47th Malaysia Society of Parasitology & Tropical Medicine
47th Malaysia Society of Parasitology & Tropical Medicine
47th Malaysia Society of Parasitology & Tropical Medicine
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~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 47th MSPTM Annual Conference
CONTENTS
Messages
Tan Sri Dato Dr Abu Bakar Suleiman 2
President, International Medical University, Kuala Lumpur, Malaysia
Associate Professor Dr Stephen Ambu 3
President, Malaysian Society of Parasitology and Tropical Medicine
Professor Dr Mak Joon Wah 4
Honorary Member
Dr Reuben Sharma 5
Silver Medal Award
Organising Committee 7
Plenary Speakers 9
Programme
Programme At a Glance 10
Daily Programme 12
List of Posters 19
Abstracts 23
Plenary Session 24
Oral Session 31
Blastocystis Symposia 56
Oral Presentation (Postgraduates) 59
Poster Presentation 74
Acknowledgements 129
Message from
Tan Sri Dato Dr Abu Bakar Suleiman
President, International Medical University, Kuala Lumpur, Malaysia
I am deeply honoured to have been invited by the Malaysian Society of Parasitology and Tropical
Medicine to write this message on the occasion of their 47th Annual Scientific Conference this year.
It is both timely and appropriate that the focus this year is on the theme Climate Change and its
Impact on Public Health. Malaysia has continued to develop rapidly over the last three decades and
this has resulted in many challenges regarding the management of environmental issues that impact
on the peoples wellbeing. As the country progresses, changes to our environmental landscape is
inevitable. Therefore we have to be vigilant and be in the forefront of development to adapt relevant
guidelines and measures to manage the emerging issues for the unimpeded progress of Malaysia
towards its vision of 2020, that is, to achieve a developed nation status.
The impact on human health due to environmental degradation is usually subtle, and the cause, effect
and manifestation of symptoms are realized much later. Therefore adaptation and mitigation measures
must be effective and enforceable. Scientists in Malaysia and the region have contributed much to
the development of appropriate guidelines to address and deal with environmental health problems
appropriately over the years. Nevertheless, there is still need for more research that is current and
relevant to prevailing and emerging environmental conditions and its impact on health.
This conference, I believe, will deliberate on issues that are relevant to the global scenario on
environment and health and bring forth recommendations that will protect public health against the
combined effect of the complexities of modern life and the prevailing environmental pollution.
Message from
Associate Professor Dr Stephen Ambu
President, Malaysian Society of Parasitology and Tropical Medicine
It gives me great pleasure to convey this message on the occasion of the 47th Annual Conference of
the Malaysian Society of Parasitology and Tropical Medicine (MSPTM). The theme of this years
conference is Climate Change and its Impact on Public Health. The conference aims to provide a
platform for scientists from Malaysia and abroad to discuss the various issues relating to environmental
degradation resulting in human sufferings such as increased disease burden and its consequences on
the economy of the affected countries.
The MSPTM has always been in the forefront of biomedical research for the last 47 years, contributing
to the promotion of health, be it human or animal, by addressing current issues for the benefit of
mankind.
In all sincerity I hope that the deliberations at this conference will be of benefit to scientists and policy
planners in the region as well as other countries in Asia, Europe and the Americas.
I would also like to place on record the appreciation of IMU/MSPTM to all those companies which
have made financial contribution for the running of this conference. Our special thanks to Lee
Foundation for their financial donation given in support of the conference. My gratitude is also extended
to MSPTM Council Members, Joint Committee members at IMU, friends and colleagues who have
worked tirelessly to make this conference a success.
Last but not least our appreciation goes to Tan Sri Dato Dr Abu Bakar Suleiman, President, IMU for
mooting the idea of holding a climate change conference at IMU and to Professor Dr Mak Joon Wah
for his support given to make it a reality.
Thank you.
Honorary Member
Professor Dr Mak Joon Wah
Dean of Postgraduate Studies and Research
International Medical University, Kuala Lumpur, Malaysia
Prof Mak Joon Wah, MBBS (Singapore), DAP & E (Malaysia), MPH (Mal), MD (Singapore), FRCPath
(UK), FAMM, FASc
Professor Dr Mak Joon Wah became a member of the MSPTM in 1972 and has since remained an
active member by contributing to the continuous growth of the Society. He has served in the Society
as Council Member 1978 -1980 and 1983; Vice-President 1981; President 1982; Chairman Scientific
Committee Second International Congress of Parasitology and Tropical Medicine. He is the founder/
editor of the MSPTM Journal Tropical Biomedicine which was started in 1985. He continues to be
on the editorial board.
His role in the World Health Organisation has been - Member of the Expert Advisory Panel on Filariasis,
World Health Organization (1981 1997); Member of the Steering Committee of the Scientific Working
Group on Filariasis; and Consultant and Temporary Advisor on many assignments over the years.
Over the years he has been bestowed with various awards such as: Malaysian Society of Parasitology
and Tropical Medicine Medal (1981); National Science Award (Anugerah Sains Negara), Malaysia
(1985); Kesastria Mangku Negara (1986); Sandosham Medal, Malaysian Society of Parasitology and
Tropical Medicine (1989); Excellent Service Award, IMR (1992); Excellent Service Award, Ministry
of Health (19930; Johan Setia Mahkota (1997).
Prof Mak has published over 300 scientific papers and still very actively promotes research among
the younger generation in the areas of bioactive molecules and cellular mechanisms, cancer biology,
pharmaceuticals and drug delivery systems, environmental health research, stem cell research, clinical
research and medical education research at the International Medical University todate.
