TMJ Vol. 01 No.

02 87-95

Transovarial Transmission Index of Dengue Virus on Aedes

aegypti and Aedes albopictus Mosquitoes in Malalayang
District in Manado, North Sulawesi, Indonesia
Angle Maria Hesti Sorisi1, Sitti Rahmah Umniyati2 , Tri Baskoro Tunggul Satoto2

Postgraduate Program of Basic Medical Science and Biomedical Program, Faculty of Medicine, Universitas
Gadjah Mada, 2Departement of Parasitology, Faculty of Medicine, Universitas GadjahMada.

*Corresponding author: hestisorisi@yahoo.com

ABSTRACT

Introduction: Dengue Hemorrhagic Fever (DHF) is an infectious vector-borne disease caused by Aedes
sp mosquitoes still cause serious health problem in Indonesia. Based on Manado Health Office Report,
Malalayang was identified as dengue-endemic areas. In 2010, number of DHF cases in Malalayang is 211
cases with Incidence Rate (IR) 328 per 100,000 populations. Dengue viruses (DENV) survive in nature by
two mechanisms; by horizontal transmission through infected vertebrates and mosquitoes, and by vertical
(transovarial) transmission in the mosquitoes. Transovarial transmission is assumed as an important aspect
in the maintanance of DENV during inter epidemic, but this problem has not been studied in Malalayang
District, Manado. An effort to prevent and control DHF requires knowledge of an Aedes sp Dengue virus
transovarial infection.
Objectives: To prove the existence of Dengue virus transmission in Ae. aegypti and Ae. albopictus
mosquitoes and its relationship with the incidence of DHF in Malalayang District in Manado, North Sulawesi,
Indonesia.
Methods: The method of this research was an observational analytic study with cross-sectional design.
Study samples were unbloodfed Aedes aegypti and Aedes albopictus mosquitoes on the F1 generation
from ovitrap placed in five selected villages based on the number of cases in the District Malalayang. The
secondary data of DHF patients from Malalayang district was obtained from Health Office Manado and
the Community Health Center in 2010. The presence of dengue antigen in head squashes preparation
were detected using monoclonal antibody against dengue (DSSE10) based on immunohistochemical
streptavidin biotin peroxidase complex (ISBPC) technique to confirm the presence of transovarial
transmission of dengue virus both in Ae. Aegypti and Ae. Albopictus, and to obtain the data of transovarial
transmission index. Fishers Exact test and Pearson correlation are used to analyze those data.
Results: Transovarial transmission of Dengue virus in Aedes sp was found from 5 villages in Malalayang
district with Transovarial Transmission Index (TTI) ranges 6.1%-17.1%. Statistic test showed significant
differences in positive rate (p-value=0.00<0.05) on Ae. aegypti higher than Ae. albopictus. It is also known
that there is no statistically significant correlation (p-value=0.528>0.05) between the Aedes sp. Dengue
virus TTI and DHF IR in Malalayang district.
Conclusion: This study demonstrates the existence of Dengue virus transovarial transmission in Aedes sp
in Malalayang district. Ae. aegyptis TTI is higher than that of Ae. Albopictus, and no significant correlation
between TTI and DHF IR in Malalayang district.
Keywords: DHF, transovarial transmission, Ae. aegypti, Ae. albopictus

INTISARI

Pendahuluan: Demam Berdarah Dengue (DBD) adalah penyakit infeksi yang ditularkan vektor yang
disebabkan oleh nyamuk Aedes sp dan merupakan masalah kesehatan yang serius di Indonesia.
Berdasarkan Laporan Dinas Kesehatan Manado, Malalayang diidentifikasi sebagai daerah endemis demam
berdarah. Pada tahun 2010, terdapat 211 jumlah kasus DBD di Malalyang, dengan IR virus Dengue 328 per

87

Angle M H Sorisi et. al, Transovarial Transmission Index of Dengue Virus on Aedes aegypti and
Aedes albopictus Mosquitoes in Malalayang District in Manado, North Sulawesi, Indonesia

