Evaluation of Tung Oil (Vernicia fordii (Hemsl.)) for Controlling Termites

Abstract

In worldwide, the use of chemical pesticides to protect wood has been greatly restricted. In recent years, a large number of researchers devoted to the search for natural, safe and non-polluting bioactive chemical compounds from plants as an alternative to synthetic organic chemical preservative. In Chinese folk, tung oil can be used as paint for wooden furniture to protect them from pests. This study aimed to evaluate the chemical compositions of raw and heated tung oil and their activity against termite. In choice bioassays, weight loss of wood treated with 5% raw or heated tung oil after 4 weeks was significantly less than that of the control group. In no-choice bioassays, there was a significant difference in termite survival and wood weight loss on raw and heated tung oil-treated wood. When tung oil-treatment concentrations increased to 5%, wood weight loss was less than 10%. There was no significant difference in termite survival and wood weight loss between raw and heated tung oil-treated wood. Survival of termites in both tung oil wood treatments was significantly lower than that in the starvation control after 4 weeks. Raw and heated tung oil significantly improved the resistance of pine wood to termites, and have the potential for the development of natural wood preservatives.

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15
Journal of Botanical Research | Volume 04 | Issue 03 | July 2022
Journal of Botanical Research
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Copyright © 2022 by the author(s). Published by Bilingual Publishing Co. This is an open access article under the Creative Commons
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*Corresponding Author:
Yongjian Xie,
College of Advanced Agricultural Sciences, Zhejiang A&F University, Hangzhou 311300, China;
Email: yjxie@zafu.edu.cn
DOI: https://doi.org/10.30564/jbr.v4i3.4793
ARTICLE
Evaluation of Tung Oil (Vernicia fordii (Hemsl.)) for Controlling
Termites
Hangtian Li1 Siying Li1 Hui Lu2 Jingjing Zhang1 Xi Yang2 Dayu Zhang2 Yike Zhang2
Yongjian Xie2*
1. Institute of Termite Control of Yuhang, Hangzhou, 311100, China
2. College of Advanced Agricultural Sciences, Zhejiang A&F University, Hangzhou 311300, China
ARTICLE INFO ABSTRACT
Article history
Received: 14 June 2022
Revised: 12 July 2022
Accepted: 13 July 2022
Published Online: 31 July 2022
In worldwide, the use of chemical pesticides to protect wood has been
greatly restricted. In recent years, a large number of researchers devoted
to the search for natural, safe and non-polluting bioactive chemical
compounds from plants as an alternative to synthetic organic chemical
preservative. In Chinese folk, tung oil can be used as paint for wooden
furniture to protect them from pests. This study aimed to evaluate the
chemical compositions of raw and heated tung oil and their activity against
termite. In choice bioassays, weight loss of wood treated with 5% raw or
heated tung oil after 4 weeks was signicantly less than that of the control
group. In no-choice bioassays, there was a signicant difference in termite
survival and wood weight loss on raw and heated tung oil-treated wood.
When tung oil-treatment concentrations increased to 5%, wood weight loss
was less than 10%. There was no signicant difference in termite survival
and wood weight loss between raw and heated tung oil-treated wood.
Survival of termites in both tung oil wood treatments was significantly
lower than that in the starvation control after 4 weeks. Raw and heated tung
oil signicantly improved the resistance of pine wood to termites, and have
the potential for the development of natural wood preservatives.
Keywords:
Termite-resistance
Raw and heated tung oil
Vernicia fordii
Coptotermes formosanus
1. Introduction
Coptotermes formosanus (Shiraki) is an economically
important insect pest that damages wooden structures
[1]. Rust and Su [2] reported that the worldwide economic
loss caused by this species is at least $40 billion annual-
ly. Ammonium copper quaternary (ACQ), copper citrate
(CC), and copper chrome arsenic (CCA) have been used
to treat wood and prevent termite damage, but these com-
pounds can affect human health and the environment [3].
