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
1

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. Fisher’s 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. aegypti’s 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 Office, the highest DHF cases was reported in

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
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 Fisher’s Exact Test, whereas to
relationship TII and IR DHF was analysed using
Pearson correlation test.
70-‡9RO‡1R‡
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

represent the number of high, medium, and low in Malalayang village I (58 houses), Bahu village
endemic areas. Those 5 endemic DHF village (57 houses), Malalayang village II (57 houses),
RESULTs AND DISCUSSIONs
were characterized by some garden and forest, Ovitrap distribution
Malalayang East I village was
(57done in each
houses), house
Batukota
except for densely Bahu village. on Malalayang
village village II, Bahu village, Malalayang
(57 houses).
Ovitraps were
Ovitrap distributed
distribution wasin study
donearea in based
each EastFour
I village, Batukota
ovitraps village and
were mounted per Malalayang
house, two
on the number of DHF cases that occurred
house on Malalayang village II, Bahu village, in the district I. Sampling refers to the
inside the house (bathroom and living room)DHF Guideline
and
last 3 years East
Malalayang per Ivillage
village,in Batukota
Malalayang district.
village and Survey of Entomology according to
the other two were outside the house (in front WHO 2001
The 5 endemic DHF villages that were selected
Malalayang district I. Sampling refers to the DHF criteria 10
and besides . Ovitraps
of thewere distibuted
house). Thus,intotally
Malalayang
1144
represent the
Guideline number
Survey of high, medium,
of Entomology and low
according to village
ovitrapsI were
(58 houses),
mountedBahu village (57 houses),
attached.
endemic areas. Those 5 endemic DHF
WHO 2001criteria10. Ovitraps were distibuted village Malalayang village II (57 houses), Malalayang
were characterized by some garden and forest, East I village (57 houses), Batukota village (57
except for densely Bahu village. houses).

Figure1.1.Ovitrap
Figure Ovitrapmounting
mountinglocation
locationininfive
fivevillages
villagesininMalalayang
Malalayangdistrict
district

Table 1. Frequency distributions of positive Aedes sp ovitrap in Malalayang district Manado

Villages Ovitrap distribution
I O
BR LR Amount Total F S Amount Total
Malalayang I 12 17 29 116 14 5 19 116
Bahu 9 13 22 114 19 17 36 114
Malalayang II 15 15 30 114 20 21 41 114
East Malalayang I 13 12 25 114 15 16 31 114
Batukota 12 13 25 114 22 21 43 114
Note : BR : Bath Room F : Front I : Indoor
LR : Living Room S : Side O : Outdoor

96
90
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ble 2. Frequency
Tabledistributions
2. Frequencyofdistributions
positive and of
negative
positiveAedes sp ovitraps
and negative andsp
Aedes ovitrap index
ovitraps and(OI) in index (OI) in
ovitrap
DHF endemic villages in Malalayang
DHF endemic villagesdistrict Manadodistrict Manado
in Malalayang

Villages Villages Ovitrap distribution

Ovitrap distribution
I I O O
(+) egg (-) egg(+)Amount OI (%)
egg (-) egg (+) eggOI(-)
Amount (%)egg(+)Amount OI (%)
egg (-) egg Amount OI (%)
Malalayang I Malalayang
29 I 87 29116 87 25 11619 2597 19116 97 16.3 116 16.3
Bahu Bahu 22 92 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
Batukota 89 25114 89 21.9 11443 21.971 43114 71 37.7 114 37.7
Note : I : Indoor OI : Ovitrap Index
Note : I : Indoor OI : Ovitrap Index
O : Outdoor O : Outdoor

Table 3. Positive Aedes

Table 3. sp ovitraps
Positive based
Aedes sp on the position
ovitraps of ovitrapsindoors
based on and outdoors and outdoors
the position of ovitrapsindoors
in five DHF endemic
in fivevillages
DHF endemic villages

Ovitrap placement location

Ovitrap Positive
placement location Negative
Positive Ovitrap (+) % Ovitrap
Negative Attached
(+) % Attached
Bathroom Bathroom 61 225
61 21.3
225 286
21.3 286
Living room Living room 70 216
70 24.5
216 286
24.5 286
Front Front 90 196
90 31.5
196 286
31.5 286
Besides Besides 80 206
80 28
206 286
28 286
Total Total 301 843
301 26.3
843 1144
26.3 1144