100.000 population. Virus dengue (DENV) bertahan hidup di alam oleh dua mekanisme, oleh transmisi
horisontal melalui vertebrata yang terinfeksi dan nyamuk, dan dengan vertikal (transovarial) transmisi
dalam nyamuk. Transmisi transovarial diasumsikan sebagai aspek penting dalam memelihara DENV saat
epidemi berlangsung, namun hal ini belum diteliti di Malalayang, Manado. Pengetahuan tentang infeksi
virustransovarial Aedes sp Dengue diperlukan dalam upaya untuk mencegah dan mengendalikan penyakit
DBD
Tujuan: Untuk membuktikan adanya penularan virus Dengue melalui Ae. aegypti dan Ae. albopictus dan
hubungannya dengan kejadian DBD di Malalayang kabupaten di Manado, Sulawesi Utara, Indonesia.
Metode: Metode penelitian ini adalah penelitian observasional analitik dengan desain cross-sectional.
Sampel penelitian adalah nyamuk Ae. aegypti dan Ae. albopictus generasi F1 yang belum pernah menghisap
darah dari hasil pemasangan ovitrap di 5 desa terpilih berdasarkan jumlah kasus dengue di Kabupaten
Malalayang. Data sekunder pasien DBD dari Kabupaten Malalayang diperoleh dari Dinas Kesehatan
Manado dan Puskesmas pada tahun 2010. Keberadaan antigen dengue pada sediaan head squash
sampel nyamuk dideteksi menggunakan antibodi monoklonal antidengue (DSSE10) berdasarkan teknik
imunositokimia streptavidin biotin proxidase complex untuk membuktikan adanya transmisi transovarial
virus dengue dan mendapatkan data indeks transmisi transovarial. Uji Exact Fisher dan korelasi Pearson
digunakan untuk menganalisis data tersebut.
Hasil: Penularan transovarial virus Dengue pada nyamuk Aedes sp ditemukan dari 5 desa di kabupaten
Malalayang dengan index transmisi transovarial (ITT) berkisar 6,1% -17,1%. Uji statistik menunjukkan
positive rates Ae. aegypti terhadap virus dengue secara signifikan lebih tinggi daripada nyamuk Ae.
albopictus (P<0.005) serta tidak ada hubungan yang signifikan secara statistik (p-value 0,528) antara ITT
dengue virus dan angka insidensi (IR) DBD di Kabupaten Malalayang.
Simpulan: Penelitian ini membuktikan adanya transmisi transovarial virus dengue pada nyamuk Aedes sp
di kabupaten Malalayang, dan ITT Ae. aegypti lebih tinggi dibandingkan dengan Ae. albopictus, serta tidak
ada hubungan yang signifikan antara ITT dan angka insidensi DBD.
Kata Kunci: DBD, transmisi transovarial, Ae. aegypti, Ae. albopictus

INTRODUCTION
Dengue Hemorrhagic Fever (DHF) is a
disease transmitted through female Aedes sp.
The disease become one of the most serious
health problem in Indonesia and often cause
an outbreak which leading to death. It was first
discovered in Manado, North Sulawesi in 1973
then spread to various areas. Incidence Rate
(IR) per 100,000 population per year in North
Sulawesi over the last 5 years (2005-2009) was
increased more than 40/100,000 population per
year. Dengue cases in province reported highest
in February 2010 with 615 cases1.
In January 2010, dengue cases were reported
in Manado, followed in Minsel, Minut, Bitung,
Minahasa and Sangihe regencies. Based on the
surveillance report activities in Manado Health

Office, the highest DHF cases was reported in

Malalayang district with 211 cases in 2010 with
incidence rate of 328 per 100,00 population2.
Effort to maintain hygiene in order to
eradicate DHF has actually been declared by
the Mayor of Manado since 1998 through
Clean Friday Movement, followed by Clean
and Green City in 2002 and JUMPA BERLIAN
(clean environment in Friday morning) in
2006. The aim of those actions is to create
a conductive environment and indirectly to
suppress the occurrence DHF cases in Manado.
Mayor of Manado has also issued a form letter
No:440/D.02/75/I/2006 on eradication and DHF
prevention. However, the local government
programs were not optimally realized by both
health personnel and the community3.
Ae.aegypti mosquitoes, are major vector

88

TMJ Vol. 01 No.02 87-95

borne disease for DHF. The mosquitoes are

very anthropophilic and live closed to humans.
Another species of Aedes, Ae. albopictus,
is suggested for being a potential vector of
DHF. Morbidity and mortality of dengue virus
infection is influenced by the host immune
status, the mosquito vector density, dengue virus
transmission, malignancy (virulence) of dengue
virus and local geographical conditions4,5. Dengue
virus transmission generally occurs horizontally
from human carrier dengue virus of its vector,
Aedes sp, after propagation in the mosquito and
transmitted to human recipients6. In addition,
vertical transmission (transovarial) dengue virus
by Aedes sp vector as a parent to the ovum (egg)
in the uterus and then propagates in the eggs,
larvae, pupa, and imago (adult). Transovarial
transmission of dengue virus by its vector in
endemic areas could be a causative key which
responsible for the phenomenon of increasing
cases of DHF. The results of observations made
in India, an Ae. albopictus mosquito showed a
high percentage for the vertical transmission of
dengue virus by Indirect Fluorescence Antibody
Test (IFAT) method7.
Studies on natural transovarial transmission
of dengue virus in Indonesia was first reported
byUmniyati (2004)8 in Kelurahan Klitren,
Gondokusuman Sub-district, Yogyakarta based
on Immunohistochemistry Streptavidin Biotin
Peroxides Complex (ISBPC) method. The method
used head squash preparation of one week old
unbloodfed Ae. aegypti female mosquitoes with
transovarial transmission index (TTI) 27.97%,
which is then standardized by Umniyati (2004)9.
Despite its qualitative nature, it is known to
be sensitive, specific, reliable, and valid for
diagnostic purposes of dengue virus infection
in the Ae. aegypti mosquito6. This is more
convenient and can be performed in places with
less laboratories facilities.

