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Exterminating Effect of Wood Vinegar to Red Mites and
its Safety to Chickens
Kohsyo Yamauchi1, Noboru Manabe
2, Yoshiki Matsumoto
3and Koh-en Yamauchi
3
1Miyazaki Midori Pharms. Inc., 945 Hieda, Akae, Miyazaki-city, 880-0912, Japan
2Animal Resource Center, Graduate School of Agriculture and Life Sciences, The University of Tokyo,
Ago 3145, Kasama, Ibaraki, 319-0206, Japan3Laboratory of Animal Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan
Effects of wood vinegar (WV) on red mites, safety test for chicks, and egg production performance were
examined. In the counting of red mite numbers at cage knots using RGB color range of red mites by the image
analyzer, the red mite were decreased after spraying (P<0.01). In a WV safety test for 8-day-old chicks, one ml of
water, original WV, or WV diluted 500 and 1,000 times were tube-fed twice daily for 6 days. The rate of body weight
gain was decreased in original WV group than that in the control (P<0.05), but it was not different in both WV
dilution groups compared with that in the water-only group. In spraying one ml WV to the hen’s face twice per day
for 8 days, the egg production performances of WV group did not changed so much compared with those of the
control and initial day. These results suggest that the WV did not reduced production performance.
From these results, WV could exterminate red mites, and did not reduce egg production, suggesting that WV is a
useful natural substance to exterminate red mites without harmful effect.
Key words: egg production, extermination, laying hen, mortality, red mite, wood vinegar
J. Poult. Sci., 51: 327-332, 2014
Introduction
Red mites (Dermanyssus gallinae, poultry red mite,
chicken mite) are the most important epidemiological and
economical problem for the poultry industry worldwide.
This red mite is a blood-sucking pest that causes losses in
poultry production because of irritation and anemia. Red
mite-infected hens self-groom and scratch their heads during
the day and night (Kilpinen et al., 2005). Infected hens also
show reduced weight gain (Chauve, 1988; Kilpinen et al.,
2005), egg production, and egg quality (Chauve, 1988;
Cencek, 2003), as well as anemia and death (Kilpinen et al.,
2005). In some case, red mites cause staining of the egg
shell surface (Cencek, 2003). Therefore, poultry farmers
must clean empty houses between flock cycles with 70℃
high-pressure steam to control the red mites. Furthermore, a
variety of chemical medicines have been also used. How-
ever, continued use of chemicals has induced the develop-
ment of drug-resistant red mites. Furthermore, chemical use
results in environmental contamination. Therefore, alterna-
tives are being developed to control red mites and to reduce
environmental pollution through the use of natural resources.
Traps impregnated with neem oil originating from the
Azadirachta indica tree significantly reduce red mite field
populations (Lundh et al., 2005), and pure garlic juice
quickly kills these mites (Maurer et al., 2009). These reports
suggest that plant-derived compounds may be useful to
control red mites.
A wood vinegar (WV) solution was prepared after cooling
smoke during charcoal preparation from the bark of broad-
leaf trees. As WV includes phenols (Matsui et al., 1998),
and inhibits the growth of Salmonella Enteritidis but stimu-
lates the growth of Enterococcus faecium (Watarai and Tana,
2005), we conducted a preliminary test to determine whether
WV could kill red mites, resulting in that the red mites
disappeared from the chicken cages after spraying 500 di-
lution WV.
Therefore, purpose of the present work was to establish
whether the WV can exterminate red mites without lowering
egg production performance by spraying WV on red mites
and chickens.
Materials and Methods
Effects of Spraying WV on Red Mites in Cage Knots
Eight points of cage in a hen laying house knots were
Received: September 17, 2013, Accepted: November 6, 2013
Released Online Advance Publication: December 25, 2013
Correspondence: Y. Matsumoto, Laboratory of Animal Science, Faculty of
Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan.
