Larvicidal Effects of some Essential Oils against Larvae of the Pine Processionary Moth,
Thaumetopoea pityocampa (Denis & Schiffermüller) (Lepidoptera: Thaumetopoeidae)
K esdek, M.
*; N. Bayrak
**; S. Kordali
***; A. Usanmaz
***; G. Contuk
****and S. E rcisli
******Dept. of Environment Protection and Tecnologies)HWKL\H$OL6ÕWNÕ0HIKDUHW.RoPDQVocational School,
0X÷OD6ÕWNÕ.RoPDQ8QLYHUVLW\)HWKL\H0X÷OD-Turkey
** Dept. of Plant Protection, Agriculture Faculty, Bozok University, Yozgat-Turkey *** Dept. of Plant Protection, Agriculture Faculty, Atatürk University, 25240, Erzurum-Turkey **** Fethiye High School of Health Sciences, 0X÷OD6ÕWNÕ.RoPDQ8QLYHUVLW\)HWKL\H0X÷OD-Turkey
***** Dept. of Horticulture, Agriculture Faculty, Atatürk University, 25240, Erzurum-Turkey
(Received: June 9, 2013 and Accepted: July 21, 2013) A BST R A C T
The pine processionary moth, Taumetopoea pityocampa (Denis & Schiffermuller) (Lepidoptera: Thaumetopoeidae) is one of the most widespread defoliators found in the forest areas of Turkey. In this study, essential oils obtained from eight different plant species (Achillea gypsicola Hub-Mor., Origanum acutidens (Hand.-Mazz.) Ietswaart, Origanum onites L., Origanum rotundifolium Boiss., Satureja hortensis L., Satureja spicigera (C. Koch) Boiss., Tanacetum argyrophyllum (C. Koch) Tvzel. and Thymus sipyleus Boiss.) were tested against the 2nd, 3rd and 4th larval instars of T.
pityocampa under laboratory conditions. Results showed that the essential oils of all tested plantshad larvicidal effects on all the tested larval instars of T. pityocampa in comparison with control; mortality rates reached 100% in all thetested doses and exposure times in the positive controls.
K ey words: Essential oils, Larvicidal effect, Mortality percentage, Processionary moth, Taumetopoea pityocampa.
IN T R O DU C T I O N
The pine processionary moth, Thaumetopoea pityocampa (Den. & Schiff.) (Lepidoptera: Thaumetopoeidae) is one of the most important forest pests, especially in Mediterranean, Aegean and Marmara regions of Turkey. Larvae of this pest feed on Pinus species (Pinus brutia Ten., P. nigra Arnold, P. pinaster Aiton, P. pinea L., P. sylvestris
L.) and, also on Larix decidua Mill., Cedrus
atlantica Endl.and C. libani Rich. in forest areas of the world (Demolin, 1969 and Atakan, 1991). They cause serious economic and ecological losses on the host trees. Due to infestation, annual diameter increment of host trees decreases. This decrease has been reported to be from 12 to 65% (Babur, 2002; Hodar et al., 2002; Carus, 2004 and Kanat et al., 2005). Attacked trees can become highly prone to the incidence of secondary insects. Furthermore, these insects can cause death of tree in future $NNX]X DQG 6HOPL DQG $YFÕ DQG 2Jurlu, 2002). Therefore, the protection of coniferous forests requires regular applications of various control methods against this pest. These control means involve mechanical, physical, chemical, bio-technical and biological measures for such pest management. In the past, pesticides such as Endosülfan, Dimilin and Malathion had been used efficiently to control this pest in many countries. But, they had a negative effect on the environment and specifically on many beneficial organisms. Therefore, environmentally friendly methods are prefered (Roessler, 1989).
Many studies have been reported on the control the pine processionary moth in Turkey (Kanat and Sivrikaya, 2004; Özcankaya and Can, 2004 and Kanat and Mol, 2008). Pinus species (Pine trees) is one of the economically important plants for the forest villagers as wood and building material in Turkey. They are widespread along the coastal regions of Turkey, especially in the Mediterranean, Marmara, Aegean and Black Sea regions. Besides, they can grow in a latitude of 2000 meters.
