Acta Tropica 153 (2016) 5763

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Acta Tropica
journal homepage: www.elsevier.com/locate/actatropica

Breeding habitat preference of preimaginal black ies (Diptera:

Simuliidae) in Peninsular Malaysia
Zubaidah Yacob a, , Hiroyuki Takaoka a , Pairot Pramual b , Van Lun Low a ,
Mohd Soan-Azirun a
a
b

Institute of Biological Sciences, Faculty of Science, University Malaya, 50603 Kuala Lumpur, Malaysia
Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham 44150, Thailand

a r t i c l e

i n f o

Article history:
Received 22 June 2015
Received in revised form 7 October 2015
Accepted 10 October 2015
Available online 18 October 2015
Keywords:
Simuliidae
Simulium
Ecology
Habitat characteristics

a b s t r a c t
To investigate the breeding habitat preference of black ies, a comprehensive black y survey was conducted for the rst time in Peninsular Malaysia. Preimaginal black ies (pupae and larvae) were collected
manually from 180 stream points encompassing northern, southern, central and east coast of the Peninsular Malaysia. A total of 47 black y species were recorded in this study. The predominant species
were Simulium trangense (36.7%) and Simulium angulistylum (33.3%). Relatively common species were
Simulium cheongi (29.4%), Simulium tani (25.6%), Simulium nobile (16.2%), Simulium sheilae (14.5%) and
Simulium bishopi (10.6%). Principal Component Analysis (PCA) of all stream variables revealed four PCs
that accounted for 69.3% of the total intersite variance. Regression analysis revealed that high species
richness is associated with larger, deeper, faster and higher discharge streams with larger streambed
particles, more riparian vegetation and low pH (F = 22.7, d.f. = 1, 173; P < 0.001). Relationship between
species occurrence of seven common species (present in >10% of the sampling sites) was assessed. Forward logistic regression analysis indicated that four species were signicantly related to the stream
variables. S. nobile and S. tani prefer large, fast owing streams with higher pH, large streambed particles
and riparian trees. S. bishopi was commonly found at high elevation with cooler stream, low conductivity, higher conductivity and more riparian trees. In contrast, S. sheilae was negatively correlated with
PC-2, thus, this species commonly found at low elevation, warmer stream with low conductivity and
less riparian trees. The results of this study are consistent with previous studies from other geographic
regions, which indicated that both physical and chemical stream conditions are the key factors for black
y ecology.
2015 Elsevier B.V. All rights reserved.

1. Introduction
Adult black ies (Diptera: Simuliidae) are one of the most important groups of blood-sucking Diptera (Takaoka, 1983). The females
of certain species, when they bite and draw blood, not only cause
severe skin diseases to humans and animals, but also serve as a vector of larial parasites (Ishii et al., 2008). The Simuliidae is widely
distributed across all zoogeographical regions and the preimaginal
black ies generally inhabit unpolluted running water (Takaoka,
1995; Currie and Adler, 2008). In fact, preimaginal black ies are
important components of the stream ecosystem (Hamada et al.,
2002; Currie and Adler, 2008; Pramual and Kuvangkadilok, 2009).
They act as the keystone species in the ecology of running water,

Corresponding author.
E-mail address: xuehy perdana@yahoo.com (Z. Yacob).
http://dx.doi.org/10.1016/j.actatropica.2015.10.007
0001-706X/ 2015 Elsevier B.V. All rights reserved.

because they are usually present as a major component of stream

macroinvertebrates (Malmqvist et al., 2004) and have an ability to
lter dissolved organic matter and make it available in the food
chain (Currie and Adler, 2008). Black ies are also important in
the monitoring of freshwater contamination because larvae and
pupae are susceptible to both organic and inorganic pollution (i.e.,
insecticides and fertilizers) (Currie and Adler, 2008).
Although black ies are well known for their medical and ecological importance, the signicance of Malaysian black ies remains
unexplored. By contrast, the spatial distribution of preimaginal
black ies has been well-studied in many parts of the world, for
example in the temperate region (McCreadie and Adler, 1998,
2006; McCreadie et al., 2005), the tropical region of South America (Grillet and Barrera, 1997; Hamada and McCreadie, 1999;
Hamada et al., 2002; McCreadie et al., 2004) and the Oriental region,
notably Thailand (Pramual and Kuvangkadilok, 2009; Pramual
and Wongpakam, 2010). These previous studies reported that the