89

The purpose of this study is to prove the

existence of transovarial transmission of dengue
virus in Ae. aegypti and Ae. albopictus mosquito
and its relationship with the incidence of dengue in
Malalayang district in Manado, North Sulawesi.

materials AND METHODS

The method of this research was an
observational analytic study with cross-sectional
design to find out the DHF incidence and to
demonstrate the existence of Dengue virus
transovarial transmission in the Ae. aegypti and
Ae. albopictus mosquitoes in DHF endemic area
in the same period.
Research site was in Malalayang district
Manado municipality, a district with highest
endemic cases for DHF in 2010. Five out of
9 villages were taken representing villages
with high case of dengue in 2010. Secondary
data of dengue patient was obtained from
City Health Office and Community Local
Health Centers, Malalayang District in 2010.
This study used ovitraps for eggs collection,
trays and cages for mosquito colonization6
and immunohistochemical streptavidin biotin
peroxidase complec (ISBPC) assay using
monoclonal antibody against dengue virus
(DSSE10) to detect Dengue viralantigen on head
squash of mosquito specimen8,9.
Ovitrap survey was then performed, and then
followed with OI calculation. Eggs were reared
to get F1 for Dengue virus examination with
ISBPC technique. Positive rate of Dengue virus
differences in Ae. aegypti and Ae. albopictus was
analysed using Fishers Exact Test, whereas to
relationship TII and IR DHF was analysed using
Pearson correlation test.

70-9RO1R
Angle M H Sorisi et. al, Transovarial Transmission Index of Dengue Virus on Aedes aegypti and
Aedes albopictus Mosquitoes in Malalayang District in Manado, North Sulawesi, Indonesia

in Malalayang village I (58 houses), Bahu village

(57 houses), Malalayang village II (57 houses),
Ovitrap distribution
was
in each
house
Malalayang
East I village
(57done
houses),
Batukota
on
Malalayang
village II, Bahu village, Malalayang
village
(57 houses).
EastFour
I village,
Batukota
village and
ovitraps
were mounted
per Malalayang
house, two
district
I.
Sampling
refers
to
the
DHF
Guideline
inside the house (bathroom and living room)
and
Survey
of
Entomology
according
to
WHO
2001
the other two were outside the house (in front
10
criteria
. Ovitraps
were
distibuted
Malalayang
and besides
of the
house).
Thus,intotally
1144
village
(58 houses),
village (57 houses),
ovitrapsI were
mountedBahu
attached.
Malalayang village II (57 houses), Malalayang
East I village (57 houses), Batukota village (57
houses).

represent the number of high, medium, and low

endemic areas. Those 5 endemic DHF village
RESULTs AND DISCUSSIONs
were characterized by some garden and forest,
except for densely Bahu village.
Ovitraps were
distributed
in study
Ovitrap
distribution
was
donearea
in based
each
on
the
number
of
DHF
cases
that
occurred
in
the
house on Malalayang village II, Bahu village,
last 3 years East
per Ivillage
Malalayang
district.
Malalayang
village,in Batukota
village
and
The
5
endemic
DHF
villages
that
were
selected
Malalayang district I. Sampling refers to the DHF
represent the
number
of high, medium,
and low
Guideline
Survey
of Entomology
according
to
endemic
areas.
Those
5
endemic
DHF
village
WHO 2001criteria10. Ovitraps were distibuted
were characterized by some garden and forest,
except for densely Bahu village.

Figure1.1.Ovitrap
Ovitrapmounting
mountinglocation
locationininfive
fivevillages
villagesininMalalayang
Malalayangdistrict
district
Figure
Table 1. Frequency distributions of positive Aedes sp ovitrap in Malalayang district Manado
Villages

Ovitrap distribution
I

Malalayang I
Bahu
Malalayang II
East Malalayang I
Batukota

BR
12
9
15
13
12

Note : BR : Bath Room

LR : Living Room
96

LR
17
13
15
12
13

Amount
29
22
30
25
25

F : Front
S : Side

Total
116
114
114
114
114

F
14
19
20
15
22

S
5
17
21
16
21

O
Amount
19
36
41
31
43

Total
116
114
114
114
114

I : Indoor
O : Outdoor

90

TMJ Vol. 01 No.02 87-95

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ble 2. Frequency
ofdistributions
positive and of
negative
sp ovitraps
andsp
ovitrap
index
in index (OI) in
Tabledistributions
2. Frequency
positiveAedes
and negative
Aedes
ovitraps
and(OI)
ovitrap
DHF endemic villages
in Malalayang
Manadodistrict Manado
DHF endemic
villagesdistrict
in Malalayang
Villages

Ovitrap distribution
Ovitrap distribution
I
O
I
O
(+) egg (-) egg(+)Amount
OI (%)
(+) eggOI(-)
OI (%)
egg (-) egg
Amount
(%)egg(+)Amount
egg (-) egg
Amount OI (%)
Malalayang I Malalayang
29 I 87
29116 87 25 11619
2597
19116 97 16.3 116
16.3
Bahu
92
Bahu 22
22114 92 19.3 11436 19.378
36114 78 31.6 114
31.6
Malalayang II Malalayang
30 II 84
30114 84 26.3 11441 26.373
41114 73 35.9 114
35.9
East MalalayangEast
I Malalayang
25
89I
25114 89 21.9 11431 21.983
31114 83 27.2 114
27.2
Batukota
25
89
Batukota
25114 89 21.9 11443 21.971
43114 71 37.7 114
37.7
Villages