(E-mail: [email protected])
http:// www.jstage.jst.go.jp / browse / jpsa
doi:10.2141/ jpsa.0130170
Copyright Ⓒ 2014, Japan Poultry Science Association.
randomly selected as observation points (treatment: four
points were sprayed with 1ml of 500 dilution WV, control:
others were not spraying). To analyze the exterminating
effect of WV to red mites, each point was taken a photograph
with a digital camera. The numbers of red mites appeared on
the screen were estimated by the color range of photograph
using the image analyzer (Win Roof, MITANI
CORPORATION, Tokyo, Japan). Color range was meas-
ured using red, green and blue color code (RGB) (range is 0
to 255), and expressed by dot numbers (one dot was 0.02
mm2). In the case of red mite, 82 to 99 color range of red,
47 to 67 color range of green, and 69 to 82 color range of
blue (total 5292 RGB color range) were identified as a color
of red mites, and mean size of red mites was 36 dots (0.72
mm2). After 24 h spraying to treatment points, the eight
points of cage knots were again taken a photograph, and
RGB color ranges were compared between control and
treatment groups.
WV Safety Test on Chicks
A safety test was performed according to the “chick
growth test” on the basis of the safety assessment standard
for the feed and evaluation procedure (the 597th
20 con-
sumption life article) from the Ministry of Agriculture,
Forestry and Fisheries consumption safety chief on May 18,
2008. Twenty-four Sanuki Cochin male chicks (8 days old)
were tube-fed water, original WV, or WV diluted 500 and
1,000 times with water. One ml of each solution was given
twice daily in the morning and evening for 6 days. Starter
mash diet for layers (CP, 21.0%; ME, 3,000 kcal/kg) was fed
ad libitum, and initial and final body weights were measured
to calculate body weight gain rate (body weight gain/initial
body weight). Lethal Dose, 50% (LD50) value was calcu-
lated using the formulas:
(A/6+B/6)÷2=C,
2 g×6 days=12÷C
where,
A=initial body weight of six birds
B=final body weight of live birds
C=mean body weight
Effects of Spraying WV on Egg Production Performance in
Laying Hens
Sixteen hens (Sonia strain, 24-week-old) with similar egg
production performance were selected. They were continu-
ously fed the same diets (CP, 17%; ME, 11.93MJ/kg), and
divided into 2 groups: control and WV spraying groups. One
ml of original WV solution was sprayed on the hen’s face
twice per day for 8 days in WV group (control was only
feeding conventional basal diet without spraying). Feed
intake, hen-day egg production, egg mass weight, egg
weight, feed efficiency, shell-breaking strength, shell
thickness, shell ratio, albumin ratio, yolk ratio, yolk color,
and Haugh units before spraying WV were compared with
those after WV spraying to hens.
Statistical Analysis
The numbers of red mites, body weight gain and egg
production performance were analyzed by one-way analysis
of variance with the Statistical Package for the Social Sci-
ences software version 10.0 for Windows (SPSS, Inc.,
Chicago, IL, USA). In the RGB color range and production
performance, significant differences between groups were
analyzed using independent samples t-test. Differences were
considered significant at P<0.05. In body weight gain, dif-
ference among treatment groups was assessed by Duncan’s
test and were considered significant at P<0.05. The pro-
duction performance and numbers of red mites between
before and after spraying were assessed by a paired samples
t-test in SPSS at P<0.05.
All experiments and collection protocols in the present
study were managed in accordance with the guidelines and
rules for animal experiments, Kagawa University, Japan.
Results
Effects of Spraying WV on Red Mites in Cage Knots
Fig. 1 shows example pictures of cage knots during
experiment. Many red mites appeared on cage surfaces from
the inner part of cage knots, and many red mite eggs were
destroyed at points 1 and 2, but not 3 after spraying the WV.
Table 1 shows the RGB color ranges in cage knots of two
groups were 29±7 and 59±14, respectively (Table 1; P=
0.329). Then, one group showing value of 29±7 was not
sprayed WV (control); the group of 59±14 was sprayed WV
(treatment). The RGB color ranges on the next day was 40±
4 in the control group, but 10±2 in the treatment group,
resulting in that the RGB color ranges of treatment group
decreased than those of the control (P<0.01). When these
values of 29 and 40 were calculated as an increased ratio (%),
it was +37±29 in the control group. This means that the
numbers of red mites showed 37% increase in a day (P<
0.01). In the case of the treatment group, the increased ratio
was −91±5; the numbers of red mites showed 91%
decrease in a day after spraying WV than those before
spraying it (P<0.01).