Therefore, there is interest in more environment-friendly,
convenient, and effective wood preservatives. Some wood

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Journal of Botanical Research | Volume 04 | Issue 03 | July 2022
is naturally resistant to termites [1,4], due to their active
compounds that act as toxicants [2,5]. These plant compounds
include tannins [4], avonoids [6,7], alkaloids [8,9], quinones [10-12],
terpinoids [13-15], and resins [16-18]. Many studies have docu-
mented that active compounds possess antitermitic prop-
erties [13,14,19].
Tung tree, Vernicia fordii (Hemsl), native to China,
is widely cultivated in China and other countries for its
industrial values [20,21]. V. fordii is mainly distributed in Si-
chuan, Hubei, Guizhou, and Hunan provinces and Chong-
qing municipality, which is the main production base of
tung oil in China [22,23]. Tung oil, extracted from seeds,
shows rapid drying, insulation, anticorrosion, acid, and
alkali resistance [24]. These properties make tung oil a val-
uable additive in paints, varnishes, and other coatings and
nishes [25]. In addition, tung tree seeds, roots, owers, and
leaves are widely used in folk medicine. Tung oil can treat
burns, scalds, and cold injuries [26]. Termites are important
pests of wooden structures. Many essential oils have anti-
termitic activities, including oils from Lippia spp. [27], Ar-
temisia absinthium [28], Lippia sidoides [29], Chamaecyparis
formosensis [30], Liquidambar orientalis, and Valeriana
wallichii [31]. Hutchins [32] reported that Aleurites fordii
wood and meal extracts had antitermitic toxicity. Tung
seed is considered a good wood preservative in Chinese
and is used to treat indoor and outdoor wooden furniture
to protect them against insect pests and wood-rot fungi.
However, the protection against termite damage offered
by tung oil treatment of wood is unclear. Therefore, the
purpose of this study was to compare the chemical com-
position of raw and heated tung oil and to determine the
toxicity of raw and heated tung oil to termites.
2. Materials and Methods
2.1 Chemicals
The fatty acid methyl ester standard was obtained from
Dr. Ehrenstorfer GmbH. Methanol and isooctane were
purchased from Sigma Aldrich Inc, St. Louis, MO, USA.
Potassium hydroxide and sodium bisulfate were obtained
from Sinopharm Chemical Reagent Co., Ltd (Beijing,
China).
2.2 Tung Oil
Tung oil can be divided into raw tung oil and heated
tung oil. Raw tung oil was obtained from tung seeds using
a hydraulic press at room temperature, with characters of
golden yellow and strong penetration. Heated tung oil was
made by heating the raw tung oil to 180 °C, brown color,
low transparency, and high density. The oils were supplied
by the Yiyousheng home exclusive store, Tmall.com. The
colors of the raw and heated tung oil were light yellow
brown and dark brown, respectively.
2.3 Esterication of Tung Oil Fatty Acids
The esterication of tung oil fatty acid was pretreated
according to ISO 5509-2000. A 0.010 g oil sample and 1
mL of 1 M KOH-methanol solution were added to a round
bottom flask and heated in a water bath at 75 °C for 10
min. After cooling to room temperature, 1 mL of C15:0
was added to the mixture. After transfer to a 250 mL sep-
arating funnel, 20 mL of n-heptane and 20 mL of water
were added to stratify the mixture, and then we collected
the ester layer and dried it with anhydrous sodium sulfate.
2.4 Gas Chromatography-mass Spectrometry
(GC-MS)
GC-MS analysis was performed on a gas chromato-
graph Agilent 6890A interfaced with an Agilent 5975C
mass spectrometer (Agilent Technologies (China) Co.,
Ltd.). An HP-5 MS capillary column (30 m × 0.25 mm ×
0.25 μm) was used. The column temperature was pro-
grammed to rise from 50 °C to 280 °C at a rate of 10
°C/min. The carrier gas was helium with a flow rate of
1 mL/min. MS readings were taken at 70 eV and a mass
range of 15-500. Identication of compounds of the tung
oil was based on standard samples, and NIST11.LIB (Na-
tional Institute of Standards and Technology) was used for
qualitative analysis.
2.5 Oil Treatment
The vacuum-soak impregnation method by Nakay-
ama and Osbrink [33] was used with slight modications.
