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The red weaver ant, Oecophylla smaragdina as vectors of bacteria in urban

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International Journal of Entomology Research

International Journal of Entomology Research

ISSN: 2455-4758; Impact Factor: RJIF 5.24
Received: 17-11-2018; Accepted: 19-12-2018
www.entomologyjournals.com
Volume 4; Issue 1; January 2019; Page No. 46-49

The red weaver ant, Oecophylla smaragdina as vectors of bacteria in urban environments
Thangavel Rajagopal1*, Pitchai Singam2, Subbiah Kulandaivel3, Subbiah Selvarani4,
Subramanian Sevarkodiyone5, Ponnirul Ponmanickam6
1-4
Post Graduate and Research Department of Zoology and Microbiology, Thiagarajar College (Autonomous),
Madurai, Tamil Nadu, India
5, 6
Department of Zoology, Ayya Nadar Janaki Ammal College (Autonomous), Sivakasi, Tamil Nadu, India

Abstract
The present study aims to isolate and characterize bacteria from the gut of the workers of the red weaver ant, Oecophylla
smaragdina. The red weaver ants collected from urban and rural areas located in and around Madurai City. The gut contents
were extracted and streaked on nutrient agar medium. The predominant types of bacterial colonies were isolated and streaked to
get pure colonies and were subjected to morphological and biochemical tests for identification. Three bacterial isolates (RH1-
Bacillus sp.1, RH2-Bacillus sp.2 and RH3-Bacillus sp.3) from rural and three (UH1-Bacillus sp.1, UH2-Klebsiella sp. (UH2)
and UH3-Bacillus sp.2.) from urban habitats were identified based on their morphological and biochemical features. The
Klebsiella spp., a pathogenic bacteria carried by the red weaver ant, Oecophylla smaragdina in its gut could be a source of
infections in the urban environment.

Keywords: Oecophylla smaragdina, urban habitat, gut microbes, Tamil Nadu, Klebsiella spp.

1. Introduction isolation and characterization of the gut community have

Ants (family-Formicidae) play an integral role in terrestrial
ecosystems such as aerate soil, disperse seeds, and serve as a
food source for other organisms. Among insect’s ants, the
Oecophylla is prominent member of the forest insect
communities’ throughout the entire tropical world [1, 2].
Several reports on the characterization of microbial
communities associated with 62 different insect species
including ants from various orders highlighted that the
microbial composition of distantly related insect taxa [3].
Symbiotic microbes are ubiquitously located in insect guts
with these symbioses ranging from pathogenic to mutualistic
and from facultative to obligate [4]. The diversification and
evolutionary success of insects have associations with many
microorganisms, most of the gut microbes play a significant
role to their hosts such as in aid digestion of recalcitrant food
components, providing the nutrients directly to their hosts,
assisting in detoxifying ingested particles, protect from
predators, parasites, and pathogens; contribute to inter- and
intraspecific communication; affect efficiency as disease
vectors etc., [5, 6]. For instance, Buchnera spp. are the
symbionts of pea aphid, they providing the nutritional
supplements such as amino acids and vitamins to the pea
aphid [7], carpenter ants (genus Camponotus), have the gut
bacteria in the genus Blochmannia, that provide nutrition to
the host [8], Wolbachia species to provide vitamin C and
antiviral agent to protect the Drosophila melanogaster
against viral infection [9], Leucocoprinus species associated
with leaf cutter ants, can produce antibacterial agents that
help to defend their fungal gardens [10] etc.
It is reported that the microorganisms associated with the
insect may have an important role for human infectious
diseases, that microbes may be modified in order to involve
the pathogen development and, consequently, the disease
transmission [11, 12]. The red weaver ants are prime candidates
to examine the microbial symbiotic interactions. To date, the
largely been studied in insects and limited to ant’s species [13].
However, in- vitro studies of gut microbita have not yet been
carried out in the Oecophylla smaragdina. Therefore, the
present study identified the gut bacteria isolated from red
weaver ants captured in urban and rural areas. Further, we
also identify this ant’s ability to carry any human pathogenic
microbes in its environment.
2. Materials and Methods
Collection of red weaver ant and preparation of
homogenates
The red weaver ants collected from the two sites were located
in the Madurai District, Tamil Nadu, they are broadly into
two categories reflecting different levels of disturbance,
namely rural habitats in agricultural area (relatively less
disturbed) and urban habitats in residential area (relatively
more disturbed). Selection of study sites in this manner
ensured that these sites represented habitats under different
environmental conditions and levels of disturbance.
The Oecophylla smaragdina worker ants were taken out of
their nests from selected localities of urban and rural areas
and placed in sterilized petriplates separately. They were then
surface sterilized by washing with 70% alcohol and then ants
were degutted using fine tipped forceps and were crushed
with the help of glass rods. The paste obtained from the ants
gut was used for isolation of the bacteria with the help of
inoculating loops.
Isolation and screening of bacteria from the ant gut
1g of each ant gut paste (urban and rural ant gut) was taken
and mixed in 99 ml of sterile distilled water. 1ml from each
sample was taken and serially diluted in 9 ml of sterile
distilled water up to 10-9 and plated on the nutrient agar plates
separately and incubated at 37oC for 24 hours for colony
formation. After the incubation period, the plates were

