Acaricidal activities of some essential oils and their monoterpenoidal constituents against house dust mite, Dermatophagoidespteronyssinus (Acari: Pyroglyphidae)

Acaricidal activities of some essential oils and their monoterpenoidal constituents against house dust mite, Dermatophagoides pteronyssinus (Acari: Pyroglyphidae) 1 first-author †E-mail:saad_elzemity@yahoo.com †E-mail:saad_elzemity@yahoo.com saad_elzemity@yahoo.com 2 2 2 Department of Pesticide Chemistry, Faculty of Agriculture, Alexandria University, Alexandria 21-545, Egypt Department of Pesticide Chemistry, Faculty of Agriculture, Alexandria University, Alexandria 21-545, Egypt Department of Economic Entomology, Faculty of Agriculture, Alexandria University, Alexandria 21-545, Egypt Department of Economic Entomology, Faculty of Agriculture, Alexandria University, Alexandria 21-545, Egypt The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus . Five concentrations were used over two different time intervals 24 and 48 h under laboratory conditions. In general, it was noticed that the acaricidal effect based on LC 50 of either essential oils or monoterpenoids against the mite was time dependant. The LC 50 values were decreased by increasing of exposure time. Clove, matrecary, chenopodium, rosemary, eucalyptus and caraway oils were shown to have high activity. As for the monoterpenoids, cinnamaldehyde and chlorothymol were found to be the most effective followed by citronellol. This study suggests the use of the essential oils and their major constituents as ecofriendly biodegradable agents for the control of house dust mite, D . pteronyssinus . Natural acaricides Essential oils Monoterpenoids House dust mite Dermatophagoides pteronyssinus INTRODUCTION 1 Among several species of the family Pyroglyphidae, Dermatophagoides pteronyssinus and D . farinae , have been found to be the predominant mites of household dust accounting for about 80%~90% of the total mite populations and are important sources of allergens worldwide inside homes in humid geographic areas (Hallas, 1991 1991 ; Arlian et al., 1992 1992 ). Both house dust mites are found in homes in various areas of Egypt (Rezk, 2004 2004 ; Rezk et al., 1996 1996 ). They are a major cause of respiratory allergies including asthma (Colloff et al., 1992 1992 ; Solarz, 2001a 2001a ; 2001b 2001b ). They are also identified as etiologic agent of sensitization and asthma-triggering in children (Lau et al., 1989 1989 ; Arshad and Hide, 1992 1992 ). Control of house dust mite is usually done by three different approaches: the first is a physical method such as exceptional cleaning standards (Adilah et al., 1997 1997 ; Nishioka et al., 1998 1998 ), the second is reducing the humidity levels (Bischoff et al., 1998 1998 ) and the third reducing allergens by effective acaricides (Mitchell et al., 1985 1985 ; Green et al., 1989 1989 ). The latter option must only be employed after careful consideration. Although good control was obtained by the currently used synthetic acaricides (benzylbenzoate and tannic acid), the risk to human health would be a potential problem. This problem has led to research efforts to develop safer and efficient alternatives in controlling mites in the indoor environment. Thus, the use of plant-derived acaricides has drawn attention and has been considered as a promising alternative to chemical acaricides (Miyazaki et al., 1989 1989 ; McDonald and Tovey, 1993 1993 ; Tovey and McDonald, 1997 1997 ; Kim, 2001 2001 ; Kim et al., 2003 2003 ; Rezk and Gadelhak, 2004 2004 ; Rembold, 2005 2005 ). Comparative data obtained by McDonald and Tovey ( 1993 1993 ) showed that citronella oil and tee tree oil were as effective as 0.5% benzyl benzoate (synthetic acaricide). Recently, Lee ( 2004 2004 ) compared the oil of Foeniculum vulgare and five of its commercial constituents (monoterpenoids) with benzyl benzoate against D. farinae and D. pteronyssinus . He found that both fenchone isomers and anisaldehyde were more effective than benzyl benzoate. However, thymol was as effective as benzyl benzoate as acaricide. Therefore, the present work has been devoted to evaluating the miticidal activity of some commonly available essential oils and their major constituents against the house dust mite, D. pteronyssinus , the most important vector in asthma, in order to find new natural miticides. MATERIALS AND METHODS 1 Stock culture of house dust mite 2 The house dust mite, D . pteronyssinus , was isolated from mattress dust and reared on a finely-ground mixture of dust, dried yeast and dried milk (1:1:0.5) in complete darkness (de Saint Georges-Gridelet, 1987 1987 ). Stock jars were kept in an incubator at an average temperature of (25±2) °C and relative humidity of (80±5)%. Considerable numbers of different stages were available after five months for experimentation. Plant essential oils 2 Fourteen essential oils were used in the present study namely: caraway, chenopodium, cinnamon, clove, eucalyptus, rosemary, garlic, geranium, lemon grass, matrecary, peppermint, rose, fennel and thyme. These oils were supplied by the Faculty of Pharmacy, University of Alexandria, Egypt. Identification and determination of essential oils composition 2 The essential oil constituents (Table 1 1 ) were analyzed by gas chromatography (Hewlett-Packard 58590 II Plus) coupled with a HP-5989 B mass spectrometer detector and equipped with column HP-5 (30 m×0.25 mm×0.025 µm). Temperature program: initial temperature, 70 °C; hold 2 min; temperature rate 4 °C/min, final temperature 250 °C; hold 30 min; column flow rate 0.8 ml He/min constant, injection temperature 250 °C; injection volume 1 µl/min. The mass spectrometry was accomplished with Wiley library ID 275, and setting as: electron impact ionization mode with 70 eV electron energy. Scan mass range m/z 45~50, detector temperature 250 °C. Essential oil Major constituents Abundance (%) 2 Caraway L-carvone 58.0 Limonene 38.4 3 Chenopodium p -cymene 33.3 Pinene-2-ol 11.7 Ascaridole 10.0 2 Cinnamon Cinnamaldehyde der. 36.1 Cinnamic aldehyde 12.3 2 Clove Eugenol 62.3 β-caryophyllene 19.2 3 Eucalyptus 1,8-cineole 45.1 Α-pinene 14.9 Α-terpineol 11.6 2 Garlic Pentdecane 12.1 Hexadecane 11.6 2 Geranium Citronellol 31.0 Citronellol formate 14.0 2 Lemon grass Limonene 91.1 Terpinene 2.2 3 Matrecary Camphor 51.4 p -cymene 20.1 Thymol 9.5 2 Peppermint Menthol 46.1 p -menthone 18.3 2 Rose Citronellol 34.4 Geranoil 20.4 2 Rosemary 1,8-cineole 28.6 Camphor 20.2 2 Thyme p -cymene 31.5 Thymol 18.5 3 Fennel Anthol 47.6 Pinene 11.7 1,8-cineole 10.3 Monoterpenoids 2 Fourteen monoterpenoidal constituents of the tested essential oils, namely: eugenol, (S & R) carvone, borneol, carveol, benzyl alcohol, thymol, camphor, menthol, chlorothymol, cinnamaldehyde, carvacrol, cineol, and citronellol and geraniol were supplied by Aldrich Chemical Company. The structures of the tested monoterpenoidal constituents are shown in Fig. 1 1 . Experimental treatments 2 Following preliminary experiments for either essential oil or monoterpenoid, five concentrations of 25×10 −6 , 50×10 −6 , 100×10 −6 , 250×10 −6 and 500×10 −6 per 0.5 g of dust and a control were selected. Each concentration was mixed with 0.5 g of house dust and placed on a microcell (3 cm×1.5 cm). Ten mites were introduced on top of the microcell with the aid a brush. Five replicates were maintained for each tested concentration. All experiments were conducted under laboratory conditions as the stock cultures were maintained. Mortality counts were done after 24 and 48 h of exposure. The LC 50 values and their confidence limits ( CL ) were computed according to Finney ( 1971 1971 ). RESULTS 1 The effect of fourteen plant essential oils namely: caraway, chenopodium, clove, cinnamon, eucalyptus, fennel, garlic, geranium, lemon, matrecary, peppermint, rose, rosemary and thyme, on the mortality of house dust mite, D . pteronyssinus (Trouessart), is presented in Table 2 2 . Five concentrations were used over time intervals 24 and 48 h under laboratory conditions. There was no mortality recorded in the controls. The 24 h- LC 50 results showed that clove oil was the most effective ( LC 50 =29.78×10 −6 ) followed by matrecary (104.45×10 −6 ), chenopodium (117.53×10 −6 ), fennel (156.42×10 −6 ) and caraway (158.05×10 −6 ). According to the GC-MS analysis (Table 1 1 ), clove oil contains 62.3% of eugenol and 19.2% of caryophyllene. Matrecary oil containing camphor 51.4%, p -cymene 20.1%, and thymol 9.5%, showed good activity after 24 h of the treatment with LC 50 value of 104.45×10 −6 . The result of the tested oils after 48 h showed that clove oil was the most effective followed by matrecary, chenopodium, rosemary and eucalyptus. From the obtained results, clove was the most promising oil tested against the house dust mite with the LC 50 values being 29.78×10 −6 and 21.17×10 −6 after 24 and 48 h, respectively. 