Citrus essential oils and four enantiomeric pinenes against Culex pipiens (Diptera: Culicidae)

The aim of this study was to evaluate the toxicity of pinenes (entantiomers of α- and β-) and essential oils from Greek plants of the Rutaceae family against the mosquito larvae of Culex pipiens (Diptera: Culicidae). Essential oils were isolated by hydrodistillation from fruit peel of orange (Citrus sinensis L.), lemon (Citrus limon L.), and bitter orange (Citrus aurantium L.). The chemical composition was determined by gas chromatography/mass spectrometry (GC/MS) analysis. Citrus essential oils contained in high proportion limonene and in lower quantities p-menthane molecules and pinenes. The insecticidal action of these essential oils and entantiomers of their pinenes on mosquito larvae was evaluated. Plant essential oils exhibited strong toxicity against larvae with the LC₅₀ values ranging from 30.1 (lemon) to 51.5 mg/L (orange) depending on Citrus species and their composition. Finally, the LC₅₀ value of pinenes ranging from 36.53 to 66.52 mg/L indicated an enantioselective toxicity only for the β-pinene entantiomer.     

Acaricidal effect and chemical composition of essential oils extracted from Cuminum cyminum, Pimenta dioica and Ocimum basilicum against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

Acaricidal activity of essential oils extracted from cumin seeds (Cuminum cyminum), allspice berries (Pimenta dioica) and basil leaves (Ocimum basilicum) were tested on 10-day-old Rhipicephalus (Boophilus) microplus tick larvae using the LPT. Two-fold dilutions of the three essential oils were tested from a starting dilution of 20% down to 1.25%. Results showed a high toxicological effect for cumin, producing 100% mortality in all tested concentrations on R. microplus larvae. Similarly, allspice essential oil produced 100% mortality at all concentrations with the exception of a dramatic decrease at 1.25% concentration. Conversely, basil essential oil was not shown to be toxic against R. microplus larvae. The most common compounds detected by gas chromatography-mass spectrometry were as follows: cumin: cuminaldehyde (22.03%), γ-terpinene (15.69%) and 2-caren-10-al (12.89%); allspice: methyl eugenol (62.7%) and eugenol (8.3%); basil: linalool (30.61%) and estragole (20.04%). Results clearly indicate that C. cyminum and P. dioica essential oils can be used as an effective alternative for R. microplus tick control, and there is a high probability they can be used for other ticks affecting cattle in Mexico and throughout the world, thereby reducing the necessity for traditional and unfriendly synthetic acaricides.     

Chemical composition and larvicidal evaluation of Mentha, Salvia, and Melissa essential oils against the West Nile virus mosquito Culex pipiens

The volatile metabolites of wild-growing Mentha spicata, M. longifolia, M. suaveolens, Melissa officinalis, Salvia fruticosa, S. pomifera subsp. calycina, and S. pomifera subsp. pomifera from Greece were determined by gas chromatography and gas chromatography–mass spectrometry. The insecticidal properties of the analyzed essential oils were screened on Culex pipiens larvae. Additionally two of the main components of the essential oils, piperitenone oxide and 1,8-cineole were assayed against C. pipiens in order to define the affiliation between them and the larvicidal properties of the oils. The most effective oils were M. suaveolens (major constituent piperitenone oxide, 62.4%), M. spicata (piperitenone oxide, 35.7% and 1,8-cineole, 14.5%) and M. longifolia—Central Greece (piperitenone oxide, 33.4%; 1,8-cineole, 24.5% and trans-piperitone epoxide, 17.4%), which exhibited LC₅₀ values ranging from 47.88 to 59.33 mg l⁻¹. Medium activity revealed the oils of M. officinalis (terpin-4-ol, 15.8%; caryophyllene oxide, 13.2%; sabinene, 12.9%; β-pinene, 12.1%; and trans-caryophyllene, 10.2%), M. longifolia—Southern Greece (carvone, 54.7% and limonene 20.0%), S. pomifera subsp. pomifera (trans-caryophyllene, 22.5% and trans-thujone, 21.0%), S. pomifera subsp. calycina—West Southern Greece (trans-thujone, 56.1% and 1,8-cineole, 10.4%), and S. fruticosa—population 2 (camphor, 23.1%; α-pinene, 12.7%; and borneol, 12.6%), with LC₅₀ values ranging from 78.28 to 91.45 mg l⁻¹. S. pomifera subsp. calycina (Central Greece) essential oil (trans-thujone, 26.5% and cis-thujone, 12.0%) presented rather low activity (LC₅₀ values 140.42 mg l⁻¹), while S. fruticosa—population 1 (1,8-cineole, 31.4% and camphor, 22.6%) was the only inactive oil. Additionally, the constituent piperitenone oxide was found to be highly active (LC₅₀ values 9.95 mg l⁻¹), whereas 1,8-cineole revealed no toxicity.     

