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International Journal of Advanced Technology & Science Research
Volume 01 Issue 01 January 2019
www.ijatsr.org Copyright © IJATSR 2019, All Rights Reserved Page 46
THE EFFECT OF TEMPERATURE AND COUMAPHOS
CONCENTRATION ON THE MORTALITY RATE OF
BOOPHILUS MICROPLUS LARVAE
Hamong Suharsono1 and Kadek Karang Agustina
2
1Laboratory of Biochemistry Faculty of Veterinary Medicine Udayana University Denpasar Bali, Indonesia.
2Department of Veterinary Public Health Faculty of Veterinary Medicine Udayana University Denpasar Bali, Indonesia
ABSTRACT
This study aims to determine the effect of temperature and various coumaphos concentrations
on the survival of Boophilusmicroplus larvae in either the highlands or lowlands on the island
of Bali Indonesia. The sample used was the larva stage of B.microplus obtained by breeding
female B.microplus ticks until the eggs hatched. Coumaphoswas prepared in sixs different
concentration. All treatments separate in 2 temperature levels ie. 18o to <25
oC and 25
oto
31oC.The results showed that in the temperature of 18
o to <25
oC, the concentration of
coumaphos of 5x10-1
to 5x10-2
showed the effective treatment for killing B. micropluslarvae
up to 100%. Better results obtained in the temperature of 25oto 31
oC which the effective
treatment of coumaphos was in the concentration of 5x10-1
to 5x10-3
. To conclude, the higher
temperature increases the effectivenes of coumaphos to kill B. micropluslarvae.
Keywords: asuntol, Boophilusmicroplus, coumaphos, concentration, temperature.
INTRODUCTION
Indonesia as a tropical country with high relative humidity [1], that there is a suitable place
for the life of various ectoparasites [2]. Among the many ectoparasites that attack cattle,
goats, sheep and horses, B. microplus is one of the most important and often found [3]. These
ticks can act as vectors for Babesiosis, Anaplasmosis and Theileriosis [4]. Dennig (1975) [5],
proved that experimentally B. microplus can transmit Jembrana disease in Bali cattle. Apart
from being able to transmit the disease, these ticks are blood sucking ectoparasites for their
survival [6]. With continuous blood loss anemia can occur, which can eventually weaken the
physical condition of the happy parent [7]. In addition, tick bites can cause skin damage
which can be a predisposing factor for other secondary infections [8].
International Journal of Advanced Technology & Science Research
Volume 01 Issue 01 January 2019
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B. microplus is one of the ectoparasites, which is very disturbing to livestock both directly
and indirectly [9], this ticks larvae also known as zoonotic agent [8]. Control of ticks can be
done through management of pastures or using chemicals [10]. In Indonesia drugs that are
often used for tick control are Etion, Trichlorofon and coumaphos. Coumaphos is one type of
neurotoxic organophosphates insecticide that is widely used [11]. Andrew et al. (1976) [12],
stated that insecticides and acarisides must be efficacious and safe for livestock.
Based on the fact that the tick infestation in livestock is still high and the use of coumaphos
as insecticides in its control efforts is increasingly used, it is necessary to do research on the
effect of temperature and coumaphos concentration on the survival of B. microplus larvae.
MATERIALS AND METHODS
Samples
This study used B. microplus larvae obtained by breeding of female B. microplus until the
eggs hatch and the larvae were collected. We used a total of 1.200 B. microplus larvae as a
sample in the study.
Preparation of coumaphos solution
Using the Sartorius scale, weighing 1 gr Asuntol® 50 WP powder (contain 50% of
cumaphos) for making the various concentration solution. Coumaphos solution of 5x10-1
%,
made by dissolving 1 gram of Asuntol® 50 WP by distilled water until 1 lt of volume. 5x10-
2% coumaphos solution, made by adding 90 ml of distilled water to 10 ml of 5x10
-1%
coumaphos solution. 5x10-3
% coumaphos solution, made by adding 90 ml of distilled water
to 10 ml of 5x10-2
% coumaphos solution. 5x10-4
% coumaphos solution, made by adding 90
ml of distilled water to 10 ml of 5x10-3
% coumaphos solution. and 5x10-5
% coumaphos
solution, made by adding 90 ml of distilled water to 10 ml of 5x10-4
% coumaphos solution.