Reuben Sharma graduated with a Doctor of Veterinary Medicine (DVM) degree from Universiti Putra
Malaysia and subsequently pursued a Master in Veterinary Science (MVSc) in wildlife diseases at
the same university. He was later awarded a PhD in molecular parasitology from the University of
Cambridge, UK. Reuben is also a Chartered Biologist (CBiol) and a Member (MIBiol) of the Institute
of Biology London, and a Fellow of the Royal Society of Tropical Medicine and Hygiene UK.
His main research focus is on the molecular biology, antigenic variation and genetics of parasitic
haemoprotozoa. His research findings has contributed to the revision of the life cycle and ontogeny
of the infective cell stages of Trypanosoma brucei, a representative member of the kinetoplastida
that cause debilitating disease in man and animals. His work has also shed light on chromosome
dynamics, mitotic cell division and gene expression during meiotic recombination in these
haemoprotozoa. Reuben has published over 100 scientific articles in peer-reviewed journals, conference
proceedings and technical reports. He is also keenly interested in morphological and molecular taxonomy
and systematics of Malaysian parasitofauna, wildlife diseases, ecology and conservation. Together with
co-authors, has described 2 new species of parasitic nematodes and redescribed an additional 2 from
chelonians.
Reuben has received 3 international awards for his efforts in the conservation and medicine of
Malaysian wildlife, and has secured additional research grants from the Ministry of Science, Technology
and Innovation (MOSTI), and the Ministry of Higher Education (MOHE) for the study of parasites
and wildlife diseases in Malaysia. He has also received the Excellent Service Award from Universiti
Putra Malaysia.
Over the years Reuben has supervised/co-supervised 17 undergraduate projects and 21 postgraduate
candidates (14 Master and 7 PhD). He has been appointed as consultant/expert resource person to
various NGOs and governmental organizations including World Wide Fund for Nature (WWFM), the
Ministry of Human Resources Malaysia, Ministry of Natural Resources and Environment Malaysia,
and the Veterinary Laboratory Services Unit, UPM. He is also on the editorial board of the Malaysian
Veterinary Journal.
Reuben has served as a council member of the MSPTM for four terms and was the Hon. Secretary
of the 45th MSPTM Council. He is currently a senior lecturer, coordinator of the parasitology laboratory
and coordinator of the research committee at the Faculty of Veterinary Medicine, UPM.
Scientific Committee
Chairperson: Dr Wong Shew Fung
Members : Professor Dr Mak Joon Wah
Professor Dr Chan Boon Tek
Professor Dr Lee Chow Yang
Associate Professor Dr Stephen Ambu
Associate Professor Dr Yvonne Lim Ai Lian
Dr Indra Vythilingam
Dr Nazni Wasi Ahmad
Dr Reuben Sharma
Dr Donald Chen
Dr Chan Li Li
Mr Heo Chong Chin
Mr Chang Kum Wah
Mr Wong Siew Tung
Mr Ooi Soo Shen
Ms Adela Ida Anak Jiram
Secretariat
Ms Danielle Ho
Ms Rosnah Mohd Noor
Ms Yeo Mee Choo
Fund Raising
Associate Professor Dr Stephen Ambu
Mr Wong Siew Tung
Ms Norbazlin Md Marham
Student Competition
Mr Heo Chong Chin
Associate Professor Dr Yvonne Lim Ai Lian
Master of Ceremony
Dr Chew Wai Kit
Supporting Departments
Marketing Department
Facilities Management and Administration Department
E-Learning Department
ITS Department
PLENARY SPEAKERS
Dr Katarzyna Miska
Research Molecular Biologist
Dr Indra Vythilingam Bldg. 1042, BARC-East, 10300 Baltimore Avenue
Principal Research Scientist Beltsville, MD 20705-2350
Environmental Health Institute (EHI) 301/504-5596
National Environment Agency (NEA), Singapore [email protected]
[email protected]
PROGRAMME AT A GLANCE
3rd
MARCH 2011 THURSDAY (DAY 1)
INTERNATIONAL MEDICAL UNIVERSITY
TIME EVENTS
0800 0830 Registration / Breakfast
Atrium / Dewan Cancelor
0830 0845 Arrival of Guests
Auditorium 2
0845 0900 Arrival of Tan Sri Dato Dr Abu Bakar Suleiman
Auditorium 2
0900 0910 Welcome Address
Auditorium 2
0910 1000 OPENING CEREMONY
Auditorium 2
1000 1030 Presentation of
MSPTM Honorary Membership Certificate
MSPTM Silver Medal Award
Auditorium 2
1030 1100 Visit to Exhibition Poster Arena / Tea Break
Dewan Cancelor
1100 1130 Presidential Address
Auditorium 2
1130 1215 PLENARY SESSION 1
Auditorium 2
1215 1315 LUNCH
Dewan Cancelor
1315 1415 PLENARY SESSION 2
Auditorium 2
1415 1515 ORAL SESSION 1 ORAL SESSION 2
Learning Room 4.07
1515 1615 ORAL SESSION 3
Auditorium 2 Learning Room 4.07
1615 1645 TEA BREAK
Dewan Cancelor
1645 1745 PLENARY SESSION 3 BLASTOCYSTIS SYMPOSIA
Auditorium 2 Learning Room 4.07
1745 1945 47th MSPTM AGM MEETING (FOR MEMBERS OF
MSPTM ONLY) Auditorium 2
1945 DINNER
Dewan Cancelor
PROGRAMME AT A GLANCE
4th MARCH 2011 FRIDAY (DAY 2)
INTERNATIONAL MEDICAL UNIVERSITY
TIME EVENTS
DAILY PROGRAMME
3rd
MARCH 2011 THURSDAY (DAY 1)
INTERNATIONAL MEDICAL UNIVERSITY
TIME EVENTS
0900 0910 Welcome Address by the President of MSPTM and Chairman of the
Organising Committee of the 47th Annual Conference of MSPTM
Associate Professor Dr Stephen Ambu
TIME EVENTS
1415 1430 OS1.1: LONGEVITY AND INFECTIVITY OF TRYPANOSOMA EVANSI ISOLATED FROM
THE GUT OF THE STABLE FLY STOMOXYS CALCITRANS (DIPTERA: MUSCIDAE).