WV Chick Safety Test
Two birds in the original WV group were dead on days 4
or 6 after starting the safety test. The rate of body weight
gain was significantly lower in the original WV group (P<
0.05) but was not different in the 500 and 1,000 dilution WV
groups compared with that in the water-only group (Table 2).
Although there was no systematic attempt to quantify other
measurement, no differences were observed in behavior,
feather color, or feces among the groups. Calculated LD50 of
the original WV group for chicks was 126,316mg/kg.
Effects of Spraying WV on Egg Production Performance in
Laying Hens
Table 3 shows the egg production performance of the
control (no spraying) and WV group on initial (upper) and
final (lower) days for 8-day-raising. Egg production per-
formance was not different between groups both on initial
and final days. Compared with values of production per-
formance on the initial day, yolk color value was higher on
the final day (P<0.05). In the WV group, the albumin ratio
(P<0.05) was lower, but egg mass weight (P<0.05) and
shell ratio (P<0.01) were higher than those of the initial day.
Journal of Poultry Science, 51 (3)328
Discussion
The current egg layer breeding method has changed from a
floor feeding system to a large-scale cage feeding system
with high bird densities. This change in feeding system has
promoted development of red mites, and stressed birds,
resulting in poor production performance. Red mites live in
the knots of cages, on the egg conveyor belt, or hidden in
cracks and crevices where they lay eggs during the day.
Because the feeding‒oviposition cycle of the red mite repeats
about every third day (Kirkwood, 1963; Desch, 1984), its
life cycle can be completed within 1 week, and large popu-
lations can be rapidly established in poultry houses (Chirico
et al., 2003). Therefore, poultry industry needs an effective
insecticide with sustained but not harmful effect.
A variety of chemical medicines such as organo phos-
phorous compounds (dichlorvos, malathion, metriphonate),
carbamates (propoxur), and synthetic pyrethroids (cyfluthrin
and pyrethrin I and II) were used in poultry house (Höglund
et al., 1995; Chirico and Tauson, 2002; Murano, 2007;
Yamauchi et al.: Exterminating Effect of Wood Vinegar 329
+37±29
Treatment (4)
After 40±4
Before 29±7
Table 1. Effect of spraying wood vinegar (WV) on red, green and blue (RGB) color
range of cage knots
(/cm2)
RGB color range
Control (4)
Values are means±SEM of the number of observed points in parentheses.
−91±5
0 .005
0 .329
p-value
59±14
Increased ratio (%)
10±2
Fig. 1. Red mites on cage knots before and after wood vinegar (WV)
spraying. 1 & 2: WV spraying, 3: no WV spraying.
Murano et al., 2008). However, these reports show that
many chemicals were difficult to exterminate red mites per-
fectly in a short term. The result of spraying to cage knots
revealed 91% of red mites was disappeared by the present
WV solution after 24 h spraying (Table 1). Although there
was no systematic attempt to quantify other measurement,
many red mite eggs were destroyed and their eggs could not
be found at WV spraying points 1 and 2 (Fig. 1). Col-
lectively, these results indicated that WV had strong in-
secticidal effect, but not evasion effect on the red mites in a
short term.
As for the chemical medicines as mentioned above, the
toxicity becomes the problem (Kim et al., 2007). Actually,
when hens with red mites had soaked and their cages were
Journal of Poultry Science, 51 (3)330
Mortality
82 .5±4 .2a
85 .8±4 .4a
56 .3±10 .1b
Table 2. Effect of oral administration of wood vinegar
(WV) solution on body weight gain and mortality in
chicks (mean±SEM, n=6)
1,000 WV
77 .8±2 .9a
Water
Body weight gain rateGroup
Means with different letters are significantly different at p<0.05.
500 WV: wood vinegar diluted with water at 500 times, 1,000 WV:
wood vinegar diluted with water at 1,000 times.