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Z. Yacob et al. / Acta Tropica 153 (2016) 5763

species richness and distribution of preimaginal black ies have

been primarily associated with stream physicochemical conditions.
Ecological studies of black ies in the Oriental region, particularly South East Asia are still scarce except in Thailand, where the
advances in simuliid morphotaxonomy (Takaoka and Choochote
2004), cytotaxonomy (Phasuk et al., 2005; Kuvangkadilok et al.,
2008; Tangkawanit et al., 2009; Pramual et al., 2012; Pramual and
Adler, 2014) and phylogenetics (Pramual et al., 2011; Pramual and
Adler, 2014) have allowed researchers to gain a better understanding of their ecology (Pramual and Kuvangkadilok, 2009; Pramual
and Wongpakam, 2010). Many countries in Southeast Asia are
experiencing extremely slow growth in the taxonomic knowledge
of black ies, as a consequence, nothing is known about their
ecology, biology and other related information. In Malaysia, morphotaxonomic studies of black ies have been progressing well
in recent years, (Takaoka and Davies, 1995; Takaoka, 2012) thus,
allowing us to make our rst attempt to obtain insight into the
black y ecology in association with their habitat characteristics.
Exploring the fauna of black ies is a prerequisite for larval habitat
control programs. Hence, the current study aims to investigate the
breeding habitat preference of preimaginal black ies for the rst
time, across four geographical regions in Peninsular Malaysia.
2. Material and method
2.1. Study sites
This study was carried out at 180 stream points across Peninsular Malaysia, encompassing four geographical regions and 10 states
namely East coast (Kelantan, Terengganu and Pahang), Northern
(Perlis, Kedah, Penang and Perak), Central (Selangor) and Southern
(Negeri Sembilan and Johore) (Fig. 1). The sampling sites included
forests, recreational areas, agricultural sites (oil palm and rubber
plantations), and residential areas. Details on sampling sites and
collections are presented in Table 1.
2.2. Preimaginal sampling and identication
Samplings were conducted within March 2013 until February
2015 (Table 1). There is no distinct wet or dry season throughout
the year and rain is experienced every single month in Malaysia
with an average 2000 to 3500 mm per year. Seasonal rainfall variation occurred in every state of Malaysia during the northeast and
northwest monsoon seasons (Low et al., 2012). We ensured that
our sampling periods were free from its inuence. All potential
breeding sites for black ies were chosen based on accessibility
and the presence of ow (i.e., streams, waterfalls and ditches). Each
stream was sampled once from downstream to upstream (30 m), for
approximately 1 h, by two people. Larvae and pupae attached on
aquatic substrates such as grasses, leaves and stems, twigs, plant
roots and rocks were collected by hand using ne forceps. These
sampling protocols could represent the species occurrence in a
locality (McCreadie and Colbo, 1991; McCreadie et al., 2005). Pupae
attached on similar substrates were individually kept alive in vials
until emergence. The adults, together with their pupal exuviae and
cocoons were preserved in 80% ethanol for identication at the subgenus, species-group or species level. The methods of collection
and identication followed those of Takaoka (2003) and Adler et al.
(2004).
2.3. Physicochemical measurement
The following stream physicochemical parameters were measured at the time of each collection: width (m), depth (m), velocity
(m/s) (one to three measurements along the collection path), temperature ( C), acidity (pH), conductivity (mS/cm) and dissolved

oxygen (mg/L). The values of pH, temperature, conductivity and

dissolved oxygen were taken using a portable multi probe parameter (Hanna HI 9828). Meter tape and steel ruler were used to
measure stream width and depth, respectively, while a cork and
a timer watch were used to measure stream velocity; the time
taken for a cork to move one meter in distance. Velocity, depth and
width measurements were used to estimate discharge (McCreadie
et al., 2006). The physicochemical measurements protocols including those for major streambed particles, riparian vegetation, and
canopy cover followed those of McCreadie et al. (2006). For each
study site, the latitude and longitudinal co-ordinates were taken
and recorded using a hand held Global positioning system (GPS)
instrument (Garmin International Inc., Olathe, KS).
2.4. Data analysis
The frequency of species occurrence was designated in percentages. The presence or absence of a species was expressed
on a binary scale (0 = absent, 1 = present), as in previous studies (Hamada and McCreadie, 1999; McCreadie et al., 2004 and
Pramual and Kuvangkadilok, 2009). Because stream variables are
inter-correlated, Principal Components Analysis (PCA) was used to
reduce the number of variables into groups of independent components. Principal Components (PCs) with eigenvalues greater than
1.0 were retained as variables. To interpret the PCs, Spearmans rank
correlations were used to detect the relationship between principal components and stream variables using a signicance level of
P < 0.001. Forward logistic regression analysis was used to examine
the relationships between spatial distribution and the PCs. Only
species that occurred at more than 10% of the sites were considered in regression analyses (Hamada et al., 2002) because those
present at a lower frequency have resulted in the lack of statistical power (large number of zero values were observed) (McCreadie
et al., 2005). Linear regression was used to test the relationship
between species richness (i.e., number of species in each sampling
site) and the stream variables of the sampling sites (i.e., PC scores).
All collections (180 stream points) were subjected to PCA, and the
PC scores were used for regression analysis.
3. Result
Forty-seven species were found from 180 sampled streams
throughout Peninsular Malaysia (Table 2). At the subgeneric level,
Gomphostilbia (31 species) was the largest subgenus in our collection, followed by Simulium s. str. (12 species) and Nevermannia
(4 species). The current study also successfully revealed 16 out of
the 18 species-groups recorded in Malaysia. The Simulium batoense
species-group (9 species) was the most abundant, followed by the S.
asakoae species-group (7 species), the S. ceylonicum species-group
and the S. epistum species-group (4 species each). Other speciesgroups were represented by one to three species.
The predominant species were S. trangense (36.7%) and S.
angulistylum (33.3%). Relatively common species were S. cheongi
(29.4%), S. tani (25.6%), S. nobile (16.2%), S. sheilae (14.5%) and S.
bishopi (10.6%). Other species were collected at frequencies lower
than 10% (Table 2). Based on the current results, 85.1% (40 species)
of total collected species had frequency of occurrence less than 10%
and were considered as rare (Table 2). Of these, 12 species were collected only once (0.6%) (Table 2). The maximum number of black y
species collected in a single stream for all samples (180 sampling
points) was 10 and minimum was one with the mean number per
stream for the total sample sites was 3.0 0.1 (SE).
The PCA of all collections (180 sampling points) revealed four
PCs which had eigenvalues >1.0, accounted for 69.3% of the total
intersite variance of the stream condition variables (Table 3). Spear-