Effects of essential oils from Cymbopogon citratus (DC) Stapf., Lippia sidoides Cham., and Ocimum gratissimum L. on growth and ultrastructure of Leishmania chagasi promastigotes

The current therapy for leishmaniasis, which affects annually about 2 million people, is far from satisfactory. All available drugs require parenteral administration and are potentially toxic. Plant essential oils have been traditionally used in folk medicine and appear as valuable alternative source for chemotherapeutic compounds. In this study, we demonstrated the effect of essential oils from Cymbopogon citratus, Lippia sidoides, and Ocimum gratissimum on growth and ultrastructure of Leishmania chagasi promastigote forms. Steam distillation was used to isolate the essential oils, and their constituents were characterized by gas chromatography coupled to mass spectrometry and nuclear magnetic resonance. All essential oils showed in vitro inhibitory action on L. chagasi promastigotes growth in a dose-dependent way, with IC₅₀/72 h of 45, 89, and 75 μg/mL for C. citratus, L. sidoides, and O. gratissimum, respectively. Drastic morphological alterations were observed in all essential oil-treated parasites, including cell swelling, accumulation of lipid droplets in the cytoplasm, and increase of acidocalcisome volume. Furthermore, aberrant-shaped cells with multi-septate body were observed by scanning electron microscopy, suggesting an additional effect on cytokinesis. Taken together, our data show that these essential oils affect the parasite viability being the C. citratus essential oil the most effective against L. chagasi.     

Essential oil composition and larvicidal activity of six Mediterranean aromatic plants against the mosquito Aedes albopictus (Diptera: Culicidae)

Laboratory bioassays on insecticidal activity of essential oils (EOs) extracted from six Mediterranean plants (Achillea millefolium, Lavandula angustifolia, Helichrysum italicum, Foeniculum vulgare, Myrtus communis, and Rosmarinus officinalis) were carried out against the larvae of the Culicidae mosquito Aedes albopictus. The chemical composition of the six EOs was also investigated. Results from applications showed that all tested oils had insecticidal activity, with differences in mortality rates as a function of both oil and dosage. At the highest dosage (300 ppm), EOs from H. italicum, A. millefolium, and F. vulgare caused higher mortality than the other three oils, with mortality rates ranging from 98.3% to 100%. M. communis EO induced only 36.7% larval mortality at the highest dosage (300 ppm), a similar value to those recorded at the same dosage by using R. officinalis and L. angustifolia (51.7% and 55%, respectively). Identified compounds ranged from 91% to 99%. The analyzed EOs had higher content of monoterpenoids (80–99%) than sesquiterpenes (1–15%), and they can be categorized into three groups on the basis of their composition. Few EOs showed the hydrocarbon sesquiterpenes, and these volatile compounds were generally predominant in comparison with the oxygenated forms, which were detected in lower quantities only in H. italicum (1.80%) and in M. communis (1%).     