Essential oils are natural plant products that contain natural flavours and fragrances grouped as monoterpenes (hydrocarbons and oxygenated derivatives), sesquiterpenes (hydrocarbons and oxygenated derivatives) and aliphatic compounds (alkanes, alkenes, ketones, aldehydes, acids and alcohols) that provide characteristic odours (Kordali
et al., 2007). Since the middle ages, essential oils have been widely used as bactericidal, virucidal, fungicidal, antiparasitical, insecticidal, medicinal and in cosmetic applications, especially nowadays in pharmaceutical, sanitary, cosmetic, agricultural and food industries. Because of the mode of extraction, which is mostly done by distillation from aromatic plants, they contain a variety of volatile molecules such as terpenes and terpenoids, phenol-derived aromatic components and aliphatic components (Bakkali et al., 2008). Essential oils of a large number of plant species have been found to have toxic and/or repellent effects against different insect species (Regnault-Roger, 1997). Recent investigations in several countries confirm that some
plant essential oils not only repel insects, but also have contact and fumigant insecticidal actions against some pests and fungicidal actions against some important plant pathogens (Isman, 2000). In recent years, a growing interest in research concerning the alternative pesticides and antimicrobial active compounds, including the plant extracts and essential oils that are relatively less damaging to the mammalian health and environment has been increased (Roy and Dureja, 1998; Isman, 2000; Cakir et al., 2004; Kordali et al., 2005: Emsen
et al., 2012 and Yildirim et al., 2012).
The genus Achillea is one of the most important genera of family Asteraceae and it comprises about 85 species, widespread throughout the world. There are about 42 species of this genus in Turkish flora and about 20 of them are endemic (Davis, 1982 and Baytop, 1999). The species of Achillea genus are NQRZQ LQ $QDWROLD DV ³&LYDQ SHUoHPL´ ³3LUHRWX´ DQG³<ÕODQoLoH÷L´
The genus Origanum (oregano) is an another important genus of family Lamiaceae and comprises about 900 species, widespread throughout the world. There are about 20 species of Origanum genus in Turkish flora (Davis, 1982 and Baytop, 1999).
Origanum species have traditionally been used as a spicy additive for food instead of thyme. This genus is rich in essential oils and bitter substances. The species of Origanum are known in Anatolia as ³<DODQFÕ NHNLN´ ³.HNLN´ ³øVWDQEXO NHNL÷L´ DQG ³.HNOLN RWX´ Origanum species are traditionally used as sedative, diuretic, degasifier, sweater and antiseptic, and also in the treatment of gastrointestinal diseases and constipation (Baytop, 1999).
Thymus sypleus, Satureja hortensis and S.
spicigera, that belong to family Lamiacaeae are well known aromatic and medicinal plants and are distributed in northern Anatolia. In Anatolia,
Thymus L. (thyme) and Satureja L. (savory) species are frequently used as tea or additives in commercial spice mixtures of many foods to offer aroma and flavour (Davis, 1982 and Baytop, 1999).
The genus Tanacetum is an important member of family Compositae. It is widespread in Europe and western Asia and consists of about 150±200 species. These species have traditionally been used as spicy additives for food, in cosmetics and as herbal remedies due to their biologically active compounds (Rohloff et al., 2004). This genus is represented by 44 species and altogether 59 taxa in Turkish flora. It is rich in essential oils, bitter substances and sesquiterpene lactones (Davis, 1982 and Baytop, 1999). Tanacetum species are known in Anatolia as ³SLUHRWX´ DQG WKHLU HVVHQWLDO RLOV DUH XVHG DV
repellent against insects (Baytop, 1999 and Baser
et al., 2001).
Turkish flora is characterized by abundance of aromatic plants among its components. The feature differentiates these plants from all others, in spite of the fact that they belong to different families, is the production of chemically related secondary compounds, the low molecular weight and volatile isoprenoids. This remarkable presence of aromatic species is important in determining the insectidal potential within this ecosystem.
Thus, the aim of this study was to evaluate possible totoxicity of the essential oils obtained from eight plants (A. gypsicola, O. acutidens, O. onites,
O. rotundifolium, S. hortensis, S. spicigera, T. argyrophyllum and T. sipyleus) from different localities of Turkey against the 2nd, 3rd and 4th larval instars of T. pityocampa under laboratory conditions.