Note : I : Indoor
OI : Ovitrap Index
Note : I : Indoor
OI : Ovitrap Index
O : Outdoor O : Outdoor
Table 3. Positive
Aedes
ovitraps
based
the position
of ovitrapsindoors
and outdoors and outdoors
Table
3. sp
Positive
Aedes
sp on
ovitraps
based on
the position of ovitrapsindoors
in five DHF endemic
in fivevillages
DHF endemic villages
Ovitrap placement
location
Positive
Ovitrap
placement
location
Bathroom
Living room
Front
Besides
Total

Bathroom
Living room
Front
Besides
Total

61
70
90
80
301

Negative
Positive
225
61
216
70
196
90
206
80
843
301

Ovitrap
(+) % Ovitrap
Attached
Negative
(+) %
21.3
225
24.5
216
31.5
196
28
206
26.3
843

286
21.3
286
24.5
286
31.5
286
28
1144
26.3

Attached
286
286
286
286
1144

Ae.
albopictus
Ae. albopictus
in the
gardens,
liveOI
inso
have
gardens,
less
so have
less
villages
in Jakarta,
isthe
higher
outdoors
(36.4%)
Four ovitraps
were
mounted
house,
two live
Table 2 showed,
positive
ovitrap
were per
Table
2 showed,
positive
ovitrap
were
4
4
with human
contact
than
indoors
(33.5%) .because Ae. aegypti prefer
inside
the
house
(bathroom
andindoors.
livingcontact
room)
and
. with human
nd more at found
outdoors
than
indoors.
Ovitrap
more
at outdoors
than
Ovitrap
lay eggs
outdoors
than
mosquitoes
the other
two
were
outside
the in
house
(in
front
After
collection,
After
the ovistraps
collection,
containing
the indoors.
ovistraps
eggsThis
containing
eggs
tribution outcome
in 5 villages
in DHF
distribution
outcome
in 5endemic
villages
DHF endemic
roletointhe
Dengue
because
andthebesides
of
house).
Thus,
totally
1144
brought
to theplays
brought
laboratory,
then
laboratory,
thevirus
eggstransmission,
were
then the eggs
were
as based on
position
ofthethe
ovitraps
areas
based
on the
position
of the
ovitraps
its
lifeforisobtaining
inside
and
around
the F1
house,
while
ovitraps
mounted
attached.
hatched,the
and reared
hatched,
and
reared
F1for
generation
obtaining
generation
throom, living
room,were
front
and
besides
(bathroom,
living
room,
frontthe
and besides
albopictus
in hatch
the
gardens,
so have
less
Table
positive
adults.were
Aedes spAe.
adults.
eggs
hatch
Aedesinto
splive
eggs
larvae
around
into1larvae
around
1
uses) can be houses)
seen
in the
3. in the
can2Table
beshowed
seen
Table 3.ovitrap
4
contact
with
at outdoors
than indoors.
Ovitrap
to 4 days.
The larvae
to 4 days.
stadium
Thehuman
need
larvaeapproximately
stadium
need approximately
The abovefound
table
shows
the highest
Themore
above
table
shows
the
highest
.
distribution
outcome
in 5Aedes
villages
DHF
7-8endemic
days
7-8 After
days
pupae,
to
become
and thethe
pupae,
pupae
and
needthe
pupae need
centage of positive
Aedes
sp ovitrap
was sp
ininovitrap
percentage
of positive
was to
in become
collection,
ovistraps
containing
eggs
areas ofbased
on the position of the
ovitraps
around
2-3 days
around
to turn
2-3
days
adult
to turn
mosquitoes.
intothen
adult
nt of the houses.
front
the houses.
brought
tointo
the
laboratory,
themosquitoes.
eggs were
living
front
besides
the
Usually,
male mosquitoes
Usually, male
appeared
mosquitoes
faster
appeared
than F1 faster
than
The ovitrap (bathroom,
index
result
inroom,
DHF
The(OI)
ovitrap
index
(OI)endemic
resultand
in DHF
endemic
hatched,
and
reared
for
obtaining
generation
houses)
beisseen
in
the
Table
3. female
ones. Once
female
all Aedes
ones.
the mosquitoes
Once
the
turn
mosquitoes
intolarvae around
turn into
as in Malalayang
district
higher
outdoors
areas
in can
Malalayang
district
is higher
outdoors
adults.
sp
eggsallhatch
into
1
above
tablewith
shows
highest
percentage
ones, theto
adult
next
ones,
process
isnext
to stadium
separate
processneed
the
is toapproximately
separate the
n indoors. These
results
similar
thethe
results
thanThe
indoors.
These
results
similar
withadult
the results
4 days.
Thethe
larvae
9
9
ovitrap
was in front
of
the 7-8
adult
mosquito
adult
by its
mosquito
species,
Ae.
by its
aegypti
species,
or
Ae.
Ae.
aegypti
Ae.
Hasyimi who
of positive
Hasyimi
days
to become
pupae,
and
the
pupaeorneed
conductAedes
research
who sp
conduct
in several
research
in several
houses.
albopictus.
albopictus.
found that
five
is found
endemic
that
villages
fiveadult
endemic
villages
2-3 Itdays
to turn
into
mosquitoes.
ages in Jakarta,
villages
OI isin
higher
Jakarta,
outdoors
OI is higher
(36.4%)
outdoors
(36.4%)It isaround
ovitrap
index
(OI) result
DHF
endemic
were
dominated
were
by dominated
Ae.
aegypti
bymosquitoes,
Ae. appeared
aegypti mosquitoes,
Usually,
male
mosquitoes
faster than
n indoors (33.5%)
thanThe
indoors
because
(33.5%)
Ae.
aegypti
because
prefer
Ae.inaegypti
prefer
areas
inindoors.
Malalayang
district
is higher
outdoors
whereas
Ae. albopictus
whereasones.
Ae.
wasOnce
albopictus
found
butwas
few.
found but
female
all the
mosquitoes
turn few.
into
eggs outdoors
lay
than
eggs
outdoors
This
than
mosquitoes
indoors.
This mosquitoes
than indoors.
These results
similar withMoreover,
the
resultsmostAe.
Moreover,
adultalbopictus
ones,mostAe.
the mosquitoes
nextalbopictus
processdied
ismosquitoes
to separatedied
the
ys role in Dengue
plays
virus
role intransmission,
Dengue
virusbecause
transmission,
because
9
ofand
Hasyimi
after
the
age ofafter
twothe
days,
agewhereas
ofby
two
days,
aegypti
whereas
Ae. aegypti
adult
mosquito
its Ae.
species,
Ae. aegypti
or Ae.
life is insideits
life around
is inside
the
and
house,
around
while
the house,
while
who
conduct
research
in several