Z. Yacob et al. / Acta Tropica 153 (2016) 5763

59

Fig. 1. Map showing the location of sampling points in Peninsular Malaysia, small maps showing the Southeast Asia countries (top right) and Langkawi Island in the state of
Kedah (bottom left).

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Z. Yacob et al. / Acta Tropica 153 (2016) 5763

Table 1
Local and regional richness of black ies in Peninsular Malaysia.
Region
Northern
Perak
Batang Padang
Bukit Larut
Pulau Pangkor
Lenggong
Kinta
Kuala Kangsar
Kedah
Langkawi
Kulim
Kuala Muda
Yan
Padang Terap
Baling
Perlis
Kaki Bukit
Mata Ayer
Penang
Balik Pulau
Southern
Johore
Ledang
Kota Tinggi
Pontian
Mersing
Negeri Sembilan
Jelebu
Central
Selangor
Hulu Langat
HuluSelangor
East coast
Kelantan
Gua Musang
Jeli
Pasir Puteh
Kuala Krai
Tanah Merah
Terengganu
Dungun
Hulu Terengganu
Pahang
Jerantut
Lipis-Merapoh
Raub
Cameron Highland

GPS coordinates

Collecting date

Total collections

Regional richness

Mean local richness (SE)

04.2404.58 N 101.71101.72 E
04.8604.87 N 100.76100.89 E
04.2404.25 N 100.56100.57 E
05.1705.18 N 100.93101.10 E
04.3504.71 N 100.07101.24 E
04.6404.83 N 100.85100.87 E

February-15
February-15
February-15
February-15
February-15
February-15

March-13
March-13

21
5
2
2
4
5
3
30
17
6
2
1
1
3
5
4
1

23
16
4
3
8
9
9
20
10
8
3
6
4
7
5
5
1

3.95 0.39
6.25 0.48
2.00 1.00
2.00 0.00
3.50 0.29
3.40 0.55
4.33 0.88
2.57 0.23
2.06 0.22
2.80 0.49
2.50 0.50

3.00 0.00
1.81 0.37
2.00 0.41

06.3706.42 N 099.67099.87 E
05.4005.41 N 101.76101.78 E
05.4105.81 N 100.44101.43 E
05.79 N 100.40 E
06.37 N 100.56 E
05.7205.99 N 100.81100.90 E

March-13
March-13
March-13
Mar-13
March-13
March-13

06.4206.98 N100.02100.11 E
06.55 N 100.17 E
05.0105.57 N 100.09100.98 E

December-14

2.00 0.58

02.0702.36 N 102.53102.64 E
02.3702.83 N 102.61103.87 E
01.3701.83 N 103.51103.87 E
01.81 N 103.87 E

March-14
March-14
March-14
March-14

16
9
3
3
1

16
12
8
4
1

2.81 0.27
2.56 0.34
3.67 0.67
3.33 0.33

02.8303.02 N 102.03102.04 E

January-14

2.67 0.33

03.2103.89 N 101.41102.31 E
03.3303.67 N 101.68101.75 E

April-15
April-15

12
4
8

15
7
14

3.42 0.70
2.25 0.95
4.00 0.90

04.6804.73 N 101.52101.68 E
05.0905.11 N 101.50101.56 E
05.74 N 102.37 E
05.31 N 102.29 E
05.1205.14 N 101.51101.57 E

April-13
Feb-15
April-13
April-13
February-15

04.5404.59 N 102.94102.96 E
04.7704.97 N 102.75102.84 E

August-13
August-13

03.0204.93 N 102.12102.89 E
04.4004.65 N 102.14102.40 E
03.4503.72 N 101.71101.72 E
04.4004.58 N 101.34101.38 E

May and April 2014

September-13
April-15
September-14

21
7
4
1
1
8
12
7
5
56
30
19
4
3

18
7
11
5
4
12
13
8
8
37
19
11
13
11

3.81 0.55
2.00 0.49
5.75 1.60

4.25 0.88
2.83 0.32
2.86 0.50
2.80 0.34
2.67 0.22
2.67 0.34
2.53 0.28
3.75 1.49
3.00 1.00