M A T E RI A LS A ND M E T H O DS Insect Host
This study was conducted in the years 2011 and 2012. The 2nd, 3rd and 4th larval instars of T.
pityocampa were collected from infected Pinus brutia WUHHVLQWKHIRUHVWDUHDV .|\FH÷L]0X÷OD LQ Aegean Region of Turkey. Altitude of these forest areas is 150-250 m.
Bioassays
In order to test the toxicity of the essential oils against the 2nd, 3rd and 4th larval instars of T.
pityocampa, 10 larvae provided with appropriate amounts of fresh needles (P. brutia)were placed in Petri dishes (diameter 9 cm). Selected oil was dissolved in DMSO±water solution (10%, v/v). Final concentrations were 10 and 20 µl/ml. Emulsions were spread by a glass atomizer to Petri dish (9 cm diameter), placed on the bottom at two layers of filter paper (1 ml/Petri dishes). This study was carried out under the laboratory conditions (23±1°C, 60±5% RH and a photoperiod of 8:16 h (L: D)). At different exposure times (24, 48, 96 hours), larval mortality was determined. A Petri dish, applied with only sterile water and DMSO, was used as control. In addition, Kormilin 25 WP (10 and 20µl/Petri dish) was used as positive control. Three replicates were used for each dose and exposure time combination. The larvicidal activity of the essential oils of these eight plants was expressed as % mean mortality of the larvae.
Plant material and isolation of essential oil
A. gypsicola, O. acutidens, O. onites, O. rotundifolium, S. hortensis, S. spicigera, T. argyrophyllum and T. sipyleus were collected at flowering stage from different localities of Turkey in
2011. Collected plant materials were dried in shadow and ground in a grinder. The dried plant samples (500g) were subjected to hydrodistillation (plant material in boiling water), using a Clevenger-type apparatus for 4 h. Hydrodistillation of A. gypsicola, O. acutidens, O. onites, O. rotundifolium,
S. hortensis, S. spicigera, T. argyrophyllum and T. sipyleus yielded 0.65, 0.60, 3.67, 1.23, 2.3, 1.56, 0.75 and 0.98 % (w/w) of essential oils, respectively. The yields were based on dry materials of plant samples. The essential oils were stored in a freezer at 4ºC for further tests.
Statistical analysis
Differences among fumigant activities of essential oils tested were determined according to analysis of variance (ANOVA) test by using SPSS 13.0 software package. Differences between means were tested through Duncan tests and values with p<0.01 and p<0.05 were considered significantly different.
R ESU L TS A ND DISC USSI O N
Toxicity effects of the essential oils extracted from A. gypsicola, O. acutidens, O. onites, O.
rotundifolium, S. hortensis, S. spicigera, T. argyrophyllum and T. sipyleus on the 2nd, 3rd and 4th larvae instars of T. pityocampa are summarized in (Tables 1, 2 and 3). The results show that essential oils of A. gypsicola, O. acutidens, O. onites, O. rotundifolium, S. hortensis, S. spicigera, T. argyrophyllum and T. sipyleus had larvicidal effects on 2nd, 3rd and 4th larvae instars of T. pityocampa in comparison with the control. Minimum mortality rate (80%) was obtained, 24 h post treatment at the ȝOHVVHQWLDORLORIA. gypsicola for the 2nd larval instar of T. pityocampa 8VLQJ ȝO RI GLIIHUHQt tested essential oils gave 100% mortality after 24 h against the 2nd larval instar, with an exception of the oils extracted from S. hortensis and A. gypsicola,
which gave 95.6 and 90.0% mortality excessively using.
Minimum mortality rate (60%) was recorded 24 h post using essential oil of S. spicigera on the 3rd larval instar of T. pityocampa DWȝO . In addition, mortality rates were 66,6 and 73,3%, respectively DIWHU DQG KV LQ WKH ȝO However, the mortality rates were 100% in all other plants for all doses after 96 h (Table 2).
