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Toxic Impacts of Sub-Chronic Inhalation of Mosquito Coil Smoke in RabbitsGhani Nadia and Shahbaz Anum Department of Environmental Sciences, Lahore College for Women University, Jail Road, Lahore, PAKISTANAvailable online at: 
www.isca.in, www.isca.me
Received 24th April 2014, revised 1st May 2014, accepted 19th May 2014 AbstractVarious Asian countries including china, Thailand, India, Malaysia Korea and Japan commonly used mosquito coils as mosquito repellent. Mosquito coils are slow-burning product which release smoke containing one or more insecticides; it burns for several hours in confine places in order to provide protection to humans against malaria, which spread due to mosquitoes. Present study was intended to examine the toxic impacts of inhaling mosquito coil smoke on male rabbits. Mosquito coils are widely used to kill mosquitoes. A total of forty rabbits were used in this study. These rabbits were divided into two major groups i.e., Control Group and Experimental Group. Experimental group was further divided into 3 sub-groups i.e., E1, E2 and E3.Each group contain 10 rabbits as shown in figure 2.1. Control Group was not exposed to mosquito coil smoke while Experimental groups were exposed to mosquito coil smoke for 3 hours/day. Exposure time remain the same but exposure days vary with each Experimental group i.e., E1 was exposed for 5 days, E2 was exposed for 10 days and E3 was exposed for 15 days. After the completion of exposure day’s blood was collected from the ear pinna of rabbits for the analysis of Red Blood Cells, White Blood Cells and Platelets. Blood samples were analyzed by using Hematology Analyzer. Effect of mosquito coil smoke on weight of rabbits was also observed and morphological changes due to mosquito coil smoke was also evaluated. Results were analyzed by comparing the data of Experimental groups with Control group. According to results a significant decrease was observed in weight of Group E3 rabbits. Results also demonstrated that with the increase of exposure day’s number of White Blood Cells and Platelets were also increased while the numbers of Red Blood Cells become decreased. Keywords: Red blood cell, white blood cell, platelets, complete blood count. IntroductionThe urban population is expected to increase more than 70% in developed countries resulting in the increased usage of urban pests. Recently, several new mosquito control devices are available in markets in order to reduce or eliminate the number of mosquitoes by effectively trapping or preventing them from residential properties. Mosquito coils are slow-burning product which release smoke containing one or more insecticides; it burns for several hours in confine places in order to provide protection to humans against malaria, which spread due to mosquitoes. Mosquito coils are widely used as a mosquito repellent in various Asian countries including china, Thailand, India, Malaysia Korea and Japan. Burning one mosquito coil would release same amount of PM2.5 mass which is equivalent to burning 75-137 cigarettes. A World health organization (WHO) report estimated the world wide annual consumption of mosquito coils to be approximately 29 billion pieces. Most of mosquito coils consist of an active ingredient known as pyrethroids, consist of about 0.3-0.4% of the coil mass. Other ingredients of mosquito coil includes various binders, fillers, dyes, d-allethrin along with wood powder, coconut shell powder starch and additives capable of smoldering. Many mosquito coils also contain a chemical term S2 (octa-chlorodipropyl ether) during burning of coil S2 degraded into another chemical known as bis-chloromethyl ether (BCME), it is a potential cancer causing agent. Epidemiological studies have shown that mosquito coil smoke can stimulate asthma and persistence wheeze in children when they are exposed to it for a long period of time. There are also some irritants which are released in the coil smoke cause sneezing in rats such as aldehydes and sulphates. Animal studies with long term exposure to mosquito coil smoke showed abnormal growth of skin cells, per weight loss and lung damage10. Hematological studies on rats exposed to mosquito repellents carried out by Ayorinde et al. (2012), in this study rats were divided into two groups of 6 males and 6 females and they were exposed to mosquito repellents per day for four weeks. The results showed increase in white blood cell (WBC) counts in the females and males exposed for two weeks. Platelets (PLT) counts in female rats (after two weeks) also increased when compared with the control group11. Material and Methods Experimental Rabbits: The present study was undertaken to determine the toxic effects of mosquito coil smoke on 
International Research Journal of Environment Sciences______________________________________________ ISSN 2319–1414 Vol. 3(5), 86-91, May (2014)     Int. Res. J. Environment Sci. International Science Congress Association             