Insecticidal Evaluation of essential oils of <Emphasis Type="BoldItalic">Citrus sinensis</Emphasis> L. (Myrtales: Myrtaceae) against housefly, <Emphasis Type="BoldItalic">Musca domestica</Emphasis> L. (Diptera: Muscidae)

The housefly, Musca domestica L., is one of the most common insects, associated with vectoring of various etiological agents. In order to search for effective control agent, the essential oil of sweet orange [Citrus sinensis (L.) Osbeck] was evaluated for its insecticidal activity against the larvae and pupae of housefly using contact toxicity and fumigation bioassays. In the contact toxicity assay, lethal concentration, LC₅₀ of C. sinensis essential oil against housefly larvae, varied between 3.93 and 0.71 μl/cm² for different observation days, while lethal time, LT₅₀, varied between 5.8 to 2.3 days. Mortality of larvae were significant with different concentrations (F = 2.79, df = 4, P < 0.05) and time (F = 6.69, df = 3, P < 0.01). In fumigant assay for housefly larvae, LC₅₀ of 71.2 and 52.6 μl/l was obtained in 24 and 48 h, respectively. Scanning electron microscopy of oil treated larvae revealed extreme dehydration and surface distortion while control larvae were free from any of the above symptoms and presented smooth surface, conforming effect of essential oil on housefly larvae. Percentage inhibition rate of oil against housefly pupae was 27.3–72.7% for contact toxicity and 46.4–100% for fumigation assay. Compositional analysis of C. sinensis essential oil using gas chromatography/mass spectrometry (GC-MS) revealed d-limonene (73.24%), α-pinene (5.86%) and myrcene (4.45%) as major components whereas its vapour profile (solid-phase micro extraction-GC/MS) was dominated by d-limonene at 92.57%. Significant activity of C. sinensis essential oil against larvae and pupae of housefly, pave the way for its use as eco-friendly housefly control measure.     

Acaricidal activity of the essential oils from Eucalyptus citriodora and Cymbopogon nardus on larvae of Amblyomma cajennense (Acari: Ixodidae) and Anocentor nitens (Acari: Ixodidae)

The present study evaluated the acaricidal activity of essential oils from Eucalyptus citriodora and Cymbopogon nardus on non-engorged larvae of Amblyomma cajennense and Anocentor nitens. In order to carry out the study, six groups were formed, each concentration being a treatment (6.25%, 12.5%, 25%, and 50%, respectively) and also with the creation of a control group (distilled water) and a positive control (Deltametrine). For each treatment, approximately 100 larvae of these ticks were placed onto filter papers (2 × 2 cm) impregnated with the concentrations used to test. Next, the envelopes were closed bearing inside the filter paper with measurements of 6 × 6 cm. For each group, six repetitions were performed, and after 24 h live and dead larvae were counted. This procedure was carried out for two essential oils on the two species of ticks. For A. cajennense, the acaricide efficacy of E. citriodora oil was of 10.8%, 35.3%, 34.5%, and 53.1%, whereas the efficacy of C. nardus was of 0.0%, 0.0%, 0.0%, and 61.1% at concentrations of 6.25%, 12.5%, 25.0%, and 50.0%, respectively. In relation to A. nitens, the acaricide efficacy of E. citriodora oil was of 20.1%, 84.5%, 89.2%, and 100.0%, whereas the efficacy of C. nardus was of 0.0%, 90.8%, 100.0%, and 100.0% at concentrations of 6.25%, 12.5%, 25.0%, and 50.0%, respectively. The results indicate that the essential oils tested showed a promising acaricidal activity mainly on A. nitens larvae.     

Evaluation of bioefficacy of three Citrus essential oils against the dengue vector Aedes albopictus (Diptera: Culicidae) in correlation to their components enantiomeric distribution