2 Essential oil 2 Time (h) 2 LC 50 (×10 −6 ) 2 95% fiducial limits Lower Upper 2 Caraway 24 158.05 179.66 192.89 48 92.01 40.82 207.72 2 Chenopodium 24 117.53 97.23 142.19 48 76.02 53.71 160.10 2 Cinnamon 24 390.51 260.52 586.83 48 189.55 153.91 233.74 2 Clove 24 29.78 19.61 44.94 48 21.17 15.97 27.97 2 Eucalyptus 24 275.42 183.93 414.46 48 89.84 49.17 164.29 2 Fennel 24 156.42 122.40 185.09 48 94.46 81.42 109.61 2 Garlic 24 736.30 94.01 6054.90 48 571.31 271.73 1215.82 2 Geranium 24 181.53 146.07 225.93 48 118.56 62.37 226.61 2 Lemon 24 300.66 213.32 425.03 48 103.63 71.16 391.43 2 Matrecary 24 104.45 87.16 125.24 48 71.64 14.52 349.55 2 Peppermint 24 274.27 175.57 431.16 48 125.73 59.74 267.41 2 Rose 24 669.07 320.64 1405.82 48 409.28 160.34 1064.27 2 Rosemary 24 375.71 238.48 594.90 48 89.47 12.59 635.43 2 Thyme 24 488.65 259.86 919.04 48 269.59 216.46 335.96 The acaricidal activities of fourteen monoterpenoids against D . pteronyssinus were examined by direct application and are presented in Table 3 3 . Responses varied according to the compound tested and the time exposure. On the basis of LC 50 values, the compound most toxic against D . pteronyssinus was cinnamaldehyde (64.38×10 −6 ), followed by chlorothymol (90.45×10 −6 ), citronellol (159.48×10 −6 ), and menthol (253.68×10 −6 ). After 48 h, the obtained results of cinnamaldehyde and chlorothymol are comparable to each other with LC 50 values being 48.05×10 −6 and 45.81×10 −6 respectively. 2 Chemical 2 Time (h) 2 LC 50 (×10 −6 ) 2 95% fiducial limits Lower Upper 2 Carvacrol 24 318.33 157.69 652.63 48 115.48 86.49 154.50 2 Camphor 24 1026.58 353.78 3018.48 48 420.66 187.83 953.23 2 Citronellol 24 159.48 123.13 170.07 48 64.89 72.81 207.01 2 Carvone (R) 24 669.07 320.64 1405.82 48 253.13 118.33 547.46 2 Carvone (S) 24 643.89 72.81 6170.07 48 159.48 123.13 207.01 2 Carveol 24 309.36 272.15 437.58 48 85.37 72.01 101.15 2 Cineol 24 808.80 328.02 2023.69 48 488.05 259.86 919.04 2 Cinnamaldehyde 24 64.38 56.48 73.36 48 48.05 2.98 738.35 2 Chlorothymol 24 90.45 76.63 106.79 48 45.81 37.21 56.32 2 Borneol 24 638.65 272.14 1506.23 48 393.00 261.25 595.48 2 Benzyl alcohol 24 669.07 320.64 1405.82 48 181.79 114.12 270.20 2 Geraniol 24 431.96 58.03 3329.67 48 127.66 67.71 241.62 2 Menthol 24 253.68 73.06 907.36 48 136.89 24.69 779.96 2 Thymol 24 345.93 127.45 957.04 48 125.24 47.05 335.15 DISCUSSION 1 This study showed the effectiveness of some essential oils and their monoterpenoidal constituents against house dust mite D . pteronyssinus . Because of the strong activities of clove oil and some others, GC-MS was used to identify the essential oils components and their essential oil activity. Thymol and the structurally related compounds like p -cymene are effective as acaricides (Lee, 2004 2004 ). This observation is supported by the obtained results of chenopodium oil which also contains 33.3% of p -cymene. Matrecary oil exhibited high activity due to the presence of camphor 51.4%, p -cymene 20.1% and thymol 9.5%. Perhaps, there is a joint effect of those components leading to improved activity against mite. Rezk and Gadelhak ( 2004 2004 ) studied the effect of thymol and eugenol against the adult stage of the house dust mite, D. pteronyssinus . They showed that thymol and eugenol had almost the same effect against the tested mite. Interestingly, the similarity of the effect of rosemary and eucalyptus oils after 48 h may be due to 1,8-cineol as a major component which accounted for 28.6% and 45.1%, respectively (Table 1 1 ). The chlorinated compound of thymol (chlorothymol) was much more effective than thymol itself indicating the important role of chlorine atom on the miticidal activity. Much more interesting results emerged from the potent effect of cinnamaldehyde. Therefore, cinnamaldehyde and chlorothymol merit further study as potential dust mite control agents or as lead compounds. The public perception and acceptance of using washing product containing natural essential oils may be more positive. The most promising oil tested was clove which increased its potency over the test time. The toxic effect of these oils may be due to their fumigant and/or contact action. The vapor of some of these essential oils has been shown to kill pyroglyphidae mites (Watanabe et al., 1989 1989 ). Further screening of these essential oils in solution is required. Criteria for choosing the most suitable oil would include its acaricidal activity with time and temperature, volatility, the acceptability of fragrance, and adverse effects on fabrics. Essential oils may also be particularly suitable for inclusion in carpet washing solutions to aid in maintaining mite control. In conclusion, in our comprehensive screening, four essential oils, clove, matrecary, chenopodium and fennel and two monoterpenoids cinnamaldehyde and chlorothymol emerged as potential plant derived agents that provide promising results against house dust mites, D . pteronyssinus . The plant essential oils as a source for the control of the adult stage of house dust mite is a safe and effective approach, because many of them are selective to pests, with few if any harmful effects on non-target organisms and the environment (Susan and Ward, 1987 1987 ; Kim et al., 2003 2003 ; Kwon and Ahn, 2002 2002 ). 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