GUMAR YS
1515 1530 OS1.5: PARASITIC INFECTIONS IN DOGS AND CATS FROM DOMESTIC AND
HOMELESS ENVIRONMENT. NURHAINIS OGU SALIM
1545 1600 OS1.7: FIELD EVALUATION OF IMR AUTOCIDAL TRAP DEVICE FOR AEDES CONTROL.
NURULHUSNA AH
1600 1615 OS1.8: IMPACT OF PREDATION ON TOXORHYNCHITES SP. FED ON WILD TYPE AND
TRANSGENIC AEDES AEGYPTI (L) LARVAE: IMPLICATION OF RIDL GENE TRANSFER.
OREENAIZA MN
1415 1430 OS2.1: DETECTION OF SCRUB TYPHUS DISEASE USING PCR TECHNIQUE IN CHIGGER
AND TISSUES OF SMALL MAMMALS. AZIMA LH
1430 1445 OS2.2: POINT MUTATIONS IN DHPS AND DHFR GENES OF PLASMODIUM FALCIPARUM
ISOLATES FROM SABAH. NOR AZRINA NORAHMAD
1445 1500 OS2.3: PCR AMPLIFICATION, CLONING AND SEQUENCING OF A GENE ENCODING AN
ERYTHROCYTE INVASION PROTEIN OF PLASMODIUM KNOWLESI. ATIQUE AHMED
1500 1515 OS2.4: OPERATIONAL RESEARCH IN SABAH TO ASSESS THE STATUS OF THE
PROGRAMME FOR ELIMINATION OF LYMPHATIC FILARIASIS. RAHMAH N
TIME EVENTS
1700 1715 BS2: MOLECULAR ASPECTS OF BLASTOCYSTIS SP.: THE ENIGMA CONTINUES.
TAN TC
1715 1730 BS3: BLASTOCYSTIS HOMINIS INFECTION: COULD IT BE LINKED WITH CANCER?
CHANDRAMATHI S
1730 1745 BS4: BLASTOCYSTIS SP. IN WATER SOURCES: CURRENT CHALLENGES AND FUTURE
DIRECTIONS. LEE IL
1745 1945 47th MSPTM AGM MEETING (FOR MEMBERS OF MSPTM ONLY)
1945 DINNER
DAILY PROGRAMME
4th
MARCH 2010 FRIDAY (DAY 2)
INTERNATIONAL MEDICAL UNIVERSITY
TIME EVENTS
0900 0915 OS4.1: DISCOVERY OF A NEW SPECIES OF BLOWFLY FROM CRIME SCENE
INVESTIGATION IN MALAYSIA. KAVITHA RAJAGOPAL
0915 0930 OS4.2: A PRELIMINARY STUDY OF FORENSIC INSECT DIVERSITY IN A HIGH RISE
BUILDING IN SENTUL TIMUR, KUALA LUMPUR. CHEW WK
TIME EVENTS
1045 1100 OP6: GENOTYPING OF TOXOPLASMA GONDII STRAINS ASSOCIATED WITH HUMAN
TOXOPLASMOSIS: A CURRENT STATUS. ANDIAPPAN H
1100 1115 OP7: IMMUNE RESPONSES OF GOATS INFECTED WITH TRYPANOSOMA EVANSI TO
INTRANASAL PNEUMONIC MANNHEIMIA VACCINATION. ABUBAKAR IA
1115 1130 OP8: DEVELOPMENTAL RATE OF SCUTTLE FLY, MEGASELIA SCALARIS (LOEW)
(DIPTERA: PHORIDAE) AT DIFFERENT LABORATORY TEMPERATURES. ZUHA RM
1145 1200 OP10: BLASTOCYSTIS SP.: EVIDENCE OF ITS OCCURRENCE IN WATER SOURCES IN
PENINSULAR MALAYSIA. LEE IL
TIME EVENTS
1515 1530 OS5.4: HIGH LEVEL OF SERUM LIPID DAMAGE IN BREAST CANCER PATIENTS
INFECTED WITH INTESTINAL PARASITES. CHANDRAMATHI S
TIME EVENTS
1600 1630 The Buzzy Business of Genetic Modified Aedes aegypti and Dengue
By Dr Lee Han Lim
Institute for Medical Research, Kuala Lumpur, Malaysia
LIST OF POSTERS
PP11 USING SATELLITE IMAGERY FOR MAPPING MALARIA WAN NAJDAH WMA
TRANSMISSION RISK AREA IN POS LENJANG, PAHANG
PP42 THE DOG LOUSE HETERODOXUS SPINIGER FROM STRAY CATS NORHIDAYU S
IN PENANG, MALAYSIA
PP48 EFFECT OF BEEF LIVER, MEAT AND MIXED NUTRIENT AGAR ZUHA RM
DIETS ON THE DEVELOPMENT OF SCUTTLE FLY, MEGASELIA
SCALARIS (LOEW) (DIPTERA: PHORIDAE)
PP60 STUDY OF FILARIAL PARASITES IN DOGS AND CATS AND THEIR CHEANG SYF
INFECTIONS IN MAN IN SELANGOR
ABSTRACTS
Plenary Session 1
WHO China Office, 401 Dongwai Diplomatic Office Bld 23 Dongzhimenwai Ave Chaoyang Dist,
Beijing, China
China lacks an overarching mitigation and adaptation policy framework to address the health impacts
of climate change. Current barriers include the lack of good quality information and research on the
impacts of climate change on health and the need to develop more effective environmental health
management systems. Effective management of the environment and health interface is critical to
achieving the overall goal of sustainable development to reduce vulnerability and increase the resilience
to climate change impacts. Specific areas to build environmental health capacity include: workforce
development; legislative frameworks; planning, and monitoring and information system development.