As a negative control, sterile distilled water was used (0.00% of cumaphos).
Research design
The study was conducted using a completely randomized factorial 2X6 pattern [13]. In this
term, two levels of environmental temperature factors, namely temperatures of 18o to <25
oC
and 25o
to 31oC (which are adjusted for the temperature in the highlands and lows of the
island of Bali) and six levels of coumaphos concentration factors (5x10-1; 5x10-2; 5x10- 3;
5x10-4; 5x10-5 and 0.00%), as a combination of treatments. Repetition were done four times
for each combination of treatments. And each combination of treatment was estimated to be
between 10-60 larvae as the study sample (Tab 1).
International Journal of Advanced Technology & Science Research
Volume 01 Issue 01 January 2019
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Table 1. The number of B. microplus larvae used in the study
Temperature
Coumaphos concentration (%) Number of larvae
Total
I II III IV
18o to <25
oC
5x10-1
28 29 25 29 111
5x10-2
20 26 32 23 101
5x10-3
36 19 18 17 90
5x10-4
27 23 34 24 85
5x10-5
18 41 29 31 119
0,00 21 34 27 26 108
25o to 31
oC
5x10-1
21 59 16 21 117
5x10-2
19 19 13 22 83
5x10-3
18 11 24 26 79
5x10-4
16 35 1 19 88
5x10-5
15 19 24 30 88
0.00 31 26 25 26 108
Total 270 341 295 294 1200
Treatments
Each prepared larvae group (Tab 1) is wrapped into gauze and dipped for 1.5 minutes at each
concentration of solution. It was dried and then left overnight according to the different
temperature [14]. Larvae was observed under stereo microscope to see the effect of
treatments. The benchmark of this study was the number of dead larvae that are determined
by looking at the movements of the larvae. If the larva is still able to move one or two feet,
but when touched the larvae remain silent, then this was also expressed as the death of larvae
due to the treatments [15].
Data analysis
International Journal of Advanced Technology & Science Research
Volume 01 Issue 01 January 2019
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Data of the percentage of death of B. microplus larvae were analyzed by Variety
Investigation Analysis. Before being analyzed, the data in the percentage were transformed
first with the Arc Sin transformation method [16-17]. If the results were significantly
different, the analysis wass continued with Duncan's Multiple Distance Test [17-18].
RESULTS
The obtained data of this research are present in table 2 to 4 below.
Table 2. The percentage of death of B. microplus larvae after treatments.
Temperature Coumaphos concentration
(%)
Death of larvae (%) Average
(%) I II III IV
18o to
<25oC
5x10-1
100.00 100.00 100.00 100.00 100.00
5x10-2
100.00 100.00 96.87 100.00 99.22
5x10-3
61.11 36.84 33.33 29.41 40.17
5x10-4
11.11 26.09 17.65 29.16 21.00
5x10-5
11.11 12.20 13.79 3.23 10.83
0.00 14.28 2.94 7.40 15.38 10.00
25o to 31
oC
5x10-1
100.00 100.00 100.00 100.00 100.00
5x10-2
100.00 100.00 100.00 100.00 100.00
5x10-3
100.00 100.00 100.00 100.00 100.00
5x10-4
87.50 65.71 66.67 68.42 72.08
5x10-5
33.33 42.50 25.00 23.33 30.94
0,00 3.22 2.85 16.00 23.30 11.34
Average (%) 60.14 57.43 56.76 57.69 57.97
International Journal of Advanced Technology & Science Research
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Table 3. Duncan's Multiple Distance Test on the effect of combination treatment on the death
rate of B. microplus larvae (Arc Sin Transformation)
Treatment Average of death
larvae (%)
ArcSin
Transformation
Value
Sig. (0.05) Group
25o to 31
oC, 5x10
-1 100.00 90.00 A
I
25o to 31
oC, 5x10
-2 100.00 90.00 A
25o to 31
oC, 5x10
-3 100.00 90.00 A
18o to <25
oC, 5x10
-1 100.00 90.00 A
18o to <25
oC, 5x10
-2 99.22 87.22 A
25o to 31oC, 5x10-4 72.08 58.50 B II
18o to <25
oC, 5x10
-3 40.17 39.19 C
III 25
o to 31
oC, 5x10
-5 30.94 33.64 Cd
18o to <25
oC, 5x10
-4 21.00 26.94 De
IV
25o to 31
oC, 0,00 11.34 19.79 Ef
18o to <25
oC, 5x10
-5 10.83 18.01 F
18o to <25
oC, 0,00 10.00 17.73 F
Description: The same letter towards the column shows no difference.