Laboratory experiments were conducted to study the bioefficacy against Ae. albopictus of three Citrus essential oils, derived from peels of Citrus sinensis, Citrus limon, and Citrus paradise and of their components. Chiral gas chromatography analysis revealed the dominant occurrence of R-(+)-limonene and (?)-??-pinene in all three essential oils while in the case of lemon oil ??-terpinene, neral, and geranial detected also among other components. The tested Citrus essential oils were toxic against mosquito larvae with LC₅₀ values ranging from 25.03 to 37.03?mg?l^?1. Among citrus essential oils components tested, ??-terpinene was the most toxic (LC₅₀?=?20.21?mg?l^?1) followed by both enantiomeric forms of limonene (LC₅₀?=?35.99 and 34.89?mg?l^?1, for R-(+)-limonene and S-(?)-limonene, respectively). The delayed toxic effects after exposure of larvae to sublethal (LC₅₀) doses were also investigated for citrus essential oils and their major component R-(+)-limonene, indicating a significant reduction of pupal survival. In repellent bioassays, lemon essential oil, S-(?)-limonene, citral (mixture of neral\geranial) and (+)-??-pinene were the most effective compared with other citrus essential oils and components against adult mosquitoes. Repellent bioassays also revealed that limonenes and ??-pinenes showed an isomer dependence repellent activity. Finally, according to enantiomeric distribution of limonene and ??- and ??-pinene, the repellency of lemon essential oil is possibly attributed to the presence of citral.     

Acaricidal activity and chemical composition of the essential oil from three Piper species

The chemical composition of the essential oil obtained by hydrodistillation of the aerial parts of Piper amalago, Piper mikanianum, and Piper xylosteoides was elucidated by gas chromatography (GC) and GC/mass spectrometry analyses. P. mikanianum and P. xylosteoides essential oils presented phenylpropanoids as their main compounds (67.89% and 48.53%, respectively) whereas P. amalago was rich in monoterpene and sesquiterpene hydrocarbons (84.95%). The essential oils obtained were investigated for their effect on newly hatched larvae of the cattle tick, Rhipicephalus (Boophilus) microplus. The essential oil of P. mikanianum (LC₅₀ 2.33 μL/mL) was more active than that of P. xylosteoides (LC₅₀ 6.15 μL/mL) against the larvae, while the oil of P. amalago was inactive. These results suggest that phenylpropanoids, mainly apiol and safrole, are responsible for the acaricidal activity.     

Antiplasmodial volatile extracts from Cleistopholis patens Engler & Diels and Uvariastrum pierreanum Engl. (Engl. & Diels) (Annonaceae) growing in Cameroon

In a search for alternative treatment for malaria, plant-derived essential oils extracted from the stem barks and leaves of Cleistopholis patens and Uvariastrum pierreanum (Annonaceae) were evaluated in vitro for antiplasmodial activity against the W2 strain of Plasmodium falciparum. The oils were obtained from 500 g each of stem barks and leaves, respectively, by hydrodistillation, using a Clevenger-type apparatus with the following yields: 0.23% and 0.19% for C. patens and 0.1% and 0.3% for U. pierreanum (w/w relative to dried material weight). Analysis of 10% (v/v) oil in hexane by gas chromatography and mass spectrometry identified only terpenoids in the oils, with over 81% sesquiterpene hydrocarbons in C. patens extracts and U. pierreanum stem bark oil, while the leaf oil from the latter species was found to contain a majority of monoterpenes. For C. patens, the major components were α-copaene, δ-cadinene, and germacrene D for the stem bark oil and β-caryophyllene, germacrene D, and germacrene B for the leaf oil. The stem bark oil of U. pierreanum was found to contain mainly β-bisabolene and α-bisabolol, while α- and β-pinenes were more abundant in the leaf extract. Concentrations of oils obtained by diluting 1-mg/mL stock solutions were tested against P. falciparum in culture. The oils were active, with IC₅₀ values of 9.19 and 15.19 μg/mL for the stem bark and leaf oils, respectively, of C. patens and 6.08 and 13.96 μg/mL, respectively, for those from U. pierreanum. These results indicate that essential oils may offer a promising alternative for the development of new antimalarials.     