This paper provides an overview of the nature, scope and scale of the WHO led, UNDP- Spanish
MDG Program on Climate Change and Health that commenced in 2008. The China National
Environment and Health Action Plan (CNEHAP) forms the framework for the delivery of outputs
related to Climate Change. As such there are four interrelated domains addressed in the Program:
Enhancing EH leadership and management capacities
Implementation of the CNEHAP to local levels
Developing capacity in risk assessment
Developing more effective EH information/monitoring systems.
This paper explores of the process to date, the context within which health issues were integrated
into the Program, the interrelationship between the Program and the China National Environment and
Health Action Plan and the nature of the specific Health activities implemented in 2008-2011. In
doing so emerging models for both national and local adaptation and mitigation policies to address
health impacts are presented.
Plenary Session 2
Indra Vythilingam
Environmental Health Institute, 11 Biopolis Way, #06-05/08, Helios Block, Singapore 138667
Vector borne diseases are governed by many factors such as seasonal weather variation,
socioeconomic status, vector control programmes, environmental changes, deforestation and drug
resistance. In recent years much information has been churned out on climate change and its impact
of vector-borne disease epidemiology. However, the role of the vectors has always been neglected.
An important factor to consider is the increase in global travel particularly to and from vector borne
disease endemic areas. Thus, with climate change, this will increase the incidence of vector borne
diseases in temperate countries due to the non immune population and the introduction of pathogens
and vectors. Climate change will play a crucial role in the survival and transmission rate of vectors
and their pathogens. The role of these factors in relation to climate change will be discussed.
Plenary Session 3
The impact of climate change on respiratory health is due to several factors including those of extreme
temperature events, increased concentration of ground level ozone, long range air pollution resulting
from fire or aerosols, and altered distribution of allergens. Another important impact relates to the
alteration of the frequency of some respiratory infections such as respiratory syncytial virus resulting
from earlier ending of winter, or tuberculosis due to migrationwith overcrowding. Vectors relevant to
respiratory infections like water birds for avian influenza viruses will also likely to be effected by
climate change. While many will suffer some respiratory effects of climate change, those primarily
affected are individuals with pre-existing lung conditions like asthma, rhinosinusitis, chronic obstructive
pulmonary disease and lung fibrosis. The degree of impact however remains uncertain and
unpredictable, reiterating the need for better prediction models for estimation of respiratory health
impact by climate change. Low-income countries are likely to be worse affected than others. Global
efforts to mitigate climate change and to adopt adaptation measures are clearly paramount to reduce
the harm on respiratory health.
Plenary Session 4
Mak JW
International Medical University, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000 Kuala Lumpur
Concerns have been expressed globally on the deleterious effects of climate change on the environment
and human health. These effects have been documented in countries ranging from the polar region to
the tropics.
The Intergovernmental Panel on Climate Change (IPCC) defines climate change as temporal change
in climate whether due to natural variability or to human activity (IPCC, 2007). Global warming has
occurred as is evident through the increase in the average air and ocean temperatures, as well as the
melting of polar ice and snow leading to an increase in the average sea levels. The global surface
temperature (GST) 100-year linear trend (1906-2005) is 0.74 (0.56 to 0.92)C and this has increased
the ocean depths by at least 3000m, increasing the global average sea level by 3.1 (2.4 to 3.8) mm
per year from 1993 to 2003. These changes have a considerable impact on the environment.
Effects of climate change on human health can be directly due to extreme physical effects or indirectly
through their modifying effects on various determinants of the transmission cycles of infectious diseases.
Furthermore, climate change effects on human and animal behaviour need to be considered in relation
to the transmission of such diseases. However, while effects on transmission of infectious diseases
have been mostly studied, those influencing non-infectious diseases have not been addressed adequately.
Climate change effects on cancers, nutritional disorders, cardiovascular and other non-communicable
diseases will need to be addressed, but will not be covered in this review.
Emerging zoonoses in the last few decades have been attributed directly or indirectly to climate change.
It is important to know that more than 60% of microbes affecting humans have a zoonotic origin and
that at least a third of these can be further spread from human to humans after successful human
infections.
In Malaysia as in other countries, common zoonotic microbial infections include parasitic (mainly
protozoa, like cryptosporidiosis, giardiasis, microsporidiosis, etc), bacterial (leptospirosis, salmonellosis,
Escherichia coli 0157, melioidosis, etc.), and viral (Hantaviruses, Nipah virus, SARS, etc.). Most of
these are water or food-borne infections and are associated with floods, environmental degradation,
and human encroachment to wildlife domains.