mans rank correlations revealed that sites with a higher PC-1,

which explained 28.4% of the total variance, were wider, deeper,
faster and had low pH, higher discharge, larger streambed particle
size, with more riparian vegetation. PC-2 explained 18.3% of the
total variance. Sites with a higher PC-2 score were at higher altitudes, had cooler stream temperatures, low conductivity, higher
dissolved oxygen, with more canopy cover and riparian trees. PC-3
accounted for 12% of the total variance. Sites with a higher PC3 score had higher dissolved oxygen and pH with more stream
cover. PC-4 accounted for 10.6% of the total variance. Sites with
a higher PC-4 were at higher altitudes, had cooler water, and
larger streambed particles. This PC explains similar stream variables to PC-2 because both signicantly related to altitude and
water temperature. However, PC-2 was also signicantly related
to water conductivity and dissolved oxygen while these variables
were not related to PC-4. Similarly, PC-4 was signicantly related
to streambed particle while PC-2 did not (Table 3). Therefore,
these two principal components were explained in different stream
variable. Regression analysis between species richness and PCs
indicated that the species richness was positively and signicantly
associated with PC-1 (F = 22.7, d.f. = 1, 173; P < 0.001). Therefore,

more species were found in streams which were larger, deeper, and
owing faster with higher discharge, larger streambed particles,
more riparian vegetation and high pH.
Forward logistic regression analysis was conducted for seven
species (S. trangense, S. angulistylum, S. cheongi, S. bishopi, S. nobile,
S. tani and S. sheilae) which were found in more than 10% of the
sampling sites (Table 2). All of the regression models of species distribution, except S. trangense, S. angulistylum and S. cheongi, were
signicant at P < 0.001 with correct classication varying from 76
to 86.1% (Table 3). PC-1 was signicantly correlated with the occurrence of three species (S. sheilae, S. tani and S. nobile). Occurrence
of S. sheilae was negatively correlated with PC-2, and in contrast, S.
bishopi was positively correlated with PC-2. The occurrences of S.
tani and S. nobile was positively associated with PC-1. The PC-3 and
PC-4 were not signicantly related to any species.
4. Discussion
This comprehensive black y survey was conducted for the rst
time in Peninsular Malaysia and yielded 75.8% of current total
black y species inhabiting Peninsular Malaysia (62 species) (Adler

Z. Yacob et al. / Acta Tropica 153 (2016) 5763

61

Table 2
Frequency of occurrence (FO) for 47 black y species recorded from 180 sampling points across Peninsular Malaysia.
Species

FO

Simulium (Gomphostilbia) adleri Jitklang and Kuvangkadilok (2008)

Simulium (Gomphostilbia) angulistylum Takaoka and Davies (1995)
Simulium (Gomphostilbia) asakoae Takaoka and Davies (1995)
Simulium (Gomphostilbia) azhari Takoaka et al. (2014)
Simulium (Gomphostilbia) aziruni Takaoka et al. (2012)
Simulium (Gomphostilbia) brinchangense Takaoka et al. (2014)
Simulium (Gomphostilbia) burtoni Takoaka and Davies (1995)
Simulium (Gomphostilbia) charlesi Takaoka (2008)
Simulium (Gomphostilbia) cheongi Takaoka and Davies (1995)
Simulium (Gomphostilbia) decuplum Takaoka and Davies (1995)
Simulium (Gomphostilbia) duolongum Takaoka and Davies (1995)
Simulium (Gomphostilbia) gombakense Takaoka and Davies (1995)
Simulium (Gomphostilbia) izuae Takaoka et al. (2013)
Simulium (Gomphostilbia) jerantutense Takaoka et al. (2014)
Simulium (Gomphostilbia) johorense Takaoka et al. (2014)
Simulium (Gomphostilbia) kisapense Takaoka et al. (2012)
Simulium (Gomphostilbia) langkawiense Takaoka et al. (2013)
Simulium (Gomphostilbia) leparense Takaoka et al. (2014)
Simulium (Gomphostilbia) longitruncum Takaoka and Davies (1995)
Simulium (Gomphostilbia) lurauense Takaoka et al. (2011)
Simulium (Gomphostilbia) roslihashimi Takaoka and Soan-Azirun (2011)
Simulium (Gomphostilbia) sheilae Takaoka and Davies (1995)
Simulium (Gomphostilbia) soani Takoaka and Hashim (2011)
Simulium (Gomphostilbia) sp. (nr. parahiyangum)*
Simulium (Gomphostilbia) sp. (nr. pegalanense)*
Simulium (Gomphostilbia) tahanense Takaoka and Davies (1995)
Simulium (Gomphostilbia) tanahrataense Takaoka et al. (2014)
Simulium (Gomphostilbia) tekamense Takaoka et al. (2014)
Simulium (Gomphostilbia) terengganuense Takoaka et al. (2012)
Simulium (Gomphostilbia) trangense Takaoka and Davies (1995)
Simulium (Gomphostilbia) whartoni Takaoka and Davies (1995)
Simulium (Nevermannia) aureohirtum Brunetti (1911)
Simulium (Nevermannia) caudisclerum Takaoka and Davies (1995)
Simulium (Nevermannia) hackeri Edwards (1934)
Simulium (Nevermannia) kurtaki Takaoka and Davies (1995)
Simulium (Nevermannia) ledangense Yacob et al. (2014)
Simulium (Nevermannia) sp. (nr. feuerborni)*
Simulium (Simulium) bishopi Takaoka and Davies (1995)
Simulium (Simulium) brevipar Takaoka and Davies (1995)
Simulium (Simulium) grossilum Takoaka and Davies (1995)
Simulium (Simulium) hirtinervis Takoaka and Davies (1995)
Simulium (Simulium) jeffreyi Takoaka and Davies (1995)
Simulium (Simulium) malayense Takaoka and Davies (1995)
Simulium (Simulium) nobile Takaoka & Davies (1995)
Simulium (Simulium) sp. (nr. grisescens)*
Simulium (Simulium) tani Takaoka and Davies (1995)
Simulium (Simulium) yongi Takaoka and Davies (1995)
*