The least mortality rate (46,6%) was recorded 24 KSRVWWUHDWPHQWDWWKHGRVHRIȝORIHVVHQWLDORLO of S. spicigera on the 4th larval instar of T.
pityocampa. The minimum mortality rates were found 24 h post treatment, using different doses (10 ȝODQGȝO RIHVVHQWLDORLOVRIDOOSODQWVRQthe 4th larval instar (Table 3). However, the mortality rates
were determined as (100%) in all doses of all the essential oils for the 2nd, 3rd and 4th larval instars of
T. pityocampa 96 h post treatment (Tables 1, 2 and 3). The least mortality rates 46,6, 66,6 and 73,3%, respectively were obtained 24, 48 and 72 hs post WUHDWPHQWXVLQJȝOHVVHQWLDORLORIS. spicigera in comparison with other essential oils on the 4th larval instar (Table 3). However, there was no mortality in the control during the tested period. In addition, mortality rate was found to be 100% at all doses and exposure times in the control.
On the other hand, mortality rates increased among all larval instars with increasing doses and exposure times of the essential oils of all the tested plants. The essential oil of O. rotundifolium had the highest larvicidal effect (100%) among all exposure times and treatment doses on all larval instars (Tables 1, 2 and 3).
It has been previously stated that the essential oil isolated from O. acutidens had a toxic effect on
Sitophilus granarius and Tribolium confusum
(Kordali et al., 2008). As well, the essential oil of O.
onites had also a toxic effect on 4th and 5th larval instars of T. pityocampa (Çetin et al., 2006). In addition, the essential oil of O. onites had 100% mortality rate against larval instars of Ephestia
kuehniella and Pilodia interpunctella (Ayvaz et al., 2010). In the present study, the essential oil of O. onites had a larvicidal effect (100% mortalty) at all exposure times and treatment doses (except 24 h on 3rd and 4th larval instars, at ȝOGRVH
The essential oil of O. rotundifolium had an insecticidal effect on adults of S. granarius
<ÕOGÕUÕPet al., 2011). The present study proved that the essential oil of O. rotundifolium had a larvicidal effect in all exposure times and treatment doses with 100% mortality rate on all larval instars of T. pityocampa.
In an earlier study, it was reported that a test for the toxicity of S. hortensis oil showed an insecticidal effect against Bruchus dentipes and the mortality increased with increasing doses and exposure times for the essential oil (Tozlu et al., 2011). On the other hand, the essential oil of S. hortensis had insecticidal activities (fumigant, repellent and contact toxicity) against T. castaneum, E. kuehniella and P. interpunctella (Maedeh et al., 2011). However, it was determined that the essential oil extracted from
S. hortensis had insecticidal activity against male and female adults of C. maculatus (Heydarzade and Moravvej, 2012).
The essential oil isoleted from S. spicigera
<ÕOGÕUÕP et al., 2011) and T. argyrophyllum
Table (1): Mortality rate of the 2nd larval instar of T. pityocampa when treated with different doses of different plant essential oils at exposure time intervals
Essential oils Exposure time (h) Mortality(%)
Plants species Dose (µl/l) 24 48 72 96
Satureja hortensis 10 96.6 ± 5.77 c 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Thymus sipyleus 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Achillea gypsicola 10 90.0 ± 0.0 c 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 80.0 ± 17.3 b 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Satureja spicigera 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Origanumonites 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Origanumrotundifolium 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Tanacetum argyrophyllum 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Origanum acutidens 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Positive control (Kormilin 25 WP) 10 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 b 100 ± 0.0 b 100 ± 0.0 b Control (DMSO+Su) - 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a
9DOXHV IROORZHG E\ GLIIHUHQW OHWWHUV LQ WKH VDPH FROXPQ GLIIHU VLJQLILFDQWO\ DW 3 DFFRUGLQJ WR 'XQFDQ 0XOWLSOH test.
a Mean±SE of three replicates, each set up with 10 adults.