Table 4. Duncan's Multiple Distance Test on the Effect of Coumaphos Concentration on the
Death Rate of B. microplus Larva (Arc Sin Transformation)
Coumaphos
concentration (%)
Average of death
larvae (%)
Arc Sin Transformation
Value
Significance
5% 1%
5x10-1
100,00 90,00 a a
5x10-2
99,60 88,61 a a
5x10-3
70,09 64,06 b b
International Journal of Advanced Technology & Science Research
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5x10-4
60,50 42,72 c c
5x10-5
20,84 25,83 d d
5x10-6
10,67 18,76 E d
Description: The same letter towards the column shows no significant difference
DISCUSSION
From the research on the effect of temperature and coumaphos concentration on the survival
of B. microplus larvae, the percentage of died larvae after treatment is shown in Table 2. It
appears that the average of the mortality rate of B. microplus larvae in all treatments was
57.97%. In the temperature of 18o to <25
oC, the concentration of coumaphos of 5x10
-1 to
5x10-2
showed the efective treatment for killing B. microplus larvae up to 100%. Better
results showed in temperature of 25o to 31
oC which the efective treatment of coumaphos were
the concentration of 5x10-1
to 5x10-3
.
From Table 3, we found that there were 5 groups of effectiveness on the combination of
temperature and coumaphos concentration on the mortality rate of B. microplus larvae. Group
I is a combination of treatments with the best result to kill B. microplus larvae. In the group
II, the low concentration of coumaphos (5x10-5
) in the temperature of 25o to 31
oC still give a
good results in killing B. microplus larvae (>70%). Group III and IV showed ineffective
treatment. This findings indicates that in the lower temperature area needs higher
concentration of coumaphos to kill B. microplus larvae.
From the results, it appears that the temperature has a significant effect on the mortality rate
of B. microplus larvae (Tab 3). The mortality of B. microplus larvae treated with various
concentrations of coumaphos at temperatures of 25o to 31
oC resulted in higher mortality of
larvae. This is closely related to the working power of coumaphos which includes
organophosphate insecticides. The toxicity increases with the increase of temperature caused
by the absorption of body cells which increases at a temperature of 25o to 31
oC compared to
temperatures of 18o to <25oC [19-20]. These results are in line with the results of some
previous studies, which states that the workability of insecticides at higher temperatures will
be more effective [21-22].
Table 4 shows that the smaller the concentration of coumaphos, the smaller the mortality rate
of B. microplus larvae and the other way around. The results of this tests confirmed that
coumaphos concentrations for efective treatment of B. microplus larvae were at least 5x10-2
%
with a mortality rate of 99.60%. This data is in accordance to the previous research that found
the use of coumaphos at the dipping vat method reduced tick numbers and reproduction more
effectively than spray-dip or power spray [23].
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Some reports stated that the B. microplus have been resistance to acaricides included
coumaphos [24-26]. B. microplus in cattle in Mexico have been reported be resistance to
coumaphos of 48.3% [24]. The results suggest that an enhanced cytochrome P450
monooxygenase (cytP450)-mediated detoxiÞcation mechanism may exist in the resistant
strains, in addition to the cytP450-mediated metabolic pathway that activates coumaphos.
Triphenyl phosphate (TPP) synergized toxicity of acaricides in susceptible and resistant
strains, and there was no correlation between TPP synergism ratios and the LC50 estimates
for either acaricide. Esterases may not play a major role in resistance to coumaphos.
Resistance to coumaphos in the Mexican strains of B. microplus was likely to be conferred by
both acytP450-mediated detoxication mechanism [25].
CONCLUSION
From the results of the study the effect of temperature and coumaphos concentration on the
survival of B. microplus larvae, it can be concluded that the ambient temperature has a
significant effect on the mortality rate of B. microplus larvae. In this case, the mortality rate
of B. microplus at a temperature of 25 to 31oC was significantly higher than the 18 to <25
oC
at the similar concentration of coumaphos.
COMPETING INTERESTS
All authors declare that there is no conflict of interest related to this article
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