% FO

1
60
2
1
1
1
3
2
53
12
9
11
1
2
1
1
5
2
2
6
15
26
3
17
13
16
1
2
4
65
16
12
1
2
1
1
3
19
13
4
7
8
6
29
14
46
1

0.6
33.3
1.1
0.6
0.6
0.6
1.7
1.1
29.4
6.7
5.0
6.1
0.6
1.1
0.6
0.6
2.8
1.1
1.1
3.4
7.8
14.5
1.7
9.4
7.3
8.9
0.6
1.1
2.2
36.7
8.9
6.7
0.6
1.1
0.6
0.6
1.7
10.6
7.3
2.2
3.9
4.5
3.3
16.2
7.8
25.6
0.6

Undetermined species which probably are the new species.

Table 3
Results of PCA Spearmans rank correlation coefcient between stream variables and principal components (PCs) for 180 collections in tropical streams in Peninsular Malaysia.
Variable

Stream site
Min

Altitude (m)
Temperature ( C)
Width (m)
Depth (m)
Velocity (m/s)
Conductivity (mS/cm)
Dissolved Oxygen (mg/L)
pH
Discharge (m3 /s)
Stream-bed particle
Canopy cover
Riparian vegetation
% Variance explained in PCA
Proportion
Cumulative
**

10.0
16.3
0.05
0.01
0.12
0.0115
1.60
4.35
0.0003
Sand
Open
Open

Principle components
Max
1,345
30.9
15
0.70
1.10
1.83
18.4
8.43
3.15
Bedrock
Forest
Complete

Mean
235.4
23.8
2.54
0.14
0.43
0.306
6.77
6.41
0.34
3a
2a
3a

SE

PC-1

17.0
0.13
0.20
0.009
0.01
0.02
0.27
0.05
0.04

0.176
0.052
0.844**
0.787**
0.808**
0.222
0.005
0.292**
0.914**
0.628**
0.128
0.336**
28.4
28.4

PC-2

PC-3

PC-4

0.392
0.402**
0.045
0.137
0.085
0.476**
0.293**
0.157
0.102
0.128
0.700**
0.678**

0.167
0.043
0.185
0.071
0.161
0.225
0.690**
0.709**
0.149
0.140
0.351**
0.062

0.551**
0.315**
0.084
0.031
0.142
0.231
0.030
0.140
0.095
0.633**
0.021
0.102

18.3
46.7

12.0
58.7

10.6
69.3

**

P < 0.001.
Median values given for riparian vegetation (1 = open, 2 = brush and 3 = forest), streambed-particle size (min; 1 = mud/silt and max; 6 = bedrock), and canopy cover.
(1 = open, 2 = partial and 3 = complete). Rankings followed McCreadie et al. (2006).
a

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Z. Yacob et al. / Acta Tropica 153 (2016) 5763

Table 4
Regression analysis for the distribution of preimaginal black y species in Peninsular Malaysia.
Species