Table (2): Mortality rate of the 3rd larval instar of T. pityocampa when treated with different doses of different plant essential oils at exposure time intervals
Essential oils Mortality (%)
Exposure time (h)
Plant species Dose (µl/l) 24 48 72 96
Satureja hortensis 10 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Thymus sipyleus 10 96.6 ± 5.77 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Achillea gypsicola 10 63.3 ± 5.77 b 96.6 ± 5.77 b 100 ± 0.0 c 100 ± 0.0 b 20 76.6 ± 15.2 c 100 ± 0.0 b 100 ± 0.0 c 100 ± 0.0 b Satureja spicigera 10 60.0 ± 10.0 b 66.6 ± 5.77 b 73.3 ± 11.5 b 100 ± 0.0 b 20 96.6 ± 5.77 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Origanumonites 10 96.6 ± 5.77 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Origanumrotundifolium 10 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Tanacetum argyrophyllum 10 90.0 ± 10.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Origanum acutidens 10 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Positive control (Kormilin 25 WP) 10 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 d 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Control (DMSO+Su) - 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a
9DOXHV IROORZHGE\GLIIHUHQW OHWWHUVLQ WKHVDPH FROXPQGLIIHUVLJQLILFDQWO\DW 3DFFRUGLQJWR Duncan Multiple test.
Table (3): Mortality rate of the 4th larval instar of T. pityocampa when treated with different doses of different plant essential oils at exposure time intervals
Essential oils Exposure time (h) Mortality (%)
Plant species Dose (µl/l) 24 48 72 96
Satureja hortensis 10 93.3 ± 11.5 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Thymus sipyleus 10 90.0 ± 17.3 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 90.0 ± 17.3 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Achillea gypsicola 10 80.0 ± 20.0 c 96.6 ± 5.77 b 100 ± 0.0 c 100 ± 0.0 b 20 83.3 ± 15.2 c 100 ± 0.0 b 100 ± 0.0 c 100 ± 0.0 b Satureja spicigera 10 46.6 ± 20.8 ab 66.6 ± 20.1 b 73.3 ± 15.2 b 100 ± 0.0 b 20 93.3 ± 5.77 c 96.6 ± 5.77 c 96.6 ± 5.77 c 100 ± 0.0 b Origanumonites 10 80.0 ± 5.77 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Origanumrotundifolium 10 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Tanacetum argyrophyllum 10 80.0 ± 17.3 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Origanum acutidens 10 50.0 ± 20.0 b 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 20 80.0 ± 20.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b Positive control (Kormilin 25 WP) 10 20 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 c 100 ± 0.0 b 100 ± 0.0 b Control (DMSO+Su) - 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a 0.0 a ± 0.0 a
9DOXHV IROORZHGE\GLIIHUHQW OHWWHUVLQ WKHVDPH FROXPQGLIIHUVLJQLILFDQWO\DW 3DFFRUGLQJWR 'XQFDQ0XOWLSOH test.
a Mean±SE of three replicates, each set up with 10 adults. adults of S. granarius. In the present study, the essential oil of S. spicigera had 100% mortality at all exposure times (24, 48, 72 and 96 hs) and treatment doses ( ȝO DQG ȝO on 2nd larval instar of T.
pityocampa and 90 and 80%(24 h, ȝOGRVH RQ3rd and 4th larval instars, respectively.
The essential oil of T. sipyleus had an insecticidal effect against adults of S. granarius. High doses and long exposure times resulted to maximum toxicity on
S. granarius <ÕOGÕUÕPet al., 2011). The essential oil of T. sipyleus caused 100% mortality at all exposure times and treatment doses on all larvae instars of T. pityocampa except 24 h, ȝO GRVH RQ 3rd larval instar, 24 h, ȝODQGȝOGRVHVon 4th larval instar that caused 96,6 90 and 90%, respectively.
Obtained results agree with some previous studies which demonstrated that the essential oil of A. gypsicola had potent toxic effect against adults of B. dentipes and some other insect species (Sampson et al dDOPDúXUet al., 2006 and Tozlu et al., 2011).
In conclusion, results of the present study demonstrated that the larvicidal effect of essential oils obtained from eight different plant species, especially that of O.rotundifolium, was considerable. Therefore, these essential oils can be recommended as potential alternatives to control larval instars of T.
pityocampa. However, further field research is
needed in order to suppress its damage on the forest trees.
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