Masson pine (Pinus massoniana) was cut into wood
pieces that were 23 mm (length) × 14 mm (width) × 9
mm (thickness). These were sterilized in an oven at 130
°C for 24 h and weighed, and the weights were recorded.
The wood was placed into a closed pressure chamber with
vacuum level of –0.098 MPa for 30 min. Then, the wood
specimens were immersed with different concentrations
of both raw and heated tung oil-acetone mixtures, soaked
for 30 min, and removed. Excess liquid on the wood sur-
face was dried with paper, and the blocks were dried in a
vacuum oven at 60 °C for 24 h. This procedure was used
to remove the acetone fraction. The oil content of the
oil-treated wood (%, w/w) was determined based on the
gain in weight of the untreated wood.

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2.6 Termites
The colonies of Coptotermes formosanus Shiraki were
collected from Tiantong Mountain in Ningbo, Zhejiang
province. The termites were reared on masson pine blocks
(200 mm ×35 mm×20 mm), water soaked in plastic con-
tainers (460 mm × 36 mm × 28 mm) at 26 ± 1 °C and 80 ±
5% RH. Termites were identified using keys for soldier
identication from Scheffrahn and Su [34].
2.7 Termite Resistance Test
The choice and no-choice bioassay methods of Nakay-
ama and Osbrink [33] were used to evaluate termite resist-
ance of pine wood treated with tung oil.
2.7.1 Choice Bioassays
The wood specimens tested contained raw and pro-
cessed tung oil contents of 1.25%, 2.5%, 5.0%, 10.0%,
20.0%, and 40.0% (w/w). The termite control group was
exposed to pine treated only with acetone. Two blocks
of wood were placed on a sand surface on both sides of
the bottom of the container (65 mm (diameter) × 90 mm
(high)) (Figure 1). A total of 150 workers and 5 soldiers were
placed in the container. Three replicates of the oil treatments
were performed. All the testing containers were placed in a
conditioning room at 26 ± 1 °C, and after 4 weeks, the wood
specimens were dried and reweighed to determine wood
weight loss. ASTM [35] ratings were determined for each
wood block over the same period. The ASTM rating had a
scale of 10-0 with 10 being sound wood with only surface
nibbles permitted, 9 light attack, 7 moderate attack with pen-
etration, 4 heavy attack, and 0 failure.
Figure 1. A test unit for evaluating termite resistance to
wood treated in the laboratory. A: No-choice; B: Choice
2.7.2 No-choice Bioassays
The experimental method was the same as above, but
only one piece of treated wood was placed in the middle
of the container.
2.8 Statistical Analysis
For choice bioassays, Student’s t test was used to com-
pare wood specimen weight loss of untreated with treated
wood specimens after 4 weeks. Differences in survival,
wood specimen weight loss, and percent weight loss of
wood were compared using ANOVA followed by the Stu-
dent-Newman-Keuls test (p < 0.05).
3. Results
3.1 Chemical Compositions of Tung Oil
GC-MS analysis results of the raw and heated tung oil
are shown in Table 1. The main components of raw tung
oil were ɑ-eleostearic acid (69.02%), oleic acid (12.28%),
and linoleic acid (8.93%). ɑ-eleostearic acid (65.31%),
oleic acid (15.52%), and linoleic acid (9.44%) were the
major components of the heated tung oil.
Table 1. Chemical composition (%) of raw and heated
tung oil
No. RIaCompounds Raw Heated
1
2
3
4
5
1973
2123
2134
2179
2482
Palmitic acid
Linoleic acid
Oleic acid
Stearic Acid
ɑ-eleostearic acid
3.24
8.93
12.28
2.58
69.02
4.16
9.44
15.52
3.27
65.31
aRI, retention index calculated on the HP-5MS column relative
to C8-C28 n-alkanes.
3.2 Choice Bioassays
There was significantly less wood weight loss of the
blocks with 5% raw and heated tung oil after 4 weeks
(Table 2) than in the untreated blocks. There was high ter-
mite survival in both tung oil treatments at 4 weeks. There
were signicant differences in termite survival and wood
weight loss (F = 5.406, df = 6, 14, P < 0.004; F = 16.374,
df = 6, 14, P < 0.0001) on raw and heated tung oil-treated
wood, respectively. These results were corroborated using
the ASTM rating system (Table 3).