91

97

97

70-9RO1R
70-9RO1R

Angle M H Sorisi et. al, Transovarial Transmission Index of Dengue Virus on Aedes aegypti and
Aedes albopictus Mosquitoes in Malalayang District in Manado, North Sulawesi, Indonesia

mosquitoes
ablethat
to five
survive
more villages
than a
albopictus. It were
is found
endemic
mosquitoes were able to survive more than a
week.
Based on this
condition,
were dominated
by Ae.
aegyptiseven-day-old
mosquitoes,
week. Based on this condition, seven-day-old
female
Ae.
aegypti
,
and
two-day-old
whereas Ae. albopictus was found butfemale
few.
female Ae. aegypti , and two-day-old female
Ae.
albopictuswere
in this
study. Thirty
Moreover,
most Ae.selected
albopictus
mosquitoes
died
Ae. albopictuswere selected in this study. Thirty
female
aegypti
were
after theAe.
age
of twomosquitoes
days, whereas
Ae.detected
aegypti
female Ae. aegypti mosquitoes were detected
in
each village
andable
all to
Ae.survive
albopictus
eacha
mosquitoes
were
moreinthan
in each village and all Ae. albopictus in each
village.
Positiveonand
control
mosquitos
week. Based
thisnegative
condition,
seven-day-old
village. Positive and negative control mosquitos
were
taken
from
Laboratory
of
Parasitology,
female Ae. aegypti, and two-day-old female
were taken from Laboratory of Parasitology,
Faculty
of Medicine,
Gadjah
Ae. albopictus
wereUniversitas
selected in
this Mada.
study.
Faculty of Medicine, Universitas Gadjah Mada.
The
presence
of
dengue
antigen
on
head
Thirty female Ae. aegypti mosquitoes were
The presence of dengue antigen on head
squashes
male Ae.
detected of
in intra
each thoracally
village and- infected
all Ae. albopictus
squashes of intra thoracally - infected male Ae.
aegypti
wereand
detected
based
on
in each mosquitoes
village. Positive
negative
control
aegypti mosquitoes were detected based on
ISBPC
technique
monoclonal
mosquitos
were using
takencomercially
from Laboratory
of
ISBPC technique using comercially monoclonal
antibody
against
dengue
as positiveUniversitas
controls.
Parasitology,
Faculty
of Medicine,
antibody against dengue as positive controls.
Negative
control
tissue specimens
Gadjah Mada.
The presence
of dengue without
antigen
Negative control tissue specimens without
primary
usedthoracally
as negative- controls.
on headantibody
squasheswere
of intra
infected
primary antibody were used as negative controls.
Dengue
antigenmosquitoes
was detected
as brownish
male
Ae. aegypti
were
detected
Dengue antigen was detected as brownish
color
in
the
cytoplasm
of
infected
cells
or as
based on ISBPC technique using comercially
color in the cytoplasm of infected cells or as
discrete
brownish
granular
monoclonal
antibody
againstdeposits
dengue asscattered
positive
discrete brownish granular deposits scattered
throughout
brain
tissue
of
positive
controls,
controls. Negative control tissue specimens
throughout brain tissue of positive controls,
and
positive
samples.
Thewere
negative
result
was
without
primary
antibody
used as
negative
and positive samples. The negative result was
shown
as blue or purple colour throughout brain
controls.
shown as blue or purple colour throughout brain
tissue
of negative
controls
and negative
samples
Dengue
antigen
was detected
as brownish
tissue of negative controls and negative samples
(Figure
color in2).the cytoplasm of infected cells or as
(Figure 2).
Dengue
virus detection
of unbloodfed
discrete
brownish
granularresults
deposits
scattered
Dengue virus detection results of unbloodfed
Ae.
aegypti brain
samples
on the
F1 generation
in
throughout
tissue
of positive
controls,
Ae. aegypti samples on the F1 generation in
5
DHF
endemic
villages
can
be seenresult
in Table
and
positive
samples.
The
negative
was
5 DHF endemic villages can be seen in Table
4.
The
result
showed
that
the
highest
TII
in
shown as blue or purple colour throughout brain
4. The result showed that the highest TII in