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International Journal of Entomology Research

examined and bacterial colonies were selected. The colonies
were then purified by subculture on plates.
Characterization of bacterial isolate
Bacterial isolates were examined and identified based on their
morphological [cell morphology, endospore formation and
mobility] and biochemical properties [gram staining, oxidase,
catalase, citrate utilization, hydrogen sulphide, methyl red
reaction, voges-proskauer, indole production, starch
hydrolysis (amylase), casein hydrolysis (protease) tests].
3. Results and Discussion
Several studies have been carried out to analyze the microbial
diversity associated with ants [14, 15]. However, the symbiotic
relationships of ants with microbes have rarely been studied
[16]
. Total Six bacterial isolates were isolated from the gut of
red weaver ant using nutrient agar medium, the isolates RH1,
RH2 and RH3 were isolated in worker ants from the rural
habitats, while UH1, UH2 and UH3 isolates from urban
habitats. The colonies of the two isolates (RH3 and UH3) had
large smooth texture, irregular, motile and flat elevation,
whereas four isolates (RH1, RH2, UH1 and UH2) had white
milky texture, rod shape, motile and flat elevation. Based on
the Bergys manual and biologic reader, isolates RH1, RH2,
RH3, UH1 and UH3 were identified as Bacillus spp., this
kind of bacteria has been identified several invertebrates
species and form a significant portion of the intestinal
microbial community. For example, in termites, the Bacillus
spp. play a role in the digestion of polysaccharides and
aromatic compounds [17], in earthworms, they responsible for
the production of nitrous oxide [18], in collembola, they
degrading the chitin [19], in millipedes [20] and in P. scaber
(Isopoda), they responsible for the cellulose degraders [21].
Bacillus species have been found in the hemolymph of fire
ants [22] and gut of Holotrichia parallela larvae [23]. The result
suggests that the Bacillus spp. which may be able to provide
the nutritional supplements and may engage digestion of
biological materials in the gut of red weaver ants.
We observed Klebsiella (isolate UH2) species in gut of red
weaver ants from urban habitats and were not found in the
rural habitats. Klebsiella belong to the Proteobacteria family
Enterobacteriaceae, which occur widely in the guts of
Lepidoptera insects, herbivores and and humans, as noted,
this species presents great heterogeneity in its ecology, host
range, and pathogenic potential [24, 25]. Maximo et al. [26] found
that 4.2 % of Klebsiella spp. was identified in the gut of
different species of ants such as Paratrechina, Monomorium,
Dorymyrmex, Pheidole and Brachymyrmex. Klebsiella
bacterial species also were screened in red imported fire ant
[27, 28]
. It is reported that the Klebsiella pneumoniae is a
pathogen common cause of nosocomial infections that
include urinary tract, respiratory, and wound infections [29, 30].
Cassettari et al. [31] reported that the Klebsiella spp. was
responsible for a 5.6% rate of urinary tract infections,
conjunctivitis and bacteraemia in a public hospital in São
Paulo, Brazil. Klebsiella spp. has been isolated from ants in
hospital environments and 35% of infections were caused by
Klebsiella spp. in the hospital environment [32]. Nagao et al.
[33]
reported that the Klebsiella species is an important
causative agent of nosocomial infection and primary
bloodstream infection.
The results show that the Oecophylla smaragdina ant in both
habitats that can be infected with Gram-positive and Gram-
negative bacteria. In urban habitats, the ants may carry the
human pathogenic bacteria i.e. Klebsiella spp. (isolate UH2),
which highlights their remarkable spreading and transmitting
the diseases in the precise environment. The present study
suggests the red weaver ant may be one of agent (i.e., vector)
responsible for the transmission of disease in the urban
environment.

Table 1: Morphological and Biochemical test for the identification of bacterial isolate from the gut of red weaver ant
Rural Habitat Urban Habitat
S. No. Test
RH1 RH2 RH3 UH1 UH2 UH3
A: Morphological Study
1 Gram staining + + + + - +
2 Shape Rod Rod Rod Rod Rod Rod
3 Motility Motile Motile Motile Motile Motile Motile
4 Cultural characteristics Milky White Milky White Large, Smooth Milky White Milky White Large, Smooth
B: Biochemical Test
5 Indole production test + + - + - -
6 Methyl red reaction (MR) + + + + - +
7 Voges - Proskauer test (VP) + + + + + +
8 Citrate utilization - - + - + +
9 Oxidase + + + + + +
10 Catalase + + + + - +
11 H2S production + + - + + -
12 Amylase + + + + + +
13 Protease + + + + + +
Organism Bacillus sp. 1 Bacillus sp. 2 Bacillus sp. 3 Bacillus sp. 1 Klebsiella sp. Bacillus sp. 2

4. Acknowledgements
The authors give thanks the Principal and the Management of
Thiagarajar College (Autonomous), Madurai for providing
the facilities and the constant encouragement. TR and PP
acknowledges the Science and Engineering Research Board
(SERB) for the award of Fast -Track Young Scientist
Fellowship, New Delhi.
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International Journal of Entomology Research
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