This review will cover only some of the important non-vector borne zoonotic infections to avoid overlap
with the other presentations. It will discuss the direct and indirect effects of climate change on the
transmission of these zoonotic infections and the required inter-sectorial cooperation as well as the
global response needed to combat them.
Plenary Session 5
Research Molecular Biologist, USDA/ARS, 10300 Baltimore Ave BARC-East Bldg. 1042 Beltsville,
MD 20705
In previous studies we have identified The Macrophage Inhibitory Factor (MIF) in apicomplexan
parasites, E. tenella and E. acervulina that are responsible for causing coccidiosis in chickens.
Additionally, MIF from the host (Gallus gallus) was also identified. Following molecular
characterization of this molecule each of the three proteins were expressed and polyclonal antisera
were generated. This study describes the localization of MIF in the parasite as well as the host. In
the parasite, MIF appears to be present in high amounts in the developing oocyst. In the host, MIF
appears to be present in many normal tissues, suggesting that chicken MIF has functions outside of
immuno-modulation. Functional analysis of Eimeria MIF shows that the purified molecules are capable
of inhibiting the migration of chicken macrophages in vitro. Taken altogether, it is possible that at
least one of the functions of MIF in protozoa is to modulate the immune response of the host.
Plenary Session 6
Dengue fever has been known since the 18th century, but the more serious form, dengue haemorrhagic
fever, was only first known in 1952 in Manila. Today more than 100 countries are affected by dengue
with 50-100 million cases yearly. There are about 500,000 hospitalisation due to dengue yearly and
about 2.5% of cases die. In the continued absence of an effective tetravalent vaccine and specific
treatment, dengue can only be controlled via suppression and elimination of the Aedes vectors. Present
vector control technology, however, appears to be ineffective to control the disease, often resulting in
massive outbreaks. There is a need to search for more effective dengue vector control technology
such as the possible use of genetic control in which sterile males are released to mate with the wild
type females, resulting in death in the larval stage, thereby sustained relaese will be able to suppress
the natural population to level below the transmission threshold. Recent advances in biotechnology
have resulted in a strain of Aedes aegypti habouring a lethal gene killing the larvae. The technique,
known as Release of Insects with Dominant Lethality (RIDL) has been investigated in the Institute
for Medical Research (IMR) since 2006 under the IMR-Oxitec Joint Initiative. Extensive laboratory
studies indicated the bionomics, life history, vectorial capacity (dengue and chikungunya), insecticide
susceptibility, oviposition behaviour, interspecific mating behaviour and horizontal gene transfer were
all similar to the wild type; showing the introduced RIDL gene has not affected other biological functions
of the mosquito. In semi-field trial, the mating competitiveness of RIDL Ae aegypti males was similar
to that of wild type. These findings indicated that genetic control of Ae aegypti was promising and
field release application was subsequently sought. The first release to study the longevity and dispersal
of the RIDL Ae aegypti was successfully conducted on 21 December 2010 in a forested uninhabited
area after following an extensive process described by my collaborator. Preliminary analysis of results
reveals that the two strains have comparable daily survival probability, while the OX513A strain does
not disperse quite as well as the unmodified laboratory strain even though the farthest point of recapture
was similar for both strains.
Seshadri Vasan
Dengue is the fastest-growing vector-borne disease of the world. In Malaysia, for instance, reported
cases have increased dramatically from 7103 cases in 2000 to 46171 cases in 2010. Several countries
are evaluating the biosafety and efficacy of the RIDL-Sterile Insect Technique a new tool developed
at Oxford University (United Kingdom) and its part-owned company Oxitec to suppress the Aedes
aegypti vector population and thus combat dengue and chikungunya. As of 2010, Brazil, Cayman Islands
(UK Overseas Territory) and Malaysia have approved open field demonstration, with open releases
having taken place in the latter two countries. Since 2009, Cayman Islands authorities have released
around 3.3 million male mosquitoes of this strain in inhabited locations and reported 80% suppression
in vector population. Malaysia conducted a limited release the first open release outside the UK
and its Overseas Territories in an uninhabited forest in December 2010, four years after it became
the first endemic country in the world to import the OX513A strain for contained evaluation. In the
intervening years, the Institute for Medical Research (IMR) has conducted extensive evaluation of
this strains biosafety and efficacy, including the worlds first semi-field trials (2007-08), dedicated
workshops on risk assessment (2008) and risk communication (2010), and extensive national consultation
with numerous committees, the public and NGOs (2009-10). IMRs MRR experiment in the uninhabited
site seems to indicate that the genetic modification has not adversely affected survival of adult male
mosquitoes in the field; however, it is necessary to repeat this experiment in inhabited locations (the
natural habitat of Aedes aegypti) to reconfirm this encouraging result. These steps laboratory and
semi-field trials, MRR trials in uninhabited and inhabited sites, followed by suppression trials represent
a measured series of experiments of increasing scale and sophistication; this is widely viewed as the
appropriate way to evaluate a new technology for potential field use, and is consistent with a step-
wise approach advocated by regulatory authorities worldwide prior to deployment of an area-wide
control programme.