Simulium bishopi
Simulium nobile
Simulium tani
Simulium sheilae

Regression coefcient
K

PC-1

PC-2

PC-3

PC-4

2.275
1.927
1.344
1.876

1.030
0.773

0.716

0.551
0.618

and Crosskey, 2015). Our results indicate that all sampled streams
are the natural breeding habitats for black ies, although some of
the sampling points may have been affected by intensive agricultural practices (e.g., in Cameron Highlands). In fact, the black ies
could serve as the bioindicator for environmental quality assessment, where the immature stages have specic tolerance to habitat
disturbance (Docile et al., 2015). A thorough examination of the
association between black y distribution and degree of impacted
habitat would deepen our knowledge on the biology and ecology
of black ies.
Current result revealed more than half of the black y species
(40 species) assemblages in Peninsular Malaysia are rare species
(FO < 10%). High number of rare species also recorded in most of
the black y studies (Hamada et al., 2002; McCreadie and Adler,
2008; Couceiro et al., 2014). Rare species are important as part
of organism inuencing the function of ecosystem (Lyons et al.,
2005). The species occurrence pattern which largely inuenced
by factors such as species dispersal ability and the stream ecological conditions (Adler et al., 2004; Pramual and Wongpakam,
2010; Couceiro et al., 2014) are the important factors determining whether or not the species is rare. In contrast, 8.5% (four
species) of total collected species (i.e., S. trangense, S. angulistylum, S. tani, and S. cheongi) had frequencies of occurrence ranging
from 25.6 to 36.7% of total sampled streams (Table 2). Black y
species that are widely distributed and adaptable in various physicochemical conditions are likely to be a species complex (Adler and
McCreadie, 1997). This situation has been highlighted in previous
studies where S. tani and S. angulistylum were found as the cytological species complexes (Tangkawanit et al., 2009; Pramual and
Kuvangkadilok, 2012). Our results revealed that this species was
found in a wide range of stream conditions in Peninsular Malaysia.
Therefore, additional cryptic diversity might be found in Malaysian
specimens and further cytogenetic studies would help to clarify this
hypothesis.
Species richness is strongly interconnected with the habitat
characteristics (Hamada et al., 2002). In this study, we found that
species richness was signicantly associated with stream size,
ow, streambed particles, and riparian vegetation. These results
are largely consistent with the pattern of species richness found
in previous studies: large, fast owing streams, large streambed
particles, and cool water at high altitude with cover, and riparian forest along the stream (Pramual and Kuvangkadilok, 2009);
large, fast-owing, high discharge streams, with lower temperature
and conductivity (Pramual and Wongpakam, 2010); larger streams,
cooler water temperature, faster owing, and larger streambed particles (Hamada et al., 2002); larger streams (Grillet and Barrera,
1997); and current velocity and size of streambed particle (Scheder
and Waringer, 2002). Moreover, regression analysis revealed four
common black y species (S. tani, S. nobile, S. sheilae and S. bishopi) are related to stream size, velocity, and riparian vegetation.
Current study revealed the probability of predicting the specieshabitat specic of the most-common species varied from 76% to
89.1% (Table 4). The probability of predicting falls above 70% and
this pattern corroborates with the previous studies (Hamada and
McCreadie, 1999; Hamada et al., 2002; Pramual and Kuvangkadilok,
2009; Pramual and Wongpakam, 2010).

Correct (%)

<0.001
<0.001
<0.001
<0.001

89.1
84.6
76.0
85.1

In addition to the mentioned stream conditions, other associated factors in inuencing black y ecology have also been pointed
out: water chemistry (Townsend et al., 1983; Jenkins et al., 1984;
Erman and Erman, 1995; McCreadie et al., 2005), habitat disturbance (Erman and Erman, 1995; Palmer et al., 1995; Pramual
and Kuvangkadilok, 2009; Couceiro et al., 2014), elevation (Tate
and Heiny, 1995; Srisuka et al., 2015), intense riparian vegetation
(Lautenschlger and Kiel, 2005) as well as the labral fan of black y
larvae (Pangjanda and Pramual, 2015).
In conclusion, this large-scale black y survey has provided
new insight into the black y species composition in Peninsular Malaysia. Current study has revealed that both ecological and
physicochemical are the important factors in inuencing the breeding habitat preference of simuliid species in tropical streams. These
data can be useful for formulating effective prevention and vector control programs (i.e., environmental manipulation and use of
biological control agents based on black y species and density).
Additionally, this rst survey will be the stepping stone promoting more black y studies in Malaysia as well as other countries in
Southeast Asia.

Acknowledgements
Our sincere appreciation goes to the Department of Wildlife
& National Parks (DWNP) and Forestrys Department of Peninsular Malaysia for granting us permission to conduct sampling in all
protected forests. Thanks are due to Nor Azhar Jamil and Muhammad Rasul Abdullah Halim (Institute Biological Sciences, University
of Malaysia) for assistance in the eld. This work was supported
by research grants from the University of Malaya (PG084-2014B
and RP003A-13SUS) and also by the Fundamental Research Grant
Scheme (Project No. FP016-2012A). This project is a part of the rst
authors PhD research at the University of Malaya, Kuala Lumpur.

