Browsing by Author "Elandalloussi, Laurence M."
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- Effect of antiprotozoal drugs on the proliferation of the bivalve parasite Perkinsus olseniPublication . Elandalloussi, Laurence M.; Leite, Ricardo; Rodrigues, Pedro; Afonso, Ricardo; Nunes, Patrícia A.; Cancela, LeonorThe protozoan parasite Perkinsus olseni causes severe losses among Ruditapes decussatus clams. The development of an in vitro culture of this parasite together with the use of a proliferation assay has provided the opportunity to screen for drug sensitivity of this parasite. Xenobiotics known for their antimalarial and antiprotozoal properties were tested against P. olseni. Only four of these drugs, namely cycloheximide, pyrimethamine, deferoxamine (DFO) and 2,2-bipyridyl (BIP), showed in vitro inhibitory effect on the parasite proliferation. Two in vivo experiments were designed to determine the effect of iron chelators on reducing P. olseni infection in clams. For this purpose, naturally infected clams from the Ria Formosa, Portugal, were exposed to DFO and BIP at various concentrations. In the first experiment, hemolymph samples were taken from each clam before and after treatment to determine the infection intensity and in the second experiment, clams were randomly distributed in groups of five and the parasite burden in treated and untreated groups was determined at the end of the experiment on the whole clam wet tissues. Only DFO was found to be effective in reducing in vivo P. olseni infections. In addition, acute toxicity of DFO and BIP has been determined and no mortality of Perkinsus-free clams was observed.
- Effect of the herbicide Roundup® on Perkinsus olseni in vitro proliferation and in vivo survival when infecting a permissive host, the Clam Ruditapes decussatusPublication . Elandalloussi, Laurence M.; Leite, Ricardo; Rodrigues, Pedro; Afonso, Ricardo; Cancela, LeonorCoastal habitats are increasingly being exposed to herbicide contamination from urban and agricultural catchments. Data on its toxicity on aquatic ecosystems, especially those based on sediment, are relatively scarce. This study aimed at investigating whether the susceptibility of an aquatic filter-feeding organism, the carpet-shell clam (Ruditapes decussatus) to the parasite Perkinsus olseni was influenced by the herbicide Roundup and its active ingredient glyphosate. The effect of Roundup and glyphosate on P. olseni in vitro proliferation was also evaluated and appeared to confirm the higher toxicity of Roundup when compared with technical grade glyphosate.
- Perkinsus Atlanticus – desenvolvimento de um método de diagnóstico para detecção de infecção em moluscos bivalvesPublication . Leite, Ricardo; Rodrigues, Pedro; Elandalloussi, Laurence M.; Afonso, Ricardo; Nunes, Patrícia A.; Cancela, LeonorDiferentes espécies do género Perkinsus, um protozoário parasita pertencente ao novo phylum Perkinsea, podem ser encontradas mundialmente e constituem uma séria ameaça aos moluscos bivalves como as ostras, amêijoas, abalones e vieiras, os quais têm um papel importante não só do ponto de vista comercial, mas também ecológico.
- Shikimate and folate pathways in the protozoan parasite, Perkinsus olseniPublication . Elandalloussi, Laurence M.; Rodrigues, Pedro; Afonso, Ricardo; Leite, Ricardo; Nunes, Patrícia A.; Cancela, LeonorWe have exploited the experimental accessibility of the protozoan parasite Perkinsus olseni and its similarities to apicomplexan parasites to investigate the influence of specific drugs on its proliferation. For this purpose, shikimate and folate pathways present an attractive target for parasitic therapy given their major differences with mammalian pathways. Glyphosate, a potent inhibitor of the shikimate pathway enzyme EPSP synthase inhibited the in vitro proliferation of P. olseni in a dose-dependent manner and this effectwas reversed by addition of chorismate, indicating the presence of a shikimate pathway. However, this effect was not antagonised by p-aminobenzoate or folic acid. Furthermore, antagonism was observed, via pyrimethamine to glyphosate inhibitory effect, suggesting that the shikimate pathway is not essential for the biosynthesis of folate precursors and is therefore crucial for another pathway downstream from chorismate. In addition, sulfadiazine, a well known inhibitor of dihydropteorate synthase, an enzyme of the folate biosynthetic pathway,had no inhibitory effect on P. olseni proliferation. In view of these results, the parasite does not appear to require the folate biosynthesis pathway for its survival and is most likely able to use exogenous folate. Even though pyrimethamine was found to inhibit P. atlanticus growth, this inhibitory effect could not be reversed by co-addition of folic acid. Therefore, we propose that the effect of pyrimethamine observed in this study results from the inhibition of a target other than dihydrofolate reductase. Similarly, proguanil target is likely to be separate from DHFR since only its metabolite cycloguanil has been shown to have inhibitory properties on DHFR.