Table 2 showed, Four ovitraps

Table positive
2 showed, were
ovitrapmountedwere per
positive Ae. albopictus
house,
ovitrap two live
were Ae. albopictus
in the
villages gardens,
in Jakarta, liveOI insoisthe
havegardens,
higher less so have
outdoors less
(36.4%)
inside
nd more at foundoutdoors the
morehouse
than (bathroom
indoors.
at outdoors Ovitrap
than livingcontact
andindoors. room) with human
and
Ovitrap contact
than . with human
indoors
4
(33.5%) .because Ae. aegypti prefer
4

the other
tribution outcome two
in 5 villages
distribution were
outcome outside
in DHFin 5endemicthe in
villages house (in
DHF endemicfront
After lay eggs
collection, After outdoors
the ovistraps
collection, than
the indoors.
containing eggsThis
ovistraps mosquitoes
containing eggs
as based on andthebesides
areas position
based of
on the house).
ofthethe Thus,
ovitraps
position of thetotally
brought
ovitrapsto theplays
1144 brought roletointhe
laboratory, Dengue
then thevirus
laboratory, eggstransmission,
were
then the eggs because
were
throom, living ovitraps
room,were
(bathroom, front mounted
livingand attached.
besides
room, frontthe hatched,the
and besides its
and reared lifeforisobtaining
hatched, inside
and rearedand F1for around
generation
obtainingthe F1house, while
generation
uses) can be houses)
seenTable can2Table
in the beshowed3. in the
seen positive
Table 3.ovitrap Aedes spAe.
adults.were albopictus
adults.
eggs hatch
Aedesinto splive
eggs in hatch
larvae the gardens,
aroundinto1larvaeso have
around less
1
The abovefound Themore
table shows
above at outdoors than indoors.
the highest
table shows the Ovitrap
to 4 days.
highest contact
The larvae
to 4 days. with
stadium Thehuman
need
larvaeapproximately
stadium
4
. need approximately
distribution
centage of positive
percentage Aedes outcome
sp ovitrap
of positive in 5Aedes
villages
was sp DHF
7-8endemic
ininovitrap days
was to in become
7-8 After
days
pupae,to become
and thethe
collection, pupae,
pupae and
needthe
ovistraps pupae need
containing eggs
areas ofbased
nt of the houses.
front on the position of the
the houses. ovitraps
around 2-3 daysaround
to turn
brought 2-3
tointo
days
the adultto turn
mosquitoes.
laboratory, intothen
adultthemosquitoes.
eggs were
The ovitrap (bathroom,
index
The(OI)ovitrap living
result inroom,
index DHF front
resultand
(OI)endemic in DHFbesides
Usually,
endemicthe
male mosquitoes
Usually, male
hatched, andappeared
mosquitoes
reared forfaster appeared
than F1 faster
obtaining than
generation
houses)
as in Malalayang
areas in can
district beisseen
Malalayang higher in the Table
outdoors
district 3. female
is higher ones. Once
outdoors female
adults.all Aedes
ones.
the mosquitoes
Once
sp eggsallhatchthe
turnmosquitoes
intolarvae around
into turn into 1
thanThe
n indoors. These above
results
indoors. tablewith
similar
These showsthethe
results highest
results
similar percentage
withadult ones, theto
the results adult
next ones,
4 days.process
Thethe isnext
larvae to stadium
separate
processneed the
is toapproximately
separate the
Hasyimi who
9
of positive
conductAedes
Hasyimi 9
who sp
research ovitrap
conduct was in front
in several
research adult of the 7-8
mosquito
in several adult
by its
daysmosquito
species,
to become Ae.
by its
aegypti
species,
pupae, or
andAe.
Ae.
theaegypti
pupaeorneedAe.
houses.
ages in Jakarta,
villages
OI isin higher
Jakarta, outdoors
OI is higher
(36.4%)outdoors albopictus.
(36.4%)It isaround
albopictus.
found that 2-3 Itdays
five
is found
endemic
to turn that villages
fiveadult
into endemic villages
mosquitoes.
thanThe
n indoors (33.5%) ovitrap
indoors
because index
(33.5%)
Ae. (OI) result
aegypti
because Ae.inaegypti
prefer DHF
wereendemic
dominated
prefer were by dominated
Usually, Ae.
male aegypti bymosquitoes,
mosquitoes Ae. appeared
aegypti mosquitoes,
faster than
eggs outdoors areas
lay than
eggs inindoors.
Malalayang
outdoors This
than district
mosquitoes
indoors.is higher outdoors
whereas
This mosquitoes Ae. albopictus
whereasones.
female Ae.
wasOnce
albopictus
found all thebutwas few.found but
mosquitoes turn few.
into
than indoors.
ys role in Dengue
plays virus These results
role intransmission,
Dengue similar withMoreover,
virusbecause
transmission, the resultsmostAe.
because Moreover,
adultalbopictus
ones,mostAe.
the mosquitoes
nextalbopictus
processdied ismosquitoes
to separatedied the
ofand
life is insideits Hasyimi
life around
is inside
9
the
who and house,
around
conduct while after
the house,
research the
while
in several age ofafter
twothe
adult days,
agewhereas
mosquito ofby twoits Ae.
days, aegypti
species,whereas Ae. aegypti
Ae. aegypti or Ae.