DHF endemic
areas
was and
in Malalayang
I and
tissue
of negative
controls
negative samples
DHF endemic areas was in Malalayang I and
Malalayang
(Figure
2). II with TII 20%, whereas, the the
Malalayang II with TII 20%, whereas, the the
lowest
TII was
in detection
East Malalayang
I. unbloodfed
Dengue
virus
results of
lowest TII was in East Malalayang I.
Ae. aegypti samples on the F1 generation in 5

Positive Dengue virus

Positive Dengue virus

Negative Dengue virus

Negative Dengue virus

Positive control
Negative control
Positive control
Negative control
Figure 2. Head squashes of immunohistochemical
Figure 2.
2. Head
squashes
immunohisto
Head squashes
preparation
ofof immunohistochemical
a ofpositive
sample
chemical
preparation
of
a showing
positive
preparation
of
a
positive
sample
and a positive control (left)
sample
and
a
positive
control
(left)
and
a
positive
control
(left)
showing
positive reaction as brownish color
showing
positive
reaction
as
brownish
positive
reaction
as
brownish
color
in the cytoplasm of infected cells and
color
thebrownish
cytoplasm
of infected
cells
in
theincytoplasm
of infected
cells
and
descrete
colour
deposits
and
descrete
brownish
colour
deposits
descrete
brownish
colour
throughout brain tissues, and negative
throughout
brain
tissues,
and
results
(right)
were
shown
asnegative
blue or
results
(right)
were
shown
as blue
or
purple colour throughout brain
tissues
colour
throughout
brain
tissues
purple
of a negative sample and a negative
of a negative sample and a negative
control.
control.

Table 4. The result of microscopic examination of dengue antigen on head squashes

Table 4. of
Theunbloodfed
result of microscopic
of dengue
antigenaton400x
headand
squashes
Ae. aegyptiexamination
samples on the
F1 generation
1000x
of unbloodfed Ae.
aegypti
samples on the F1 generation
400xmonoclonal
and 1000x
magnification
based
on immunohistochemical
assays at
using
magnification
based
on (DSSE10)
immunohistochemical
assays in
using
monoclonal
antibody
against
dengue
as primary antibody
5 DHF
endemic
antibodyin against
dengue
(DSSE10)
as primary antibody in 5 DHF endemic
villages
Malalayang
District
year 2011
villages in Malalayang District year 2011
No
Villages
Sample
No
Villages
Sample
1 Malalayang I
30
1 Malalayang I
30
2 Bahu
30
2 Bahu
30
3 Malalayang II
30
3 Malalayang II
30
4 East Malalayang I
30
4 East Malalayang I
30
5 Batukota
30
5 Batukota
30
TTI= Transovarial transmission index
TTI= Transovarial transmission index
98
98

Total
Total
Positive
Positive
6
6
3
3
6
6
2
2
3
3

TTI (%)
TTI (%)
20
20
10
10
20
20
6.7
6.7
10
10

92

TMJ Vol. 01 No.02 87-95

$QJOH0+6RULVLHWDO7UDQVRYDULDO7UDQVPLVVLRQ,QGH[RI'HQJXH9LUXVRQ$HGHVDHJ\SWLDQG
$HGHVDOERSLFWXV0RVTXLWRLQ0DODOD\DQJ'LVWULFWLQ0DQDGR1RUWK6XODZHVL,QGRQHVLD

Table 5. The result of microscopic examination of dengue antigen on head squashes of

unbloodfed Ae. albopictus samples on the F1 generation at 400x and 1000x
magnification based on the immunohistochemical assays using monoclonal
antibody against dengue (DSSE10) as primary antibody in 5 DHF endemic
villages in Malalayang District year 2011
No

Villages

1
2
3
4
5

Malalayang I
Bahu
Malalayang II
East Malalayang I
Batukota

Total
Positive
0
0
0
0
1

Sample
15
0
5
3
22

TTI (%)
0
0
0
0
4.5

Table 6. Dengue virus TII difference in Ae. aegypti and Ae. albopictus mosquitoes
No

Villages

1
2
3
4
5

Malalayang I
Bahu
Malalayang II
East Malalayang I
Batukota

TTI (%)
Ae. Aegypti
20
10
20
6.7
10

TTI (%)
Ae. Albopictus
0
0
0
0
4.5

Table 7. Dengue virus positive rate in Ae. aegypti and Ae. albopictus mosquitoes
Mosquito Spesies
Ae. Aegypti
Ae. Albopictus

Negatif Dengue
virus
130
44

Positif Dengue
virus
20
1

p value
0,00
(p<0,05)

Table 8. The result of microscopic examination of dengue antigen on head squashes

of unbloodfed Aedes sp samples on the F1 generation at 400x and 1000x
magnification based on the immunohistochemical assays using monoclonal
antibody against dengue (DSSE10) as primary antibody in 5 DHF endemic
villages in Malalayang District year 2011
No
1
2
3
4
5

93

Villages
Malalayang I
Bahu
Malalayang II
East Malalayang I
Batukota

Ae. Aegypti
30
30
30
30
30

Ae. Albopictus
15
0
5
3
22

Total Aedes spp Positive

45
6
30
3
35
6
33
2
52
4

TTI (%)
13.3
10
17.1
6.1
7.7

99

70-9RO1R

Angle M H Sorisi et. al, Transovarial Transmission Index of Dengue Virus on Aedes aegypti and
Aedes albopictus Mosquitoes in Malalayang District in Manado, North Sulawesi, Indonesia

Table 9. Dengue virus positive rate in Ae. aegypti and Ae. albopictus mosquitoes
Villages
Malalayang I
Bahu
Malalayang II
East Malalayang I
Batukota

IR year 2010
72.5
46.2
31.2
25.3
25.8

TII (%) Aedes sp.