Oral Session 1
OS1.1
Stable fly Stomoxys calcitrans is one of the blood-sucking flies prevalent in tropical and subtropical
countries which is resemble to house fly Musca domestica (Family Muscidae). Two generations of
stable flies were used at age 24 and 48 hours. The flies were fed on infected mouse blood with
Trypanosoma evansi through chickens skin placed on glass container in water bath at 37C. The
fed flies were killed by chloroform and dissected under the stereo-microscope at different intervals:
immediate, 1, 12, 24, 26, 27, 28, 29 and 30 hours post-feeding (ten flies for each interval). Smears
were prepared from isolated trypanosomes and then inoculated via intraperitoneally to four mice. Three
serological tests were employed to detect the antigenic variation of T. evansi namely agglutination,
indirect immunoflurescent antibody (IFAT) and gel diffusion. All mice were positive for T. evansi for
all intervals except 27, 28 and 29 hours post-feeding. The parasites were disintegrated at 30 hours of
exposure. Sign of longitudinal binary fission was observed at 12 hours post-feeding. Indirect
immunofluorescent antibody test showed cross-reaction between different isolates of T. evansi in
comparison with agglutination and gel diffusion tests. In conclusion, the infectivity was decreased with
time of feeding and this might be correlated with the loss of external coat of the parasites. In addition,
the depletion of nutritional supply in the blood might leads to disintegration of the parasites. In IFAT,
the cross reactivity was attributed to the specific and common antibody-antigen reactions.
OS1.2
Bacillus thuringiensis israelensis (Bti) is a naturally occurring soil bacterium registered for mosquito
larvae control which is commonly used as larvicidal agent. Trypsin modulating oostatic factor (TMOF),
a hormone that stops mosquitoes from producing enzyme called trypsin, preventing them to draw
nutrients from food causing them to starve to death. This study was conducted to evaluate the
effectiveness and residual effect of 4% TMOF + 4% Bti rice husk, 2% TMOF + 2% Bti rice husk
and TMOF-Bti wettable powder formulations on Ae. aegypti larvae at UKM Campus Kuala Lumpur.
20 first instar Ae. aegypti larvae were placed in a plastic bucket containing 4 liters of water supplied
with crushed dried leaf powder as their source of food. Combination of TMOF-Bti in rice husk
formulation with the following weights viz: 10 mg, 25 mg, 50 mg and 100 mg in duplicate were
distributed in the buckets; while TMOF-Bti in wettable powder formulation each weighing viz; 2mg, 5
mg, 10 mg and 20 mg in duplicate were also placed in the buckets. The control buckets runs in duplicate
with 4 liter of water and 20 first instar Ae. aegypti larvae. All buckets were covered by mosquito
netting. Larval survival was recorded after 24 hours and weekly for five weeks. A new batch of 20
1st instar larvae Ae. aegypti was introduced into the buckets weekly without additional TMOF-Bti
rice husk formulation or wettable powder. The result of the study showed that all formulations were
very effective on the first two weeks by giving 0% larval survival for all concentrations applied. On
the third week to fifth weeks, the larval survival increased gradually from as high as 70% for the 24
hour larval survival on the fifth week and dropped to 30% after a week in the fifth week. Thus, all
these 3 formulations of TMOF-Bti could retain their residual effect on killing the first instar Ae. aegypti
larvae for 5 weeks at least effectively.
OS1.3
Oviposition site selection behavior of the gravid female of a laboratory strain of Aedes aegypti (L)
was studied to understand the natural pattern of breeding preference in the field. Three sets of
experiment were conducted: First set was to determine the number of eggs laid by a single gravid
female mosquito released into a cage containing only one ovitrap; second experiment was conducted
to examine the oviposition behaviour when the mosquito was given a choice to oviposit. In the later,
four oviposition cups i.e.: without larvae, with low, medium and high number of larvae at L1, L2, L3
and L4 instar, respectively. Similar experiment was also conducted using the pupae A gravid Aedes
aegypti was introduced into the cage and the oviposition was determined after 24 hours. The third
experiment was conducted to determine the oviposition behaviour in the presence and absence of
paddle in the ovitrap. The results of the first experiment indicated that a female gravid Aedes aegypti
deposited all her eggs into the ovitrap on the first day of oviposition. In the second experiment, skip
oviposition was observed when different oviposition choices were provided. Generally it preferred to
oviposit in container without L1 larvae, without L2 larvae or with low number of L2 larvae, high number
of L3 larvae, low number of L4 larvae and without pupae or with low number of pupae.. Finally,
there was no significant difference in the oviposition behaviour in the absence or presence of paddle
which did not play an important role in ovitrapping.
OS1.4
Kim-Sung Lee1, Sharon Lo1, Yee-Ling Lai1, Tim Barkham2, Pauline Aw3, Peng-Lim Ooi4,
Ji-Choong Tai5, Martin Hibberd3, Edward C Holmes6 and Lee-Ching Ng1
1National Environment Agency, Singapore
2Tan Tock Seng Hospital, Singapore
3Genome Institute of Singapore, Singapore
4Ministry of Health, Singapore
6Pennsylvania State University, USA
Singapore is hyperendemic for dengue, with all 4 serotypes found circulating in the island. Dengue
serotype 2 (DENV-2) has been the predominant serotype since 2007 after it replaced DENV-1 and
caused a major outbreak in that year. Analysis of the envelope (E) protein gene showed that the 2007
outbreak was also accompanied by a clade replacement within the Cosmopolitan genotype; new clade
viruses replacing the old clade viruses (sampled before 2007). Interestingly, sequence analysis of recent
samples showed an increased in viral diversity within the new clade DENV-2, with two distinct lineages
emerged as a result of local viral evolution. Phylogeographic analysis of the complete genome of new
clade DENV-2 also revealed varying degrees of viral diversity at spatial level. Viruses originating
from the central region had higher level of genetic diversity, suggesting that the transmission intensity
was relatively higher compared to other parts of the island. The trend of mixing and exchanges of
DENV-2 indicates that the central part was highly likely the mixing ground for hyperendemic
transmission. In addition, our virological surveillance revealed multiple importations and co-circulation
of various strains of DENV-2, DENV-3 and DENV-4 that were uncommon to Singapore. In particular,
phylogenetic analysis of DENV-3 showed substantial diversity with multiple strains that were closely
related to dengue viruses previously reported around the Asian region. These findings highlight the
importance of viral migration and evolution in dengue epidemics and suggest that cross-border
surveillance programme and regional collaborations are needed in an effort to control the disease.