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mammals, for this purpose male rabbits were used. Total of 40 adult male rabbits free from any apparent clinical ailment were purchased from the local market. Rabbits were kept in wire cages in Botanical garden of Lahore College for Women University and they were divided into two main groups:1)Control group (C), and2)Experimental group. Fresh grass and green fodder and drinking water were provided, around the clock.  Mosquito Coil: The commonly used brand of mosquito coil (Mortein Coil) was purchased from local Market. Mosquito coil used measured 12 cm diameter and 15.0 g weight.  Experimental Design: Control group comprised of 10 rabbits and they were exposed to normal room (125m) fresh air. While Experimental group comprised of total 30 rabbits and they were further divided into three groups of E1, E2 and E3. Each group consisted of 10 rabbits E1, E2, and E3. E1 group was exposed to mosquito coil smoke for 5 days/3 hrs, E2 group were exposed for 10 days/3 hrs and E3 group were exposed to coil for 15 days/3 hrs respectively. They were marked with different colors permanent markers (blue, red and green) for individual identity.1ml blood was collected from the ear pinna of rabbits before and after the exposure. Parameters: The parameters to be studied include: i. Morphological and behavioral changes, ii. Body weight, iii. Complete blood count (CBC). Instruments: i.Hematology Analyzer (Model:Sysmex KX-21), ii. Weighing balance Apparatus: For Tagging: i. Permanent markers. For Exposure: i. Mortein mosquito coil, ii. Match box. For Blood Collection: i. Syringes, ii. Cotton, iii. EDTA blood collection glass tubes, iv. 90% Ethanol. Body Weight Analysis: Electrical weighing balance of Model ALPHALA, range: 35 kg was used for calculation of subsequent changes in weight due to mosquito coil smoke. Initial, continuous and final body weights of male rabbits after exposure period of mosquito coil smoke were documented according to their group. Each value was counted twice and their mean value was taken for the comparison of weight change between control group and experimental groups. Exposure to Mosquito Coil Smoke: Mosquito Coil smoke was got inhaled by rabbits according to their groups. Coil was placed 6 inches away from rabbits and cages were covered from sides so that proper circulation of smoke occur. About 3 hours exposure had been given to rabbits daily according to their exposure days as shown in figure 1. Control group remained in fresh air. Blood Sampling: Blood was collected from both experimental rabbits as well as control ones. Rabbit’s ear pinna veins (intravenous blood collection) were chosen for blood collection. The auricular vein is a common venipuncture site in rabbits. It can easily be seen on the dorsal surface of ear in the center between the two marginal ear veins. Before blood collection withdrawal site was cleaned with 90% Ethanol as disinfectant. The ear had to warm or tapped with fingers in order to dilate the vessels. Sterile 23G (gauge) needle with BD 5 ml syringes was used for blood collection and also to limit tissue trauma and discomfort.1 ml blood was drowned at each collection from every rabbit. The marginal ear veins were used to collect up to 1 millimeter of blood samples from the tip of the ear, away from the base of the ear. Applying finger pressure on cotton placed at the blood sampling site for approximately 2 min in order to stop the blood flow before rabbits returned to its cages.  According to rabbit exposure and defined days blood samples were collected as taken after 5 days from E1 group rabbits, after 10 days from E2 group rabbits and after 15 days from E3 group rabbits correspondingly. Collected Blood was poured in EDTA blood collection glass tube and slightly shakes to prevent clotting formation. Hematological Analysis: The blood samples of experimental rabbits and control group rabbits were analyzed to test the effect of mosquito coil smoke on complete blood counts (CBC) of rabbits. For this purpose hematology analyzer (Model: Sysmex KX-21) as shown in figure; was used to analyze the following: White blood cell (WBC), Red blood cell (RBC), Hemoglobin (HGB), Platelets (PLT), Lymphocytes(LYM), Neutrophils (NEUT), Red cell distribution width (RDW), Platelet distribution width (PDW), Mean platelet volume (MPV), and Ratio of large platelets (P-LCR). Statistical Analysis: Data of study was investigated through comparing all the experimental data with control. Data were expressed as mean ± standard error of the mean. Statistical significant was performed using the analysis of variance (ANOVA).
Results and Discussion
Observed morphological toxicity symptoms in present study 
were include Wheezing, Sleepiness, Nausea, Slight salivation, 
Slight eye irritation, Lacrimation, Heavy fits, Productive cough 
and Scratching of nose. While weakness, laziness and loss of 
appetite were observed in each rabbis of all Experimental 
groups. 
 