References
Adler PH, Crosskey RW. 2015. World blackies (Diptera: Simuliidae): A
comprehensive revision of the taxonomic and geographical inventory. 123 pp.
Available from: http://entweb.clemson.edu/biomia/pdfs/blackyinventory.pdf
(accessed 10.09.15.).
Adler, P.H., Currie, D.C., Wood, D.M., 2004. The Black Flies (Simuliidae) of North
America. Cornell University Press. Ithaca, New York, USA.
Adler, P.H., McCreadie, J.W., 1997. The hidden ecology of black ies: sibling species
and ecological scale. Am. Entomol. 43, 153161.
Couceiro, S.R.M., Hamda, N., Sagot, L.B., Pepinelli, M., 2014. Black-y assemblage
distribution in streams in disturbed areas in southern Brazil. Acta Tropica 140,
2633.
Currie, D.C., Adler, P.H., 2008. Global diversity of black ies (Diptera: Simuliidae) in
freshwater. Hydrobiologia 595, 469475.
Docile, T.N., Figueiro, R., Azevedo, H.G., Nessimian, J.L., 2015. Water pollution and
distribution of the black y (Diptera: Simuliidae) in the Atlantic Forest, Brazil.
Revista De Biol. Trop. 63 (3), 683693.
Erman, N.A., Erman, D.C., 1995. Spring permanence: Trichoptera species richness
and the role of drought. J. Kans. Entomol. Soc. 68, 5064.
Grillet, M.E., Barrera, R., 1997. Spatial and temporal abundance, substrate
partitioning and species co-occurrence in a guild of Neotropical blackies
(Diptera: Simuliidae). Hydrobiologia 345, 197208.
Hamada, N., McCreadie, J.W., 1999. Environmental factors associated with the
distribution of Simulium peravum (Diptera: Simuliidae) among streams in
Brazilian Amazonia. Hydrobiologia 397, 7178.

Z. Yacob et al. / Acta Tropica 153 (2016) 5763

Hamada, N.J., McCreadie, J.W., Adler, P.H., 2002. Species richness and spatial
distribution of blackies (Diptera: Simuliidae) in streams of Central Amazonia,
Brazil. Freshwater Biol. 47, 3140.
Ishii, Y., Choochote, W., Bain, O., Fukuda, M., Otsuka, Y., Takaoka, H., 2008. Seasonal
and diurnal biting activities and zoonotic larial infections of two Simulium
species (Diptera: Simuliidae) in northern Thailand. Parasite 15 (2), 121129.
Jenkins, R.A., Wade, K.R., Pugh, E., 1984. Macroinvertebratehabitat relationships
in the River Tei catchment and the signicance to conservation. Freshwater
Biol. 14, 2342.
Kuvangkadilok, C., Lualon, U., Baimai, V., 2008. Cytotaxonomy of Simulium
siamense Takaoka and Suzuki (Diptera: Simuliidae) in Thailand. Genome 51,
972987.
Lautenschlger, M., Kiel, E., 2005. Assessing morphological degradation in running
waters using blacky communities (Diptera, Simuliidae): can habitat quality
be predicted from land use? Limnologica 35, 262273.
Low, V.L., Chen, C.D., Lee, H.L., Lim, P.E., Leong, C.S., Soan-Azirun, M., 2012.
Nationwide distribution of Culex mosquitoes and associated habitat
characteristics at residential areas in Malaysia. Am. Mosquito Control Assoc. 28
(3), 160169.
Lyons, K.G., Brigham, C.A., Traut, B.H., Schwartz, M.W., 2005. Rare species and
ecosystem functioning. Conserv. Biol. 19, 10191024.
Malmqvist, B., Adler, P.H., Kuusela, K., Merritt, R.W., Wotton, R.S., 2004. Black ies
in the boreal biome, key organisms in both terrestrial and aquatic
environment: a review. Ecoscience 11, 187200.
McCreadie, J.W., Adler, P.H., 1998. Scale, time, space, and predictability: species
distributions of preimaginal black ies (Diptera: Simuliidae). Oecologia 114,
7992.
McCreadie, J.W., Adler, P.H., 2006. Ecoregions as predictors of lotic assemblages of
blackies (Diptera: Simuliidae). Ecography 29, 603613.
McCreadie, J.W., Adler, P.H., 2008. Spatial distribution of rare species in lotic
habitats. Insect Conserv. Divers. 1, 127134.
McCreadie, J.W., Hamada, N., Grillet, M.E., 2004. Spatial-temporal distribution of
preimaginal blackies in Neotropical streams. Hydrobiologia 513, 183196.
McCreadie, J.W., Adler, P.H., Grillet, M.E., Hamada, N., 2006. Sampling statistics in
understanding distributions of black y larvae (Diptera: Simuliidae). Acta
Entomol. Serbica Suppl., 8996.
McCreadie, J.W., Adler, P.H., Hamada, N., 2005. Patterns of species richness for
blackies (Diptera: Simuliidae) in the Nearctic and Neotropical regions. Ecol.
Entomol. 30, 201209.
McCreadie, J.W., Colbo, M.H., 1991. Spatial distribution patterns of larval cytotypes
of the Simulium venustum/verecundum complex (Diptera: Simuliidae) on the
Avalon Peninsula, Newfoundland: factors associated with occurrence. Can. J.
Zool. 69, 26512659.
Pangjanda, S., Pramual, P., 2015. Trait-based and phylogenetic community ecology
of black ies (Diptera: Simuliidae) in tropical streams of Thailand.
Hydrobiologia, http://dx.doi.org/10.1007/s10750-015-2389-8.
Phasuk, J., Chanpaisaeng, J., Adler, P.H., Courtney, G.W., 2005. Chromosomal and
morphological taxonomy of larvae of Simulium (Gomphostilbia) (Diptera:
Simuliidae) in Thailand. Zootaxa 1052, 4960.

