two liver cancer transgenic fish models: [tert x p53-/-] and [HBx,src,p53-/-,RPIA], while ppp2r1bb is downregulated in [tert x p53-/-]. Furthermore, using HUVEC endothelial cells co-cultured with HepG2 hepatoma cells, we confirmed that WNK1 plays a critical role in the induction of hepatoma cell migration in both endothelial cells and hepatoma cells. Moreover, overexpression of OSR1 can rescue the reduced cell migration caused by shWNK1 knockdown in HUVEC cells, indicating OSR1 is downstream of WNK1 in endothelial cells promoting hepatoma cell migration. Overexpression of PPP2R1A can rescue the increased cell migration caused by WNK1 overexpression in HepG2, indicating that PPP2R1A is a downstream effector in hepatoma. The combinatorial treatment with WNK1 inhibitor (WNK463) and OSR1 inhibitor (Rafoxanide) plus oligo-fucoidan via oral gavage to feed [HBx,src,p53-/-,RPIA] transgenic fish exhibits much more significant anticancer efficacy than Regorafenib for advanced HCC. Importantly, oligo-fucoidan can reduce the cell senescence marker-IL-1β expression. Furthermore, oligo-fucoidan reduces the increased cell senescence-associated β-galactosidase activity in tert transgenic fish treated with WNK1-OSR1 inhibitors. Our results reveal the WNK1-OSR1-PPP2R1A axis plays a critical role in both endothelial and hepatoma cells during tumor-induced angiogenesis promoting cancer cell migration. By in vitro and in vivo experiments, we further uncover the molecular mechanisms of WNK1 and its downstream effectors during tumor-induced angiogenesis. Targeting WNK1-OSR1-mediated anti-angiogenesis and anti-cancer activity, the undesired inflammation response caused by inhibiting WNK1-OSR1 can be attenuated by the combination therapy with oligo-fucoidan and may improve the efficacy.

Animals

[The effect of newer anthelmintics on Fasciola hepatica in experimentally infected rats].

The reports deals with the results of testing seven new antihelminthics for Fasciola hepatica in the experimentally invaded Wistar rat. The greatest influence on juvenile flukes (2 and 4 weeks of age) was exerted by diamphenetid (Coriban) applied in a single dose of 100 mg kg-1. Hexachlorophene applied in the dose of 50 mg kg-1 showed the highest effect on sexually mature flukes. All the tested antihelminthics of the halogenated salicylanilide group were ineffective on juvenile stages and only slightly effective on mature F. hepatica flukes. It follows from the results that the effectiveness of some antifasciolics on laboratory animals need not always be in correlation with their effect in ruminants - hence it is necessary to verify the results obtained in laboratory animals and to check them on natural F. hepatica hosts.

Animals

Use of immunologic techniques to detect chemotherapeutic success in infections with Fasciola hepatica. II. The enzyme linked immunosorbent assay in infected rats and rabbits.

An enzyme-linked immunosorbent assay (ELISA) was developed using microtiter plates for the immunodiagnosis of fascioliasis in rats and rabbits using extracts of adult worms partially purified by gel filtration chromatography using Sephacryl S-200. Partial purification was necessary to eliminate cross-reactivity with antisera having antibodies to schistosomes. Soft polyvinyl plates clearly gave superior results over hard polystyrine plates. Titers rose by 4 weeks of infection in rats with fascioliasis, by 6 weeks in the case of rabbits, and remained high through at least 12 and 28 weeks, respectively. Titers drop rapidly when animals are successfully treated with a fasciolicidal drug at 4--6 weeks of infection. The results show that the ELISA can be employed for the serodiagnosis of fascioliasis in rats and rabbits and is useful for the prediction of chemotherapeutic success.

Animals