91 97 97
 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
70-‡9RO‡1R‡
70-‡9RO‡1R‡

mosquitoes
albopictus. It wereis foundablethatto five
survive more villages
endemic than a DHF endemic
tissue areas
of negative was and
controls in Malalayang I and
negative samples
mosquitoes were able to survive more than a DHF endemic areas was in Malalayang I and
week. Based on this
were dominated by Ae. condition,
aegyptiseven-day-old
mosquitoes, Malalayang
(Figure 2). II with TII 20%, whereas, the the
week. Based on this condition, seven-day-old Malalayang II with TII 20%, whereas, the the
female
whereas Ae. albopictus was found butfemale
Ae. aegypti , and two-day-old few. lowest TII was
Dengue in detection
virus East Malalayang I. unbloodfed
results of
female Ae. aegypti , and two-day-old female lowest TII was in East Malalayang I.
Ae. albopictuswere
Moreover, most Ae.selected
albopictus in this study. Thirty
mosquitoes died Ae. aegypti samples on the F1 generation in 5
Ae. albopictuswere selected in this study. Thirty
female
after theAe. ageaegypti
of twomosquitoes
days, whereas were Ae.detected
aegypti
female Ae. aegypti mosquitoes were detected
in each village
mosquitoes were andableall to
Ae.survive
albopictus
moreinthan eacha
in each village and all Ae. albopictus in each
village. Positiveonand
week. Based thisnegative control
condition, mosquitos
seven-day-old
village. Positive and negative control mosquitos
were taken from Laboratory
female Ae. aegypti, and two-day-old female of Parasitology,
were taken from Laboratory of Parasitology,
Faculty of Medicine,
Ae. albopictus wereUniversitas
selected in Gadjah
this Mada.
study.
Faculty of Medicine, Universitas Gadjah Mada.
The presence of dengue
Thirty female Ae. aegypti mosquitoes were antigen on head
The presence of dengue antigen on head
squashes
detected of in intra
each thoracally
village and- infected male Ae.
all Ae. albopictus Positive Dengue virus Negative Dengue virus
squashes of intra thoracally - infected male Ae. Positive Dengue virus Negative Dengue virus
aegypti
in each mosquitoes
village. Positive wereand detected
negative based
controlon
aegypti mosquitoes were detected based on
ISBPC technique
mosquitos were usingtakencomercially monoclonal
from Laboratory of
ISBPC technique using comercially monoclonal
antibody
Parasitology,against dengue
Faculty as positiveUniversitas
of Medicine, controls.
antibody against dengue as positive controls.
Negative
Gadjah Mada. control tissue specimens
The presence of dengue without
antigen
Negative control tissue specimens without
primary
on headantibody
squasheswere usedthoracally
of intra as negative- controls.
infected
primary antibody were used as negative controls.
male Dengue antigenmosquitoes
Ae. aegypti was detected as brownish
were detected
Dengue antigen was detected as brownish
color in the cytoplasm of infected
based on ISBPC technique using comercially cells or as
color in the cytoplasm of infected cells or as
discrete
monoclonal brownish
antibody granular
againstdeposits
dengue asscattered
positive Positive control Negative control
discrete brownish granular deposits scattered Positive control Negative control
throughout brain tissue of
controls. Negative control tissue specimenspositive controls,
throughout brain tissue of positive controls, Figure 2. Head squashes of immunohistochemical
and positive
without primary samples.
antibody Thewere
negative
used as result
negativewas Figure 2.
2. Head squashes
Head squashes
preparation a ofpositive
immunohisto
ofof immunohistochemical
sample
and positive samples. The negative result was chemical
preparation preparation
of a of a showing
positive positive
sample
shown
controls.as blue or purple colour throughout brain and a positive control (left)
shown as blue or purple colour throughout brain sample
and a and
positive a positive
control control
(left)
positive reaction as brownish color (left)
showing
tissue of negative
Dengue antigen controls and negative
was detected samples
as brownish
tissue of negative controls and negative samples showing
positive positive
reaction reaction
as as
brownishbrownish
in the cytoplasm of infected cells and color
(Figure
color in2).the cytoplasm of infected cells or as color
in theincytoplasm
thebrownish
cytoplasm of infected
of infected cells cells
and
(Figure 2). descrete colour deposits
Dengue
discrete virus detection
brownish granularresults of unbloodfed
deposits scattered and descrete
descrete brownish
brownish colour
colour
throughout brain tissues, and negativedeposits
Dengue virus detection results of unbloodfed throughout
results brain
(right) weretissues,
shown and
asnegative
blue or
Ae. aegypti brain
throughout samples tissueon the F1 generation
of positive controls, in
Ae. aegypti samples on the F1 generation in results (right) were shown
purple colour throughout brain as blue or
tissues
5
andDHF endemic
positive villages
samples. Thecan be seenresult
negative in Table
was purple colour throughout brain tissues
5 DHF endemic villages can be seen in Table of a negative sample and a negative
4. The result showed that the
shown as blue or purple colour throughout brain highest TII in of a negative sample and a negative
control.
4. The result showed that the highest TII in control.
Table 4. The result of microscopic examination of dengue antigen on head squashes
Table 4. of
Theunbloodfed
result of microscopic
Ae. aegyptiexamination of dengue
samples on the antigenaton400x
F1 generation headand
squashes
1000x
of unbloodfed Ae.
magnification aegypti
based samples on the F1 generation
on immunohistochemical assays at 400xmonoclonal
using and 1000x
magnification
antibody based
against on (DSSE10)
dengue immunohistochemical assays in
as primary antibody using monoclonal
5 DHF endemic
antibodyin against
villages dengue
Malalayang (DSSE10)
District as primary antibody in 5 DHF endemic
year 2011
villages in Malalayang District year 2011
Total
No Villages Total
No Villages Sample Positive TTI (%)
Sample Positive TTI (%)
1 Malalayang I 30 6 20
1 Malalayang I 30 6 20
2 Bahu 30 3 10
2 Bahu 30 3 10
3 Malalayang II 30 6 20
3 Malalayang II 30 6 20
4 East Malalayang I 30 2 6.7
4 East Malalayang I 30 2 6.7
5 Batukota 30 3 10
5 Batukota 30 3 10
TTI= Transovarial transmission index
TTI= Transovarial transmission index