13.3
10
17.1
6.1
7.7

p value
0.528
(p>0.05)

DHF endemic villages can be seen in Table 4. The

Dengue
virus
detection
results
unbloodfed
result
showed
that
the highest
TII inofDHF
endemic
Ae.
albopictus
samples
on
the
F1
generation
in
areas was in Malalayang I and Malalayang II with
5 DHF
villages
can
be seen
in Table
5,
TII
20%,endemic
whereas,
the the
lowest
TII was
in East
whereas theI. differences between Ae. aegypti
Malalayang
andDengue
Ae. albopictus
Dengue results
virus TTI
be seen
virus detection
of can
unbloodfed
in
Table 6, and
the differences
between Ae.
Ae.the
albopictus
samples
on the F1 generation
in
aegypti
and Ae. albopictus
Dengue
virus
positive
5 DHF endemic
villages can
be seen
in Table
5,
rate
can
be
seen
in
the
Table
7.
whereas the differences between Ae. aegypti
5 showed that
transovarial
andTable
Ae. albopictus
Dengue
virus TTItransmission
can be seen
of
dengue
virus
in
Ae.
albopictusonly
occuredAe.
in
in the Table 6, and the differences between
Batukota
with
of 4.5%.
The result
showed
aegypti and
Ae.TTI
albopictus
Dengue
virus positive
that
TTI be
of seen
Ae. aegypti
was significantly
higher
rate can
in the Table
7.
thanTable
Ae. albopictus.
Thetransovarial
results were
similiar to
5 showed that
transmission
those
performed
byAe.
Wanti
(2010) inonly
Kupang,
that
of dengue
virus in
albopictus
occured
found
that with
Ae. aegypti
TTI (20.14%)
wasshowed
higher
in Batukota
TTI of 4.5%.
The result
11
than
Ae. albopictus
(8.33%)
that TTI
of Ae. aegypti
was .significantly higher
of Fisher`s
Exact were
Test confirmed
thanThe
Ae.result
albopictus.
The results
similiar to
that
the
positive
rates
of
Ae.
aegypti
was
those performed by Wanti (2010) in Kupang, that
significantly
higher
than TTI
Ae. (20.14%)
albopictus
(Table
7).
found
that Ae.
aegypti
was
higher
11
BasedAe.
onalbopictus
the results(8.33%)
of microscopic
examination
than
.
of the
the Test
highest
TTI in
The Aedessp
result ofmosquito,
Fisher`s Exact
confirmed
DHF
areasrates
was inof Malalayang
that endemic
the positive
Ae. aegyptiII with
was
TII
17.1%,
followed
by
Malalayang
I
13.3%,
Bahu
significantly higher than Ae. albopictus (Table 7).
10%,
7.7%
and East Malalayang
Based Matukota
on the results
of microscopic
examinationI
village
8).
of the 6.1%
Aedes(Table
sp mosquito,
the highest TTI in
assess the
between IIAedes
DHFToendemic
areasrelationship
was in Malalayang
with
spp
with
DHF
incidence
rate
(IR),
Pearson
TII 17.1%, followed by Malalayang I 13.3%, Bahu
correlation
statistic
test East
wasMalalayang
done and p-value
10%, Matukota
7.7% and
I village
=6.1%
0.528
was
obtained.
It
indicates
that
there
was
(Table 8).
no significant
correlation
between
the Dengue
To assess the
relationship
between
Aedes
virusTTIon
Aedes
spp
mosquitoes
with
the
IR of
spp with DHF incidence rate (IR), Pearson

no significant correlation between the Dengue

DHF
villages
virus in
TTI5on
Aedes in
sp Malalayang
mosquitoes district,
with thewhile
IR of
the
value
of
closeness
of
the
relationship
with
DHF in 5 villages in Malalayang district, whiler
=the
0.38
indicates
the correlation
was not strong.
value
of closeness
of the relationship
with r
This
is
possible
because
of
the
ease
transport
= 0.38 indicates the correlation was not strong.
between
regionbecause
led to ofincrease
This is possible
the easepopulation
transport
mobility,
of dengue
viruses
between allowing
region the
led spread
to increase
population
from
other
regions.
this study,
patients
with
mobility,
allowing
theInspread
of dengue
viruses
DHF
case
data
were
not
classified
by
patient
from other regions. In this study, patients with
age,
there
waswere
the posibility
of cases
DHF so
case
data
not classified
by derived
patient
from
other
areas
because
of
the
high
population
age, so there was the posibility of cases derived
mobility.
In areas
contrast
study of
result
by Sucipto
from other
because
the done
high population
(2009),
casestudy
limited
ondone
children
aged
mobility.patient
In contrast
result
by Sucipto
under
assuming
the caseoncomes
fromaged
the
(2009),five,
patient
case limited
children
12
local
undersite
five,. assuming the case comes from the

correlation statistic test was done and p-value =

0.528 was obtained. It indicates that there was

endemic and sporadic DHF villages.