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Journal of Botanical Research | Volume 04 | Issue 03 | July 2022
3.3 No-choice Bioassays
There were significant differences in termite survival
(F = 72.87; df = 6, 14, P < 0.0001; F = 39.83, df = 1, 97,
P < 0.0001) and wood weight loss (F = 44.32, df = 4, 16,
P < 0.0001; df = 4, 16, F = 50.97, P < 0.0001) on raw and
heated tung oil-treated wood, respectively. Survival and
wood weight loss in both tung oil-treatments decreased as
the concentration increased (Table 4). When the raw and
treated tung oil concentrations increased to 5.0%, percent
weight loss of wood was less than 10% (Table 4). ASTM
ratings were highest when both raw and heated tung oil
content was 40% (Table 5). There were no significant
differences in termite survival (F = 0.257; df = 5, 36; P =
0.932) and wood specimen weight loss (F = 0.403; df = 5,
36; P = 0.842) on raw and heated tung oil-treated wood.
Table 2. Comparison of weight loss between wood specimens treated with both raw and heated tung oil 4 weeks after
exposure to C. formosanus in the feeding choice bioassays.
Treatment
% (w/w) Survival, %
Weight loss (mg) and t statistics (mean ± SD)
Treated Untreated t P
raw tung oil
40
20
10
5
2.5
1.25
heated tung oil
40
20
10
5
2.5
1.25
Control
Acetone
Untreated
78.0 ± 3.1d
89.8 ± 5.7ab
81.6 ± 3.0cd
81.1 ± 3.8cd
82.2 ± 2.7cd
90.4 ± 2.3ab
80.2 ± 1.1cd
87.8 ± 2.0abc
82.0 ± 2.4cd
93.8 ± 1.7a
81.3 ± 2.4cd
90.7 ± 2.9ab
85.3 ± 3.1bcd
84.7 ± 2.4bcd
36.7 ± 29.1
61.3 ± 27.8
85.1 ± 34.0
102.0 ± 60.9
191.5 ± 50.7
278.2 ± 62.2
33.8 ± 11.2
50.1 ± 12.1
55.8 ± 21.5
73.8 ± 18.0
206.2 ± 55.1
237.7 ± 28.7
259.4 ± 32.1
212.5 ± 26.9
271.8 ± 37.2
249.4 ± 38.3
247.3 ± 19.7
243.1 ± 23.1
221.1 ± 58.9
236.3 ± 18.6
298.5 ± 23.9
321.0 ± 67.7
279.2 ± 64.3
271.7 ± 54.8
242.2 ± 40.3
257.5 ± 52.0
253.0 ± 35.8
242.5 ± 107.9
8.63
6.89
7.16
3.75
0.66
1.12
17.34
6.83
5.70
5.94
0.91
0.58
0.23
0.47
0.001
0.002
0.002
0.02
0.55
0.33
0.0001
0.002
0.005
0.004
0.41
0.60
0.83
0.67
Table 3. Comparison of ASTM ratings between wood specimens treated with both raw and heated tung oil and untreat-
ed wood specimens 4 weeks after exposure to C. formosanus in a feeding choice bioassay.
Treatment
% (w/w)
ASTM rating of wood and t statistics (mean ± SD)a
Treated Untreated t P
raw tung oil
40
20
10
5
2.5
1.25
heated tung oil
40
20
10
5
2.5
1.25
Control
Acetone
Untreated
9.7 ± 0.6
9.3 ± 0.6
8.8 ± 0.3
7.7 ± 1.2
4.7 ± 1.2
4.0 ± 1.0
9.8 ± 0.3
9.5 ± 0.5
9.3 ± 0.0
8.7 ± 1.5
4.0 ± 1.0
5.0 ± 1.7
4.3 ± 0.6
4.3 ± 1.5
0.0 ± 0.0
0.0 ± 0.0
0.0 ± 0.0
0.0 ± 0.0
3.7 ± 1.5
3.3 ± 0.6
0.0 ± 0.0
0.0 ± 0.0
0.0 ± 0.0
0.0 ± 0.0
3.7 ± 1.2
4.0 ± 1.5
5.0 ± 1.7
4.6 ± 2.1
29.00
28.00
53.00
11.5
0.655
1.09
59.00
32.91
28.00
9.827
1.09
2.01
0.555
0.229
0.001
0.001
0.0001
0.007
0.58
0.423
0.0001
0.001
0.001
0.01
0.423
0.184
0.635
0.84
a ASTM scale of 10-0 with 10 being sound, surface nibbles permitted, 9 having light attack, 7 moderate attack with penetration, 4
with heavy attack, and 0 failure.