OS1.5
Nurhainis Ogu Salim, Karnan G, Siti Nor Fatimah AH, Synthia Molupin, Adio Babatunde Wasiu
and Shamilah Hisam
Parasitology Unit, Infectious Disease Research Center, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur
Animals do harbor a significant number of parasites that are transmissible to man either by direct
ingestion of eggs (fecal-oral route) or skin penetration by larval forms of the parasites. Personal hygiene
certainly plays a major role in ensuring accidental infections do not take place. In an environment
where animals and human mingle, such as in close relationship between pets and owners, unintentional
infections could still take place. We assume the risk of acquiring infection is minimal since owners
take their pets for regular de-worming and vaccination. This study was aimed to determine the
prevalence of parasitic infections among dogs and cats from two different environments: domestic
(pets) versus stray animals with the assumption that regular de-worming and/or vaccination minimizes
the parasitic infections among dogs and cats of domestic origins as compared to stray animals. Stool
specimens of dogs and cats from Dewan Bandaraya Kuala Lumpur (DBKLs) animal pound in Jalan
Setapak and SPCA Jalan Ampang temporary shelters for animals and from 3 veterinary clinics in
Shah Alam, Taman Tun Dr Ismail Jaya and Jalan Gasing were collected and examined by stool
concentration methods and microscopy. Stool examination showed a higher prevalence rate of intestinal
parasitic infections among stray dogs and cats as compared to domestic cats and dogs, with 58.6%
versus 28.1%, p < 0.001, respectively. The most common parasitic infections found in these dogs and
cats were hookworms (25.1%), Ascaris lumbricoides (21.1%), Trichiuris trichiura (18.3%) and
Giardia lamblia (0.6%). No other intestinal protozoas were detected. In conclusion, results showed
a higher percentage of ova and cysts in stools of dogs and cats of homeless origin compared to
domesticated ones. Our postulate is that vigilant veterinary management and owner awareness of
animal health had played a significant role in minimizing parasitic infections in pets and owners should
continue their animal regular medical check-ups and antihelminthic/vaccination programs.
OS1.6
Adela IJ1, Shamilah Hisam1, Ooi CH2, Nor Parina I1, Siti Zulaikha Hussin1,
Wan Noor Ayuni WN1, Synthia Molupin1, Yusri MY1 and Lokman HS3
1Parasitology Unit, Infectious Diseases Research Centre, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur, Malaysia
2Vector Control for Vector-Borne Diseases, Sarawak Health Department, Jalan Tun Abang Haji Openg,
Malaysia
Malaria incidence in Malaysia is still under control with 4,484 cases in 2010 (from January to August,
VBDCP, 2010). The highest endemic state affected by this disease is Sabah and the most prevalent
malaria species is Plasmodium vivax (56%). Since the report of naturally acquired P. knowlesi in
Kapit, Sarawak (2004), the Malaria Control Programme in Sarawak has diligently screened all blood
films for P. knowlesi from all divisions. As a result, in 2009, Sarawak had a large number of BFMPs
identified microscopically as P. knowlesi / P. malariae. However, due to morphological similarities
between P. knowlesi, P. malariae and P. falciparum, a total of 809 BFMPs earlier microscopically
confirmed as Pm/Pk by Sarawak SH were sent to IMR (Malaria Reference Centre for the Ministry
of Health) for re-confirmation. All BFMPs were re-examined by trained microscopists in IMR for
identification of malaria parasites, estimation of parasitaemia and molecular confirmation. A nested
PCR as described by Singh et al was used to identify P. knowlesi and human malaria parasites
infections. 377 slides completed re-examination by PCR and microscopy. By microscopic re-
examination, 326 blood films were identified as Pm/Pk by IMR, followed by P. malariae (19/377)
and other malaria species (32/377). By PCR, 30.5% out of 377 blood slides were positive and 69.5%
were PCR neg. An approximate 59% of blood films identified as Pm/Pk by microscopy were found
to be P. knowlesi by PCR and surprisingly, 60% of blood films earlier identified as P. malariae turned
out to be P. falciparum by PCR and only 20% were Pk by PCR and none of blood films were Pm by
PCR. This results shows that identification of P. knowlesi by microscopy alone could be misleading.
Although PCR is still needed, it is highly dependent on the quality of DNA received. In contrast,
identification by microscopy is highly dependent on the skills of microscopists and the quality of smears
being made.