Validation of models to estimate the fumigant and larvicidal activity of Eucalyptus essential oils against Aedes aegypti (Diptera: Culicidae)

The aim of this work is to validate the pre-existing models that relate the larvicidal and adulticidal activities of the Eucalyptus essential oils on Aedes aegypti. Previous works at our laboratory described that the larvicidal activity of Eucalyptus essential oils can be estimated from the relative concentration of two main components (p-cymene and 1,8-cineole) and that the adulticidal effectiveness can be explained, to a great extent, by the presence of large amounts of the component 1,8-cineole in it. In general, the results show that the higher adulticidal effect of essential oils the lower their larvicidal activity. Fresh leaves was harvested and distilled. Once the essential oil was obtained, the chemical composition was analysed, evaluating the biological activity of 15 species of the genus Eucalyptus (Eucalyptus badjensis Beuzev and Welch, Eucalyptus badjensis × nitens, Eucalyptus benthamii var Benthamii Maiden and Cambage, Eucalyptus benthamii var dorrigoensis Maiden and Cambage, Eucalyptus botryoides Smith, Eucalyptus dalrympleana Maiden, Eucalyptus fastigata Deane and Maiden, Eucalyptus nobilis L.A.S. Johnson and K.D.Hill, Eucalyptus polybractea R. Baker, Eucalyptus radiata ssp radiata Sieber ex Spreng, Eucalyptus resinifera Smith, Eucalyptus robertsonii Blakely, Eucalyptus robusta Smith, Eucalyptus rubida Deane and Maiden, Eucalyptus smithii R. Baker). Essential oils of these plant species were used for the validation of equations from preexistent models, in which observed and estimated values of the biological activity were compared. The regression analysis showed a strong validation of the models, re-stating the trends previously observed. The models were expressed as follows: A, fumigant activity [KT_50(min) = 10.65–0.076 × 1,8-cineole (%)](p < 0.01; F, 397; R ², 0.79); B, larval mortality (%)_(40 ppm) = 103.85 + 0.482 × p-cymene (%) − 0.363 × α-pinene (%) − 1.07 × 1,8-cineole (%) (p < 0.01; F, 300; R ², 0.90). These results confirmed the importance of the mayor components in the biological activity of Eucalyptus essential oils on A. aegypti. However, it is worth mentioning that two or three species differ in the data estimated by the models, and these biological activity results coincide with the presence of minor differential components in the essential oils. According to what was previously mentioned, it can be inferred that the model is able to estimate very closely the biological activity of essential oils of Eucalyptus on A. aegypti.     

Effects of the essential oils of Lippia turbinata and Lippia polystachya (Verbenaceae) on the temporal pattern of locomotion of the mosquito Culex quinquefasciatus (Diptera: Culicidae) larvae

The essential oils (EO) of Lippia turbinata (TUR) and Lippia polystachya (POL) have shown lethal effects against mosquito larvae. The present work evaluated whether these EO at doses ranging from sublethal to lethal (20, 40 and 80 ppm) modify the temporal pattern of locomotion of Culex quinquefasciatus larvae. Larvae were individually placed in glass boxes, and their activity recorded at 0.3 s intervals during 40 min. Individuals treated with doses >40 ppm of either EO significantly decreased their ambulation speed and the percentage of total time ambulating compared to controls. TUR 80 ppm decreased their ambulation even sooner than POL 80 ppm, when compared to their respective controls. These findings are consistent with the neurotoxic effect against insects attributed to α-Thujone, a main component of both EO. A detrended fluctuation fractal analysis evaluating the complexity and organisation of the temporal pattern of locomotion showed fractal patterns in all animals. Both sublethal and lethal doses of TUR and POL increased the complexity of ambulation. Interestingly, for POL 20 ppm, an increase in complexity was observed, while no changes in general activity were detected, suggesting that fractal analysis may be more sensitive to detect behavioural changes than general activity evaluation.     

Chemical composition and larvicidal activity of essential oil from <Emphasis Type="Italic">Mentha spicata</Emphasis> (Linn.) against three mosquito species