500 WV
0/6 (0%)
0/6 (0%)
2/6 (33%)WV
0/6 (0%)
Yolk color
WV
12 .5±0 .3
Table 3. Effect of spraying wood vinegar (WV) on egg pro-
duction performance in laying hens (mean±SEM, n=4)
75 .1±4 .7
Feed intake
Haugh unit
Items
Means with different superscripts differ from upper and lower significantly
(p<0.05).
Albumin ratio (%)
59 .4±0 .4b
27 .7±0 .4Yolk ratio (%)
29 .6±0 .3
98 .5
12 .0±00
81 .7±1 .2
0 .37±0 .15Shell thickness
0 .41±0 .16(mm)
105 .3
10 .1±0 .3a
Shell ratio (%)
11 .0±0 .4b
(g/hen/day)
62 .2±0 .7a
65 .5±1 .3
0 .65Feed efficiency
0 .62
2 .88±0 .22Shell-breaking
2 .68±0 .58strength (kg/cm2)
Control
75 .0±9 .6Hen-day egg
82 .0±4 .0production (%)
36 .3±3 .8a
Egg mass weight
54 .5±2 .2b
(g/hen/day)
64 .2±1 .8Egg weight
50 .5±2 .4
46 .0±3 .2
78 .5±4 .0
86 .0±4 .0
104 .8
90 .4
2 .68±0 .45
0 .61
0 .69
64 .0±1 .5
62 .4±2 .4
10 .3±0 .1
9 .9±0 .5
0 .39±0 .18
0 .36±0 .21
2 .78±0 .11
13 .0±0 .0(b)
11 .8±0 .3(a)
27 .9±1 .2
27 .2±0 .3
61 .8±1 .1
62 .9±0 .8
─
─
P-value
75 .8±5 .2
80 .8±3 .6
0 .505
0 .580
0 .269
0 .100
0 .554
0 .129
0 .562
0 .872
0 .703
─
─
0 .109
0 .500
0 .177
0 .672
0 .437
0 .310
0 .370
0 .182
0 .391
0 .246
0 .476
sprayed every day for two weeks using the custom method
(soak; 1,000 times diluted pyrethroid insecticide solution,
spraying: a mixed solution of liquid paraffin and pyrethroid
insecticide), we found that the treatment could not control the
red mites while egg production rate gradually decreased to
30% and birds began to die (unpublished data). According to
Poisonous and Deleterious Substances Control Law, a poison
has an LD50 <50mg/kg, and deleterious compound has an
LD50 <300mg/kg. In rats, the LD50 was 5,000mg/kg
(Watanabe et al., 2008). In this study, the present higher
value of LD50 of 126,316mg/kg than that of rat was safe for
chicks, suggesting that diluted WV has no harmful effect on
chickens. Moreover, WV spraying the hen’s face with WV
may not have induced stress because their egg production
performances did not changed (Table 3). Therefore, it is also
possible that WV can improve poultry production. However,
the albumin ratio was lower than those of the initial day.
Further work is needed to investigate the effect of WV
spraying on egg quality.
Although it is considered that they suck blood from hens at
night and leave before daybreak (Arends, 1991), they para-
sitize and propagate on the chickens day and night without
leaving the chickens before daybreak (Nakamae et al., 1997).
Thus, daytime spraying could be useful for anthelmintic of
red mites. In fact, we observed in spraying a 200-ml solution
of 500 times diluted WV with tap water to red mites ac-
cumulating on hen’s feathers around the cloaca (Left in Fig.
2), the WV could exterminate these red mites on day 4 after
spraying (Right in Fig. 2).
Another problem for chemical medicines is the develop-
ment of drug resistance. Repeated long-term control of red
mites with the chemicals induces the development of heri-
table resistance in the mites (Zeman, 1987). It is likely that
the WV might not induce such a resistance in the mites,
because the WV is natural substance. In the report using
natural substances, cardboard traps containing 20% neem oil
from the tree Azadirachta indica induced a 92% reduction of
red mites after placing traps at the mites’ aggregation sites
for four weeks (Lundh et al., 2005). These reports imply
that natural substance such plant can also control the red
mites without drug resistance.
In conclusion, WV could exterminate red mites and spray-
ing WV on hens did not reduce egg production, suggesting
that WV is a useful natural substance to control red mites
without lowering egg production performance.
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