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The present study aimed to determine the bioactivities of essential oils extracted from the leaves of <i>Paramignya trimera</i> and <i>Limnocitrus littoralis</i>, including cytotoxicity, antiviral, antibacterial, antimycotic, and antitrichomonas effects. Herein, it was indicated that <i>P. trimera</i> and <i>L. littoralis</i> oils showed no cytotoxicity on normal cells, namely MT-4, BHK-21, MDBK, and Vero-76. <i>P. trimera</i> oil (i) exhibited the strongest inhibition against <i>Staphylococcus aureus</i> with MIC and MLC values of 2% (<i>v/v</i>); (ii) showed MIC and MLC values of 8% (<i>v/v</i>) in <i>Candida parapsilosis</i>; and (iii) in the remaining strains, showed MIC and MLC values greater than or equal to 16% (<i>v/v</i>). On the other hand, <i>L. littoralis</i> oil (i) displayed the strongest inhibition against <i>Candida tropicalis</i> and <i>Candida parapsilosis</i> with 2% (<i>v/v</i>) of MIC and MLC; and (ii) in the remaining strains, possessed MIC and MLC greater than or equal to 16% (<i>v/v</i>). In addition, antitrichomonas activities of the oils were undertaken, showing IC₅₀, IC₉₀, MLC values, respectively, at 0.016%, 0.03%, and 0.06% (<i>v/v</i>) from <i>P. trimera,</i> and 0.03%, 0.06%, 0.12% (<i>v/v</i>) from <i>L. littoralis</i>, after 48 h of incubation. The oils were completely ineffective against ssRNA+ (HIV-1, YFV, BVDV, Sb-1, CV-B4), ssRNA- (RSV, VSV), dsRNA (Reo-1), and dsDNA (HSV-1, VV) viruses. This is the first report describing the cytotoxicity, antiviral, antibacterial, antimycotic, and antitrichomonas activities of the essential oils of <i>P. trimera</i> and <i>L. littoralis</i>.