Mosquitoes are blood-feeding insects and serve as the most important vectors for spreading human diseases such as malaria, yellow fever, dengue fever, and filariasis. The continued use of synthetic insecticides has resulted in resistance in mosquitoes. Synthetic insecticides are toxic and affect the environment by contaminating soil, water, and air, and then natural products may be an alternative to synthetic insecticides because they are effective, biodegradable, eco-friendly, and safe to environment. Botanical origin may serve as suitable alternative biocontrol techniques in the future. Mentha spicata, an edible and medicinal plant, is chiefly distributed in Southeast Asia and South Asia. In the present study, the toxicity of mosquito larvicidal activity of leaf essential oil (EO) and their major chemical constituents from Mentha spicata against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The chemical composition of the leaf EO was analyzed using gas chromatography–mass spectroscopy (GC-MS). GC-MS revealed that the EO of M. spicata contained 18 compounds. The major chemical components identified were carvone (48.60%), cis-carveol (21.30%), and limonene (11.30%). The EO had a significant toxic effect against early third-stage larvae of C. quinquefasciatus, A. aegypti, and A. stephensi with LC₅₀ values of 62.62, 56.08, and 49.71 ppm and LC₉₀ values of 118.70, 110.28, and 100.99 ppm, respectively. The three major pure constituents extracted from the M. spicata leaf EO were also tested individually against three mosquito larvae. The LC₅₀ values of carvone, cis-carveol, and limonene appeared to be most effective against A. stephensi (LC₅₀ 19.33, 28.50, and 8.83 ppm) followed by A. aegypti (LC₅₀ 23.69, 32.88, and 12.01 ppm), and C. quinquefasciatus (LC₅₀ 25.47, 35.20, and 14.07 ppm). The results could be useful in search for newer, safer, and more effective natural larvicidal agents against C. quinquefasciatus, A. aegypti, and A. stephensi.     

Oviposition-altering and ovicidal potentials of five essential oils against female adults of the dengue vector, <Emphasis Type="BoldItalic">Aedes aegypti</Emphasis> L.

The oviposition deterrence and ovicidal potential of five different essential oils, peppermint oil (Mentha piperita), basil oil (Ocimum basilicum), rosemary oil (Rosemarinus officinalis), citronella oil (Cymbopogon nardus), and celery seed oil (Apium graveolens), were assessed against female adults of the dengue vector, Aedes aegypti L. Multiple concentration tests were carried out where cups containing 1 mL of different concentrations (100%, 10%, 1%, 0.1%) of the oils and 199 mL of water were used for oviposition. The number of eggs laid and the larvae hatched in each cup were scored to evaluate the oviposition deterrent and ovicidal potentials of the oils. Our investigations revealed that the addition of 100% oil (pure oil) caused complete oviposition deterrence except in A. graveolens which resulted in 75% effective repellency. The use of 10% oil resulted in the maximum deterrence of 97.5% as shown by the M. piperita oil while other oils caused 36–97% oviposition deterrence as against the control. The oviposition medium with 1% oil showed decreased deterrent potential with 30–64% effective repellency, the M. piperita oil being exceptional. However, as the concentrations of the oil were reduced further to 0.1%, the least effective oil observed was A. graveolens (25% ER). Also, the M. piperita oil showed much reduced activity (40%) as compared to the control, while the other oils exhibited 51–58% repellency to oviposition. The studies on the ovicidal effects of these oils revealed that the eggs laid in the water with 100% essential oils did not hatch at all, whereas when 10% oils were used, only the R. officinalis oil resulted in 28% egg hatch. At lower concentrations (1%), the oils of M. piperita, O. basilicum, and C. nardus showed complete egg mortality while those of A. graveolens and R. officinalis resulted in 71% and 34% egg hatches, respectively. When used at 0.1%, the O. basilicum oil was found to be the only effective oil with 100% egg mortality, whereas other oils resulted in 16–76% egg mortality, the least mortality caused by the A. graveolens oil. These results suggest that these essential oils can be employed in a resistance-management program against A. aegypti. Further detailed research is needed to identify the active ingredient in the extracts and implement the effective mosquito management program.     