Pyrethroid is considered as an important ingredient of mosquito 
coil. Exposure to very large amounts of pyrethroids leads to 
dizziness, headache, and nausea that might last for several 
hours. Larger amounts could cause muscle twitching, reduced 
energy, and changes in awareness. Even larger amounts could 
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cause convulsions and loss of consciousness that could last for 
several days12 
 
According to the research conducted by Baron ,this study 
revealed that Cyanide, which is released through coil smoke, 
reduces the oxygen carrying capacity of erythrocytes, which 
leads  to reduce metabolism and energy output which indicates 
the weakness of body13. 
An infection can stimulate the body to produce higher number of bacterial infection. An increased number of lymphocytes may be produced with the viral infection. In certain states of diseases such as leukemia, abnormal white blood cells multiply rapidly ultimately leading to increase the number of WBCs count. Present study also investigated the increase in white blood cells count in all experimental groups after mosquito coil smoke inhalation. 
Red Blood Cells are also known as Erythrocytes. They are produced in bone marrow and released into the blood stream as they become mature. If insufficient amount of RBCs are present then in that condition person suffer from anemia, fatigue and weakness14
 In present study the amount of RBCs count was decreased. Control group did not show any change in RBCs count while experimental groups showed slight decrease in RBCs count. Figure-1 Flow sheet Diagram showing Mosquito Coil Smoke Exposure Pattern and Group Division 
Table-1 
Effect of Mosquito Coil Smoke on Weight (kg) Experimental Groups Control Group(C) ANOVA(Group wise Comparison) 
E1 Group  (5 days exposure) 1.1530±0.0471 1.4370±0.0710 ** 
E2 (10 days exposure) 1.0630±0.0641 *** 
E3 (15 days exposure) 1.0470±0.0714 *** 
ANOVA (Intergroup Comparison) *** 
Values given are Mean ± SE of results obtain from forty rabbits. X=Insignificant difference of mean value, =Significant difference (P0.05), **=More Significant difference (P0.01), ***=Highly Significant difference (P0.001)11. Table-2 Effect of Mosquito Coil Smoke on White Blood Cells (10/µL) Experimental Groups Control Group(C) ANOVA(Group wise Comparison) 
E1 Group (5 days exposure) 4.760±0.428 5.170±0.705 X 
E2 (10 days exposure) 6.240±0.554 X 
E3 (15 days exposure) 8.000±0.905 * 
ANOVA (Intergroup Comparison) ** 
Values given are Mean ± SE of results obtain from forty rabbits. X=Insignificant difference of mean value, =Significant difference (P0.05), **=More Significant difference (P0.01), ***=Highly Significant difference (P0.001)11 
MOSQUITO COIL SMOKE 
Experimental Group (Exposed to mosquito coil smoke) 
Control Group C 
Group E3 
Group E2 
Group E1 
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Vol. 3(5), 86-91, May (2014)
 
 
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If the numbers of p
latelets count become greater than 
600×10
/µL then myeloproliferative disorders of the stem cells 
occurred in the bone marrow15
. In present study significant 
increase was observed in platelets count of e
xperimental grou
as compared with control group. 
Conclusion 
This study was conducted in order to determine the toxic 
impacts of mosquito coil smoke on b
lood count parameters 
(WBCs, RBCs and PLTs) in male rabbits. Results of study 
showed that mosquito coil smoke induced morphological 
 