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Journal of Botanical Research | Volume 04 | Issue 03 | July 2022
4. Discussion
Biodegradation of wood by fungi and termites is a
serious problem for wooden structures world-wide [15].
The alkaline copper quat (ACQ) [36], boron-uorine-chro-
mium-arsenic (BFCA salts) [37], copper azole (CA) [38],
copper chrome arsenate (CCA) [39], chlorotalonil (CTL) [36],
copper- and zinc-salicylate [40], quaternary ammonium com-
pounds (QACs) [41], siloxane [42], sodium uoride (NaF) [43-45],
and zinc borate [46] have been used to protect wood against
termite damage. Besides these, nanoparticles from zinc
oxide (ZnO) [47,48], CuO and B2O3 [49], and magnesium uo-
ride (MgF2) [50] have provided promising levels of protec-
tion. Wood preservatives should ideally be environmental-
ly friendly; thus, there is interest in safer alternative wood
protection methods. Secondary plant compounds in some
species of wood play a major role in the protection of
wood against termite attack [51-55]. Here, we evaluated the
chemical composition of tung oil and tested their activity
against termite.
The major components of tung oil studied in this paper
are similar to previous reports on this species. Raw tung
oil, extracted from seeds, contains 60%~80% α-eleostear-
ic acid and is used for the production of biodiesel, dyes,
inks, and resins [20,56,57]. The α-eleostearic acid has a po-
Table 4. Comparison of termite survival and wood specimens weight loss between wood specimens treated with both
raw and heated tung oil 4 weeks after exposure to C. formosanus in the feeding no-choice bioassays.
Treatment
% (w/w)
mean ± SDa
Survival, % Total wt loss, mg Percent weight loss, %
raw tung oil
40
20
10
5
2.5
1.25
heated tung oil
40
20
10
5
2.5
1.25
Control
Acetone
Untreated
No food
26.7 ± 2.4e
45.1 ± 2.1d
47.3 ± 1.3cd
48.7 ± 6.6cd
55.3 ± 2.7bcd
63.1 ± 5.2b
27.8 ± 2.0e
49.1 ± 5.6cd
51.8 ± 6.0cd
55.3 ± 2.7bcd
58.4± 2.8bc
65.8 ± 8.4b
84.7 ± 4.8a
86.2 ± 2.8a
83.1 ± 5.4a
36.5 ± 4.5d
96.5 ± 20.5bc
138.8 ± 15.0b
168.4 ± 28.4b
256.1 ± 29.6a
281.7 ± 22.8a
54.3 ± 23.8cd
105.8 ± 12.4bc
141.4 ± 6.1b
163.4 ± 9.6b
280.9 ± 21.2a
294.7 ± 30.0a
284.4 ± 67.0a
312.2 ± 45.7a
-
1.9 ± 0.2d
5.4 ± 1.1cd
6.9 ± 0.8cd
10.2 ± 1.7c
17.3 ± 2.0b
17.0 ± 1.4b
2.6 ± 1.2d
6.1 ± 0.7cd
8.4 ± 0.4c
8.9 ± 0.5c
18.2 ± 1.4b
23.5 ± 2.4a
24.5 ± 5.7a
24.6 ± 3.6a
-
a Means ± SD followed by the different letter within a column are signicantly different (P < 0.05; using Student-Newman-Keuls
test).
Table 5. Comparison of ASTM ratings between wood specimens treated with both raw and heated tung oil and untreat-
ed wood specimens 4 weeks after exposure to C. formosanus in a feeding no-choice bioassay.