100

local site12.

CONCLUSION AND
SUGGESTION

CONCLUSION

This study found the existence of Dengue

virus
transovarial
withthe
different
TTI inofeach
DHF
This
study found
existence
Dengue
endemic
villages with
ranged
from TTI
6.1%in to
17.1%.
virus transovarial
different
each
DHF
The
Dengue
virus
transovarial
transmission
endemic villages ranged from 6.1% to 17.1%.
was
between
Ae. aegypti
and Ae.
The different
Dengue virus
transovarial
transmission
albopictus,
withbetween
TTI in Ae.Ae.
albopictus
was
different
aegyptiwas
andlower
Ae.
than
Ae. aegyptis.
test result
showed
albopictus,
with TTI Statistic
in Ae. albopictus
was
lower
no
correlation
between
the Aedessp
thansignificant
Ae. aegyptis.
Statistic
test result
showed
mosquito
Dengue
virus
TTI
with
DHF
IR
in
5 DHF
no significant correlation between the Aedessp
endemic
Malalayang
mosquitovillages
Dengueinvirus
TTI with district.
DHF IR inFurther
5 DHF
research
needs
to
be
done
with
more
number
of
endemic villages in Malalayang district. Further
villages,
ordertotobe
get
complete
data number
from each
researchinneeds
done
with more
of
endemic
and
sporadic
DHF
villages.
villages, in order to get complete data from each

94

TMJ Vol. 01 No.02 87-95

ACKNOWLEDGEMENTS
The author would like to thank the Chairman
of Basic Medical Science and Biomedical Program
Study, Head of Parasitology Department,
including Joko Trimuratno and Suprihatin for
their helpful assistance. Our gratitude is also
for Manado Head of Health Office and Head of
Bahu and Minanga CHCs for the permission and
supports during the study.

REFERENCES
1. Rondonuwu MR. Kebijakan Pengendalian DBD
di Provinsi Sulawesi Utara. Dalam Seminar
Penanggulangan Demam Berdarah Dengue
10 April. Fakultas Kesehatan Masyarakat
Universitas Samratulangi. Manado, 2010.
2. Dinkes Kota Manado, Data Surveilans DBD.
Manado, 2010.
3. Suwarja. Kondisi Sanitasi Lingkungan Dan
Vektor Dengue Demam Berdarah Pada Kasus
Penyakit DBD di Kecamatan Tikala Kota
Manado. [Tesis]. Yogyakarta, Ilmu Kesehatan
Masyarakat: UGM, 2007.
4. WHO. Demam Berdarah Dengue. Diagnosis,
Pengobatan, Pencegahan, dan Pengendalian.
Alih bahasa Monica Ester. Jakarta: EGC,
1999.
5. Depkes RI. Pencegahan dan Pemberantasan
Demam Berdarah Dengue. Ditjen PPM & PLP
Depkes RI. Jakarta, 2005.
6. Mardihusodo SJ, Satoto TBT, Mulyaningsih
B, Umniyati SR & Ernaningsih. Bukti Adanya
Penularan Virus Dengue Secara Transovarial
Pada Nyamuk Aedes aegypti Di Kota

95

Yogyakarta. Simposium Nasional Aspek

Biologi Molekuler, Patogenesis, Manajemen
dan Pencegahan KLB, 16 Mei di Pusat Studi
Bioteknologi UGM. Yogyakarta, 2007.
7. Angel & Joshi. Distribution And Seasonality
Of Vertically TransmTIIed Dengue Viruses
In Aedes Mosquitoes In Arid And Semi-Arid
Areas Of Rajasthan, India. J Vector Borne
Dis45. Desert Medicine Research Centre,
Indian Council of Medical Research, Jodhpur,
India, 2008;56-9.
8. Umniyati SR. Preliminary investigation on the
transovarial transmission of Dengue Virus in
the population of Aedes aegypti in the well.
Dalam Seminar Hari Nyamuk IV, 21 Agustus.
Surabaya, 2004.
9. Umniyati SR.
Standardization of
immunocytochemical method for the
diagnosis of dengue viral infection in Aedes
aegypti Linn mosquitoes (Diptera: Culicidae).
BIK, 2009;41:1-10
10. WHO. Prevention and Control of Dengue and
Dengue Haemorrhagic Fever, Comprehensif
Guidelines. Diterjemahkan oleh Palupi W.
Jakarta: EGC, 2001.
11. Wanti. Demam Berdarah Degue di Kota
Kupang: Kodisi Iklim, Status Etomologi
dan bukti adanya Infeksi transovarial virus
Dengue pada nyamuk Aedes aegypti dan
Aedes albopictus. [Tesis]. Yogyakarta, Ilmu
Kedokteran Tropis: UGM, 2010.
12. Sucipto CD. Deteksi Trasmisi Transovarial
Virus Dengue pada nyamuk Aedes aegypti
jantan dan betina serta hubungannya dengan
Incidence Rate Demam Berdarah Dengue
di Kota Pontianak. [Tesis]. Yogyakarta, Ilmu
Kesehatan Tropis: UGM, 2009.