Effect of Mosquito Coil Smoke on 
Experimental Groups 
E1 Group (5 days exposure) 
4.9950±0.0819
E2 (10 days exposure) 
E3 (15 days exposure) 
ANOVA (Intergroup Comparison) 
Values given are Mean ± SE of results obtain from forty rabbits
(P0.05), **
=More Significant difference (P0.01)
 
Effect of Mosquito Coil Smoke on Platelets 
Experimental Groups 
E1 Group (5 days exposure) 
E2 (10 days exposure) 
E3 (15 days exposure) 
ANOVA (Intergroup Comparison) 
Values given are Mean ± SE of results obtain from forty rabbits
(P0.05), **
=More Significant difference (P0.01)
 
Histogram showing the comparison of (Mean± SE) values of Weight(kg) of Control and Experimental Groups
0.20.40.60.81.21.41.6E1Weight (kg), (Mean   SE)
Control and Experimental Groups
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latelets count become greater than 
/µL then myeloproliferative disorders of the stem cells 
. In present study significant 
xperimental grou
ps 
This study was conducted in order to determine the toxic 
lood count parameters 
(WBCs, RBCs and PLTs) in male rabbits. Results of study 
showed that mosquito coil smoke induced morphological 
symptoms in e
xperimental groups which include 
Wheezing, Sneezing, Slight eye irritation, 
salivation, heavy fits, c
onvulsions, 
of nose and weakness. Mosquito coil smoke also decreased the 
weights of Experimental Group rabbits. This study also showed 
that as the number of exposure days increased but the exposure 
time remai
n the same then the amount 
PLTs (10
/µL) count were increased while amount of RBCs
(10/µL) 
count become decreased as compared with Control 
Group which exposed to fresh air. Table-3 
Effect of Mosquito Coil Smoke on 
Red Blood Cells (10/µL) Control Group(C) 
4.9950±0.0819
5.668±0.157
5.250±0.101
4.757±0.175
*** 
Values given are Mean ± SE of results obtain from forty rabbits
. X
Insignificant difference of mean value
=More Significant difference (P0.01)
, ***=Highly Significant difference (P0.001)11. Table-4 
Effect of Mosquito Coil Smoke on Platelets 
(10/µL) Control Group(C) 
659.5±40.1521.3±60.4
769.2±53.5
917.0±86.5
*** 
Values given are Mean ± SE of results obtain from forty rabbits
, X
Insignificant difference of mean value
=More Significant difference (P0.01)
, ***=Highly Significant difference (P0.001)11 Figure 1 
Histogram showing the comparison of (Mean± SE) values of Weight(kg) of Control and Experimental Groups
E2E3
Control and Experimental Groups
Control
Experimental Groups
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xperimental groups which include 
laziness, 
Wheezing, Sneezing, Slight eye irritation, 
lacrimation, heavy 
onvulsions, 
loss of appetite, Scratching 
of nose and weakness. Mosquito coil smoke also decreased the 
weights of Experimental Group rabbits. This study also showed 
that as the number of exposure days increased but the exposure 
n the same then the amount 
of WBCs (10/µL) and 
/µL) count were increased while amount of RBCs
 
count become decreased as compared with Control 
ANOVA(Group wise Comparison) 
** 
X 
** 
 
Insignificant difference of mean value
, =Significant difference ANOVA(Group wise Comparison) 
X 
** 
** 
 
Insignificant difference of mean value
, =Significant difference 
Histogram showing the comparison of (Mean± SE) values of Weight(kg) of Control and Experimental Groups
 
Experimental Groups
International Research Journal of 
Environment 
Vol. 3(5), 86-91, May (2014)
 
 
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After Mosquito coil smoke exposure significant decrease was 
observed in body weight of Experimental Groups as compare to 
Control group. 
Inter group comparison showed highly 
significant difference while in group wise comparison Group E2 
and E3 showed highly significant differences in weights as 
compare to Control Group (figure-1). 
Mosquito coil smoke exposure induced alterat
count of Experimental groups as compare to Control. Decrease 
was observed in Group E1 while Group E2 and E3 showed 
increase in WBCs count. Inter group comparison showed more 
significant differences in WBCs count .On other hand in group 
wise com
parison Group E3 exhibited the significant difference 
 