Life cycle of Ixodes luciae (Acari: Ixodidae) in the laboratory

The life cycle of Ixodes luciae was evaluated for five consecutive generations in the laboratory. Wild mice Calomys callosus and laboratory rats Rattus norvegicus were used as hosts for larvae and nymphs. For adult ticks, opossums Didelphis aurita were used as hosts. Off-host developmental periods were observed in an incubator at 27°C and 95% RH. The life cycle of I. luciae lasted 95–97 days, excluding prefeeding periods. C. callosus, one of the natural host species for I. luciae immature stages, was shown to be much more suitable than the artificial host R. norvegicus. Significantly (P < 0.05), more larvae and nymphs successfully fed on C. callosus than on R. norvegicus. When tick-na?ve C. callosus were exposed to three consecutive larval infestations at 24-day intervals, recovery of engorged larvae were greater in the second and third infestations, indicating that previous infestations did not induce acquired resistance to ticks. Larval feeding period typically varied from 5 to 10 days on R. norvegicus, but was significantly (P < 0.05), longer on C. callosus (range, 7–34 days). The majority (71.7%) of I. luciae adult females successfully fed and oviposited after exposed to D. aurita. Mean engorged weight (581.9 mg; range, 237.1–796.0 mg) of these females were much higher than those previously reported for other New World Ixodes species. Our results are in accordance to the current literature that appoints opossums Didelphidae and small rodents (e.g., C. callosus) natural hosts for I. luciae immature and adult stages, respectively.     

Antiparasitic activity of two Lavandula essential oils against Giardia duodenalis, Trichomonas vaginalis and Hexamita inflata

Two essential oils derived from Lavandula angustifolia and Lavandula × intermedia were investigated for any antiparasitic activity against the human protozoal pathogens Giardia duodenalis and Trichomonas vaginalis and the fish pathogen Hexamita inflata: all of which have significant infection and economic impacts. The study has demonstrated that low (≤1%) concentrations of L. angustifolia and L. × intermedia oil can completely eliminate T. vaginalis, G. duodenalis and H. inflata in vitro. At 0.1% concentration, L. angustifolia oil was found to be slightly more effective than L × intermedia oil against G. duodenalis and H. inflata. The potential applications are discussed.     

Evaluation of time toxicity, residual effect, and growth-inhibiting property of Carapa guianensis and Copaifera sp. in Aedes aegypti

Oils of Carapa guianensis and Copaifera spp. are well-known in the Amazonian region as natural insect repellents, and studies have reported their efficiency as larvicide against some mosquito species. However, toxicity persistence and effect on mosquito development have not yet been evaluated. Thus, the objective of this study was to evaluate the initial time of larvicidal activity, residual effect, and the effect of very low concentrations of these oils on Aedes aegypti. Different concentrations of the oils were used to evaluate the initial time of larval mortality and residual effect, as well as, the development of larvae, pupae, and adults. Results demonstrated that the lethal effect started mainly between the first 2 and 3 h of larvae exposure to oils, when using concentrations which ranged from 500 mg/L of C. guianensis and 90 mg/L of Copaifera sp. The toxic effect remained with total efficiency (100% mortality) until the sixth day for Copaifera sp. and 12th day for C. guianensis. When using sublethal dosages (ranging from 140 mg/L of C. guianensis to 26 mg/L of Copaifera sp.) mortality was observed after the larval molt. Also, imperfection of pupae and adult development and unsuccessful emergence of adults were observed. A product of botanical origin that could break the development of immature stage of mosquitoes and inhibit the emergence of adults should be essential in vector control. Thus, our results provide new information for a better understanding in using C. guianensis and Copaifera sp. oils with a potential to be used as a natural insecticide.     

Oviposition deterrent and ovicidal activities of seven herbal essential oils against female adults of housefly,Musca domestica L.