92
98
98
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

Total
No Villages
Sample Positive TTI (%)
1 Malalayang I 15 0 0
2 Bahu 0 0 0
3 Malalayang II 5 0 0
4 East Malalayang I 3 0 0
5 Batukota 22 1 4.5

Table 6. Dengue virus TII difference in Ae. aegypti and Ae. albopictus mosquitoes

TTI (%) TTI (%)

No Villages
Ae. Aegypti Ae. Albopictus
1 Malalayang I 20 0
2 Bahu 10 0
3 Malalayang II 20 0
4 East Malalayang I 6.7 0
5 Batukota 10 4.5

Table 7. Dengue virus positive rate in Ae. aegypti and Ae. albopictus mosquitoes

Negatif Dengue Positif Dengue p value

Mosquito Spesies
virus virus
Ae. Aegypti 130 20 0,00
Ae. Albopictus 44 1 (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 Villages Ae. Aegypti Ae. Albopictus Total Aedes spp Positive TTI (%)
1 Malalayang I 30 15 45 6 13.3
2 Bahu 30 0 30 3 10
3 Malalayang II 30 5 35 6 17.1
4 East Malalayang I 30 3 33 2 6.1
5 Batukota 30 22 52 4 7.7

93 99
70-‡9RO‡1R‡ 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 IR year 2010 TII (%) Aedes sp. p value

Malalayang I 72.5 13.3 0.528
Bahu 46.2 10 (p>0.05)
Malalayang II 31.2 17.1
East Malalayang I 25.3 6.1
Batukota 25.8 7.7

