Infectious diseases are the major health problems in the world particularly in developingrncountries. To combat this problem various drugs, antibiotics and vaccines have been developedrnthrough time, but these successes have been challenged by the emergence of drug resistantrnpathogens. In Ethiopia, most people use traditional medicinal plants which are the natural sourcernof medicine to treat different diseases. However, traditional healers in this country applyrnmedicinal plants to heal different infections without scientific prescription and optimization ofrnthe right doses. This study was conducted to investigate antiplasmodial, antibacterial, antifungalrnproperties, evaluate acute toxicity and possibly isolate the active compounds of two widely usedrnmedicinal plants: Rumex nepalensis Spreng and Centella asiatica L. The two plants wererncollected from Dejen District, Northen part of Ethiopia, and air dried in shade. The specific plantrnparts (leaves, roots. aerial parts) were separately ground into powder. Each plant powder wasrnsoaked in water, methanol and hexane in separate Erlenmeyer flasks and placed on shaker of 120rnrpm for 72 hours. The extracts were concentrated in rotary evaporator and by lyophilizerrn(aqueous extract). The crude extracts of water, methanol and hexane of the two plants wererntested for antiplasmodial, antimicrobial activities, acute toxicity and screened forrnphytochemicals. The higher antimicrobial performing extract, methanol extracts of the roots ofrnR. nepalensis and further fractionnated by solvent-solvent extraction (hexane, chloroform,rnethanol, water and methanol) and these fractions were evaluated on pathogenic microorganismsrn(Plasmodium berghei, Esherchia coli, Pseudomonas aeruginosa, Shigella flexneri andrnSalmonella typhimurium, Klebsiella pneumonia, Candida albicans). Similarly the best fraction,rnethanol fraction, was further fractionnated by column chromatography (CC) and tested forrnantiplasmodial and antimicrobial activities. Furthermore, column Chromatographic profiles of the roots of R. nepalensis were analyzed by thin layer chromatography (TLC). The crudernmethanol and hexane extracts of the root, the leaf extracts of R. nepalensis and the aerial parts ofrnC. asiatica at the highest concentration of 5000 mg/kg BW was not toxic to the albino mice.rnFurthermore the phytochemical screening of the R. nepalensis revealed the presence ofrnflavonoids, alkaloids, glycosides, triterpenoids, sterols, saponin and tannin. On the other hand,rnphytochemical screening of C. asiatica showed the presence of flavonoids, alkaloids, glycosidesrnand saponin, triterpenoids and sterols but tannins were not clearly observed. CC and TLCrnprofiles of ethanol fraction of the roots of R. nepalensis showed the presence of chrysophanol,rnemodin and 5 undefined compounds. The crude extracts of water, methanol and hexane of therntwo plants had shown suppression to Plasmodium berghei in vivo in mice. Particularly, the crudernextracts of water, methanol and hexane of the roots of R. nepalensis had significantly higher day-rn4 suppressive effect 52.00 %, 59.90 % and 39.30 % respectively than the negative control (2%rnDMSO). But all extracts had significantly lower day-4 suppressive level than the positiverncontrol, chloroquine in P. berghei infected albino mice. The ethanol fraction of the roots of R.rnnepalensis showed higher plasmodium percent suppression in vivo in albino mice (70.08 %)rncompared to water (54.31 %), chloroform (19.61 %) and methanol (10.27 %) suppression at 500rnmg/kg body weight (BW). Similarly the mean survival time value of the mice administered withrnethanol fraction was significantly higher than all extracts (water, chloroform and methanol) atrn500 mg/kg BW and negative control, 2% DMSO treated mice. More specificaly the ethanolrnfraction of the roots of R. nepalensis fractionnated by column chromatography in ethyl acetate:rnmethanol (1:1) v/v showed 73.66% plasmodium suppression in vivo in albino mice at 300 mg/kgrnBW. The re-fractionnated ethyl acetate: methanol fraction by column chromatography inrnmethanol increased its suppression to 79.66% at 150 mg/kg BW. In the antimicrobial test, the ethanol fraction of the roots of R. nepalensis showed zone of inhibition for P. aeruginosarn(19.33Â±0.33 mm), S. aureus (18.03Â±0.14 mm) and S. flexneri (12.77Â±0.15 mm) and S.rntyphimurium (8.33Â±0.33 mm) at 6 mg/well by using agar well diffusion method compared to thernnegative control (2% DMSO) which have no inhibition zone on the tested bacteria. However, thernpositive control (teteracyclin and chloramphenicol) significantly higher bacterial inhibition thanrnethanol fraction of the roots of R. nepalensis. From the result obtained it can be concluded thatrnthe five undefined compounds played a role in synergizing the atiplasmodial, antibacterial andrnantifungal activities of the roots of R. nepalensis. In conclusion this work has demonstrated thernhigh potential of R. nepalensis and C. asiatica as sources for further exploitation ofrnantiplasmodial, antibacterial and for use in herbal medicine.rnKey words: antibacterial, antifungal, Plasmodium suppression, phytochemical screening,rntoxicity