The oviposition deterrent and ovicidal of seven herbal essential oils derived from Citrus sinensis, Cymbopogon citratus, Eucalyptus glubulus, Illicium verum, Lavandula angustifolia, Mentha piperita, and Zingiber cussumunar were assessed against the gravid female of housefly, Musca domestica L., under laboratory conditions and compared with commercial insecticide (10 % w/v cypermethrin). They were assayed at three concentrations (1.0, 5.0, and 10.0 %) where plastic cups containing 1 ml of desired oil concentration and cotton pad soaked with 10 ml of milk solution (10 % w/v) were used as oviposition substrate. The 0.1 ml of deferent concentrations was dropped on ten housefly eggs, which were used for ovicidal activity. The number of eggs laid and the hatched larvae in each cup was recorded to evaluate the oviposition deterrent and ovicidal activities of the herbal essential oils. High concentration (10 %) of herbal essential oils showed high percent effective repellency (ER). The 10 % I. verum oil caused complete oviposition deterrence (100 % ER, oviposition activity index (OAI)?=??1.0), followed by Z. cussumunar, M. piperita, L. angustifolia, C. citratus, C. sinensis, and E. glubulus oils with 97.20, 88.55, 88.14, 87.93, 76.68, and 57.00 % ER, respectively. As the concentration of herbal essential oils increased from 1.0, 5.0, and up to 10.0 % concentration, the hatching rate decreased. Ten percent I. verum oil gave the maximum inhibiting rate at 97.3 % (LC₅₀ value of 6.85 %); in addition, the other herbal essential oils showed the minimum inhibiting rate of 3.3–22.7 %. On the other hand, cypermethrin 10 % w/v showed complete oviposition deterrence (100 % ER, OAI?=??1.0) and ovicidal activity (100 % inhibiting rate). Our data showed that I. verum oil have high potential of oviposition deterrence and ovicide housefly control.     

Effect of oregano (<Emphasis Type="Italic">Origanum vulgare</Emphasis> L.) and thyme (<Emphasis Type="Italic">Thymus vulgaris</Emphasis> L.) essential oils on <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis> (Protozoa: Kinetoplastida) growth and ultrastructure

In the present work, we have investigated the effect of essential oils obtained from Origanum vulgare L. (oregano) and Thymus vulgaris L. (thyme) on growth and ultrastructure of diverse evolutive forms of Trypanosoma cruzi. Culture epimastigotes and bloodstream trypomastigotes were incubated for 24 h with different concentrations of oregano or thyme essential oils and with thymol (the main constituent of thyme), and the inhibitory concentration (IC)₅₀ was determined by cell counting. Crude extract of oregano essential oil inhibited epimastigote growth (IC₅₀/24 h = 175 μg/ml) and also induced trypomastigote lysis (IC₅₀/24 h = 115 μg/ml). Thyme essential oil presented IC₅₀/24 h values of 77 μg/ml for epimastigotes and 38 μg/ml for trypomastigotes, while treatment with thymol resulted in an IC₅₀/24 h of 62 μg/ml for epimastigotes and 53 μg/ml for trypomastigotes. Scanning electron microscopy of treated cells showed few morphological alterations at the plasma membrane. Observation by transmission electron microscopy showed cytoplasmic swelling with occasional morphological alterations in plasma and flagellar membrane. Our data indicate that oregano and thyme essential oils are effective against T. cruzi, with higher activity of thyme, and that thymol may be the main component responsible for the trypanocidal activity.     

Eucalyptus essential oil toxicity against permethrin-resistant Pediculus humanus capitis (Phthiraptera: Pediculidae)

During the past decades, chemical control against the head louse Pediculus humanus capitis De Geer has been based in the application of products containing permethrin. The repetitive overuse of pediculicides has resulted in the development of high levels of resistance to one or more of these products worldwide. Essential oils obtained from aromatic plants like Eucalyptus are good and safe alternatives due to their low toxicity to mammals and easy biodegradability. In the present study, we reported the chemical composition of Eucalyptus dunnii and Eucalyptus gunni, and the fumigant activity of five Eucalyptus essential oils and their main compounds against permethrin-resistant head lice from Argentina. The most effective essential oils were Eucalyptus sideroxylon, Eucalyptus globulus ssp globulus, and Eucalyptus globulus ssp maidenii, with knockdown time 50% (KT₅₀) values of 24.75, 27.73, and 31.39 min. A linear regression analysis between percentage of 1,8-Cineole and KT₅₀ values of the essential oils showed a significant correlation at a p < 0.01. Since Eucalyptus essential oils showed to be effective against head lice and are classified as safer compounds, they can be employed into pediculicide formulations.