DHF endemic villages can be seen in Table 4. The no significant correlation between the Dengue
Dengue
result showed virus
thatdetection
the highest results
TII inofDHF
unbloodfed
endemic DHF
virus in
TTI5on villages
Aedes insp Malalayang
mosquitoes district,
with thewhile
IR of
Ae. albopictus samples on the F1
areas was in Malalayang I and Malalayang II with generation in the
DHF in 5 villages in Malalayang district, whiler
value of closeness of the relationship with
5 DHF
TII 20%,endemic
whereas, villages
the the can be seen
lowest in Table
TII was in East5, =the
0.38 indicates
value the correlation
of closeness was not strong.
of the relationship with r
whereas theI. differences between Ae. aegypti
Malalayang This is possible because of the ease
= 0.38 indicates the correlation was not strong.transport
andDengue
Ae. albopictus Dengue results
virus detection virus TTI of can be seen
unbloodfed between regionbecause
This is possible led to ofincrease
the easepopulation
transport
in
Ae.the Table 6, and
albopictus the differences
samples between Ae.
on the F1 generation in mobility,
between allowing
region theled spread of dengue
to increase viruses
population
aegypti and Ae. albopictus
5 DHF endemic villages can Dengue
be seen virus positive
in Table 5, from other
mobility, regions.
allowing theInspread
this study, patients
of dengue with
viruses
rate can be seen in the Table 7.
whereas the differences between Ae. aegypti DHF case data were not classified by
from other regions. In this study, patients with patient
andTable 5 showed that
Ae. albopictus Denguetransovarial
virus TTItransmission
can be seen age,
DHF so there
case datawaswere
the posibility of cases
not classified by derived
patient
of dengue virus in Ae. albopictusonly
in the Table 6, and the differences between occuredAe. in from other areas because of the high population
age, so there was the posibility of cases derived
Batukota
aegypti and with
Ae.TTI of 4.5%.
albopictus The result
Dengue showed
virus positive mobility.
from other In areas
contrast study of
because result
the done by Sucipto
high population
that TTI be
rate can of seen
Ae. aegypti was significantly
in the Table 7. higher (2009),
mobility.patient casestudy
In contrast limited ondone
result children aged
by Sucipto
thanTable
Ae. albopictus.
5 showed that Thetransovarial
results were similiar to
transmission under
(2009),five, assuming
patient the caseoncomes
case limited fromaged
children the
those
of dengueperformed byAe.
virus in Wanti (2010) inonly
albopictus Kupang, that
occured local
undersite
five,. assuming the case comes from the
12

found
in Batukotathat with
Ae. aegypti TTI (20.14%)
TTI of 4.5%. The result wasshowed
higher local site12.
than
that TTIAe. albopictus
of Ae. aegypti(8.33%)
was .significantly higher
11
CONCLUSION AND
thanThe Ae.result of Fisher`s
albopictus. Exact were
The results Test confirmed
similiar to SUGGESTION
that the positive rates of Ae.
those performed by Wanti (2010) in Kupang, that aegypti was CONCLUSION
significantly
found higher
that Ae. than TTI
aegypti Ae. (20.14%)
albopictus (Table
was 7).
higher This study found the existence of Dengue
BasedAe.
than onalbopictus
the results(8.33%)
of microscopic
11
. examination virus transovarial
This study found withthedifferent TTI inofeach
existence DHF
Dengue
of theThe Aedessp
result ofmosquito,
Fisher`s Exact the Test highest TTI in
confirmed endemic villages with
virus transovarial ranged from TTI
different 6.1%in to
each17.1%.
DHF
DHF
that endemic areasrates
the positive was inof Malalayang
Ae. aegyptiII with was The Dengue virus transovarial
endemic villages ranged from 6.1% to 17.1%. transmission
TII 17.1%, followed by Malalayang I
significantly higher than Ae. albopictus (Table 7). 13.3%, Bahu was
The different
Dengue virus between Ae. aegypti
transovarial and Ae.
transmission
10%,
Based Matukota
on the results7.7% and East Malalayang
of microscopic examinationI albopictus,
was withbetween
different TTI in Ae.Ae. albopictus
aegyptiwasandlower
Ae.
village
of the 6.1%Aedes(Table 8).
sp mosquito, the highest TTI in than Ae. aegypti’s.
albopictus, with TTI Statistic test result
in Ae. albopictus wasshowed
lower
DHFToendemic
assess the
areasrelationship between IIAedes
was in Malalayang with no
thansignificant correlation
Ae. aegypti’s. between
Statistic the Aedessp
test result showed
spp with DHF incidence rate
TII 17.1%, followed by Malalayang I 13.3%, Bahu (IR), Pearson mosquito Dengue virus TTI with DHF
no significant correlation between the Aedessp IR in 5 DHF
correlation
10%, Matukota statistic test East
7.7% and wasMalalayang
done and p-valueI village endemic
mosquitovillages
DengueinvirusMalalayang
TTI with district.
DHF IR inFurther
5 DHF
=6.1%
0.528 was obtained.
(Table 8). It indicates that there was research needs to be done with more
endemic villages in Malalayang district. Further number of
no significant
To assess the correlation between
relationship between the Dengue
Aedes villages,
researchinneeds
ordertotobeget complete
done data number
with more from each of
virusTTIon Aedes spp mosquitoes
spp with DHF incidence rate (IR), Pearson with the IR of endemic and sporadic DHF villages.
villages, in order to get complete data from each
correlation statistic test was done and p-value = endemic and sporadic DHF villages.
0.528 was obtained. It indicates that there was

100 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.