Bar graph showing the comparison of White Blood Cell Counts 
Bar graph showing the comparison of Red Blood Cell Counts of Control and Experimental Groups
ControlWBCs Count (103 /µL)
4.24.44.64.85.25.45.65.8ControlRBCs Count (106 /µL)
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After Mosquito coil smoke exposure significant decrease was 
observed in body weight of Experimental Groups as compare to 
Inter group comparison showed highly 
significant difference while in group wise comparison Group E2 
and E3 showed highly significant differences in weights as 
Mosquito coil smoke exposure induced alterat
ion in WBCs 
count of Experimental groups as compare to Control. Decrease 
was observed in Group E1 while Group E2 and E3 showed 
increase in WBCs count. Inter group comparison showed more 
significant differences in WBCs count .On other hand in group 
parison Group E3 exhibited the significant difference 
as compare to Control while Group E1 and Group E2 did not 
show statistically significant differences in WBCs count
2). 
Mosquito coil smoke exposure caused reduction in RBCs count 
of Experimenta
l groups as compare to Control. Highest decrease 
was observed in Group E3 which was exposed for 15
group comparison showed highly significant differences in 
RBCs count .While in group wise comparison Group E1 and E3 
exhibited the more 
significant difference as compare to Control 
but Group E2 did not show statistically significant differences in 
RBCs count compared with Control Group
Figure 2  
Bar graph showing the comparison of White Blood Cell Counts 
of Control and Experimental Groups
Figure 3 
Bar graph showing the comparison of Red Blood Cell Counts of Control and Experimental Groups
E1E2E3Control and Experimental Groups
E1E2E3Control and Experimental Groups
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as compare to Control while Group E1 and Group E2 did not 
show statistically significant differences in WBCs count
 (figure-
Mosquito coil smoke exposure caused reduction in RBCs count 
l groups as compare to Control. Highest decrease 
was observed in Group E3 which was exposed for 15
 days. Inter 
group comparison showed highly significant differences in 
RBCs count .While in group wise comparison Group E1 and E3 
significant difference as compare to Control 
but Group E2 did not show statistically significant differences in 
RBCs count compared with Control Group
 (figure-3). 
of Control and Experimental Groups
  
Bar graph showing the comparison of Red Blood Cell Counts of Control and Experimental Groups
 
Control
E1
E2
E3
Control
E1
E2
E3
International Research Journal of 
Environment 
Vol. 3(5), 86-91, May (2014)
 
 
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Bar graph showing the comparison of Platelet Counts of Control and Experimental Groups
Explanation: 
Mosquito coil smoke exposure increased the 
Platelets count of Experimental groups as compare to Control. 
Highest increase was observed in Group E3 which was exposed 
for 15 days to mosquito coil smoke. Inter group comparison 
showed highly 
significant differences in Platelets count .While 
in group wise comparison Group E2 and E3 exhibited the more 
significant difference as compare to Control but Group E1 did 
not show statistically significant differences in Platelets count 
compared with Control Group (figure-4). References 1.Watson C., 
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the 1st
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Urban Environment. 
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Figure 4 
Bar graph showing the comparison of Platelet Counts of Control and Experimental Groups
Mosquito coil smoke exposure increased the 
Platelets count of Experimental groups as compare to Control. 
Highest increase was observed in Group E3 which was exposed 
for 15 days to mosquito coil smoke. Inter group comparison 
significant differences in Platelets count .While 
in group wise comparison Group E2 and E3 exhibited the more 
significant difference as compare to Control but Group E1 did 
not show statistically significant differences in Platelets count 
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E1E2E3Control and Experimental Groups
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Bar graph showing the comparison of Platelet Counts of Control and Experimental Groups
 
Researchers concerned over 
effects of 
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Control
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