63

Pramual, P., Adler, P.H., 2014. DNA barcoding of tropical black ies (Diptera:
Simuliidae) of Thailand. Mol. Ecol. Resour. 14, 262271.
Pramual, P., Kuvangkadilok, C., Jitklang, S., Tangkawanit, U., Adler, P.H., 2012.
Geographical versus ecological isolation of closely related black ies (Diptera:
Simuliidae) inferred from phylogeny, geography and ecology. Org. Divers. Evol.
12, 183195.
Pramual, P., Kuvangkadilok, C., 2009. Agricultural land use and black y (Diptera:
Simuliidae) species richness and species assemblages in tropical streams,
Northeastern Thailand. Hydrobiologia 625 (1), 173184.
Pramual, P., Kuvangkadilok, C., 2012. Integrated cytogenetic, ecological, and DNA
barcode study reveals cryptic diversity in Simulium (Gomphostilbia)
angulistylum Takaoka & Suzuki (Diptera: Simuliidae). Genome 55, 447458.
Pramual, P., Wongpakam, K., Adler, P.H., 2011. Cryptic biodiversity and
phylogenetic relationships revealed by DNA barcoding of Oriental black ies in
the subgenus Gomphostilbia (Diptera: Simuliidae). Genome 54, 19.
Pramual, P., Wongpakam, K., 2010. Seasonal variation of black y (Diptera:
Simuliidae) species diversity and community structure in tropical streams of
Thailand. Entomol. Sci. 13, 1728.
Scheder, C., Waringer, J.A., 2002. Distribution patterns and habitat characterization
of Simuliidae (Insecta: Diptera) in a low-order sandstone stream
(Weidlingbach, Lower Austria). Limnologica 32, 236247.
Srisuka, W., Takaoka, H., Otsuka, Y., Fukuda, M., Thongsahuan, S., Taai, K.,
Choochote, W., Saeung, A., 2015. Seasonal biodiversity of black ies (Diptera:
Simuliidae) an devaluation of ecological factors inuencing species distribution
at Doi Pha Hom Pok National Park, Thailand. Acta Trop. 149, 212219.
Takaoka, H., Choochote, W., 2004. A list of and keys to black ies (Diptera:
Simuliidae) in Thailand. Trop. Med. Health 32, 189197.
Takaoka, H., Davies, D.M., 1995. The Black Flies (Diptera: Simuliidae) of West
Malaysia. viii + 175 pp, Kyushu University press, Fukuoka. Japan.
Takaoka, H., 1983. The blackies (Diptera: Simuliidae) of the Philippines. xi + 119
pp. Japan Society For The Promotion of Science, Tokyo, Japan.
Takaoka, H., 1995. The Simuliidae (Diptera) from Bougainville Island, Papua New
Guinea. Jpn. J. Trop. Med. Hyg. 23, 239252.
Takaoka, H., 2003. The Black ies (Diptera: Simuliidae) of Sulawesi, Maluku and
Irian Jaya. xxii + 581 pp. Kyushu University Press, Fukuoka, Japan.
Takaoka, H., 2012. Morphotaxonomic revision of Simulium (Gomphostilbia)
(Diptera: Simuliidae) in the Oriental Region. Zootaxa 3577, 142.
Tangkawanit, U., Kuvangkadilok, C., Baimai, V., Adler, P.H., 2009. Cytosystematics
of the Simulium tuberosum group (Diptera; Simuliidae) in Thailand. Zool. J.
Linn. Soc. 155, 289315.
Tate, C.M., Heiny, J.S., 1995. The ordination of benthic invertebrate communities in
the South Platte River basin in relation to environmental factors. Freshwater
Biol. 33, 439454.
Townsend, C.R., Hildrew, A.G., Francis, J., 1983. Community structure in some
southern English streams: the inuence of physicochemical factors.
Freshwater Biol. 13, 521544.