Treatment % (w/w) ASTM rating of wood (mean ± SD)a
raw tung oil heated tung oil
40
20
10
5
2.5
1.25
Acetone
Untreated
9.7 ± 0.6a
8.3 ± 1.5ab
6.7 ± 2.5b
4.6 ± 0.6c
0.0 ± 0.0d
0.0 ± 0.0d
0.0 ± 0.0d
0.0 ± 0.0d
9.3 ± 0.6A
8.0 ± 1.0B
6.3 ± 0.6C
4.3 ± 1.5D
0.0 ± 0.0E
0.0 ± 0.0E
0.0 ± 0.0E
0.0 ± 0.0E
a ASTM scale of 10-0 with 10 being sound, surface nibbles permitted, 9 having light attack, 7 moderate attack with penetration, 4
with heavy attack, and 0 failure.

20
Journal of Botanical Research | Volume 04 | Issue 03 | July 2022
tential role in human health products such as those with
antibacterial [58], antitumor [59], anti-neuroinammatory [26],
antioxidative [60], and antiobesity [61] activity. It also pro-
vides a reference on the safety of tung oil used in commer-
cial applications. In addition, ɑ-eleostearic acid can prevent
Anthonomus grandis damage to cotton bolls [62]. Thus, high
anti-termitic properties of tung oil could be due to the
high toxicity of its major component ɑ-eleostearic acid.
Similarly, Xie et al. [15] showed that the major constituents
of Syzgium aromaticum, eugenol, against Reticulitermes
chinensis for 1 d, 3 d and 5 d had LC50 values of 38.0 μg/g,
12.1 μg/g, and 9.2 μg/g, respectively. Yang et al. [63] also
demonstrated that the high toxicity of Mentha spicata EO
against Reticulitermes dabieshanensis was attributed to
its major components, carvone, limonene and dihydro-
carvone.
In choice experiments, there was no signicant differ-
ence in termite survival and wood weight loss between
raw and heated tung oil-treated wood. Therefore, raw tung
oil can be directly used for wood preservation to reduce
the processing cost of heated tung oil. Under starvation
conditions for 4 weeks, termite survival was signicantly
higher than that of raw and heated tung oil-treated wood,
indicating that raw and heated tung oil were feeding de-
terrents for C. formosanus. Similar results were obtained
showing that tung wood and meal have antitermitic prop-
erties [32]. In addition, Nakayama and Osbrink [33] showed
that A. moluccana oil-treated wood was resistant to C.
formosanus. Taylor et al. [64] observed that removal of the
methanol-soluble heartwood components of Thuja plicata
and Chamaecyparis nootkatensis reduced their resistance
to termite attack. Syofund et al. [65] observed that wood
extracts used as preservatives improved the resistance of
less durable wood to termite attack by 50% compared to
the controls. Brocco et al. [66] found that ethanol extracts
of Tectona grandis heartwood increased the resistance and
mortality against Nasutitermes corniger in both choice
and no-choice tests. Similar results were reported by
Hassan et al. [67], who showed that Tectona grandis and
Cedrus deodara extracts imparted termite resistance to
non-durable wood species.
5. Conclusions
In this study, we investigated the resistance of pine
wood (P. massoniana), treated with raw and heated tung
oil, to the termite, C. formosanus. Our study demonstrat-
ed that both tung oil treatments significantly improved
the resistance of pine wood to termites. When both tung
oil-treated concentrations were 5.0%, the weight loss of
wood was less than 10%. There was no significant dif-
ference in termite survival rate and wood weight loss be-
tween the two tung oil treatments. Therefore, raw tung oil
can be directly used for wood preservation to reduce the
processing cost of heated tung oil. In addition, the use of
tung oil for wood preservative treatment can expand the
demand for tung oil and increase local economic income.
Acknowledgments
This study was funded by College Students’ Scientic
Research Training Program of Zhejiang A&F University
(No. 113-2013200148) and ZAFU-Institute of Termite
Control of Yuhang cooperative project (2045200485,
2045200529).
Conict of Interest
There is no conict of interest.
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