OS1.7
Nurulhusna AH1, Khadri MS1, Sisma ST3, Abdullah AG1, Hisyamuddin AH2, Norazlina AH1,
Khairul-Asuad M1, Muhammad-Azim, AK1 and Lee HL
1Medical Entomology Unit, Infectious Diseases Research Centre, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur, Malaysia
2VectorBorne Disease Control Unit, Pejabat Kesihatan Daerah Melaka Tengah, Jalan Bukit Baru,
75150, Malacca
3School of Diploma in Applied Parasitology and Entomology, Institute for Medical Research, Jalan Pahang,
Study on effectiveness of the IMR Autocidal Trap in controlling field Aedes mosquito population was
conducted from 8th July to 3rd Sept 2010 in Malacca Tengah viz. Taman Peringgit Jaya and Taman
Kenanga. The autocidal trap device is a modification of traditional ovitrap and is capable of monitoring
and controlling Aedes species. The trap is equipped with a floater fitted with copper wire-mesh and a
transparent round sticky strip that can trap not only the larvae but also the adults. Resident in 85
houses out of a total of 1761, (5%) houses in both selected areas participated in this study, after informed
consent was obtained. Each house was installed with three autocidal traps. The traps were filled with
tap water and placed at suitable location indoor and outdoor of the house. Every two weeks the sticky
strip was collected and the insects trapped on the strip were identified and recorded. The results showed
that three main species of mosquitoes were trapped on the sticky strip: Aedes aegypti (L.), Aedes
albopictus Skuse and Culex quinquefasciatus Say. Female Ae. aegypti was the highest mosquito
group trapped (43%) on the sticky trap, followed by female Cx. quinquefasciatus (18%) and female
Ae. albopictus (6%). Because of the rainy season in Malacca during the study, 20% of the mosquito
species were damaged and could not be identified. A total of 18 houses (21%) were positive with
Aedes mosquitoes. Therefore, a mean of 4 Aedes adults were trapped/sticky strip/2 months/house
with the maximum 22 Aedes sp. were caught in one sticky strip. Observation under microscope showed
Aedes eggs were deposited onto the sticky strip, indicating that gravid female mosquitoes were attracted
to lay their eggs in the autocidal trap. Twenty two percent out of 827 mosquitoes trapped in the
autocidal trap was Culex species. The IMR Autocidal Trap is therefore an effective mosquito trapping
device.
OS1.8
Oreenaiza MN1, Wesly D 1, Wong HM 1, Teoh GN 1, Khairul A1, Nor Azlina AH1,
Azahari AH 1, Muhammad ZS1, Renaud L2, Derric N3, Nazni WA1 and Lee HL1
1Medical Entolology Unit, Institute for Medical Research, Jalan Pahang 50588, Kuala Lumpur
2Malaysia Oxitec S/B, Plaza See Hoy Chan, Suite 1502, Jalan Raja Chulan, Kuala Lumpur, 50200, Malaysia
3Oxitec Limited, 71 Milton Park, Oxford OX14 4RX, United Kingdom
Dengue continues to cause high human morbidity and mortality compared with any other vector-borne
viral diseases. To date, there has been no specific treatment and vaccine. Thus, it is evidently essential
to develop new and effective methods, such as genetic control. Recently, the development of RIDL
(Release of Insect carrying Dominant Lethality) mosquitoes, particularly the release of males carrying
this dominant lethal gene as part of this integrated tool in vector control is being evaluated. However,
prior to deployment of these sterile males, it is essential to study the biology and behavior of the closest
related predator species in order to determine the possible effect of gene transfer on life span, size,
longevity and fecundity of the predator, Tx. splendens (Thailand) and Tx. amboinensis (Hawaiian)
on Wild Type (WT), and transgenic Ae. aegypti larvae bred on tetracycline and without tetracycline.
The results indicated that the life cycle, wing-length and fecundity of Toxorhynchites species fed
with transgenic and WT Ae. aegypti larvae remained unchanged. Further test of Toxorhynchites sp.
larval stage 4 of F1 generation using polymerase chain reaction (PCR) showed negative results,
indicating absence of lethal gene being transferred via predation. Thus, horizontal gene transfer of
the lethal gene to Toxorhynchites larvae via ingestion is extremely unlikely.
Oral Session 2
OS2.1
Division of Acarology, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
Scrub typhus has been recognized in numerous countries in Asian-Pacific area including Malaysia;
the disease was first noted in Malaysia in 1915 by Dowden. This project is to detect the presence of
scrub typhus in preserved and live chiggers and screened host target organs such as liver, spleen as
well as blood. All samples were processed by a standard method that began with extraction of DNA,
then continued with polymerase chain reaction (PCR) and followed by gel electrophoresis. A total of
465 chiggers (Leptotrmbidium deliense) and 7 different species of hosts (Ratus diardii, Tupaia glis,
T. minor and R. tiomanicus, Sundamys muelleri and R. whiteheadi, Leopoldamys sabanus) were
collected from 12 different locations (Bukit Sagu; Bukit Goh; Bukit Kuantan; Chengal Lempung, Balok
and Tg. Lumpur-Kuantan, Pahang, Bukit Panchor and Pulau Jerjak-Penang, Ketengah Jaya and Setiu;
Terengganu, Pulau Pangkor, Perak and Sungai Sedim, Kedah) and tested. Collection sites included
oil palm plantation, rubber estate, secondary forest, housing estate and coastal area. All specimens
had tested negative except for a blood sample from L. sabanus from Sungai Sedim, Kedah.
OS2.2
Nor Azrina Norahmad1, Noor Rain Abdullah, Norhayati Yaccob, Jenarun Jelip2, Jiloris F Dony3,
Lokman Hakim Sulaiman3, Hasidah Mohd Sidek4, Harald Noedl5 and Zakiah Ismail1
1Herbal Medicine Research Center, Institute for Medical Research, 50588, Kuala Lumpur
2Sabah State Health Department, Federal House, Jalan Mat Salleh, 88590, Kota Kinabalu, Sabah
3Disease Control Division, Ministry of Health Malaysia, Block E10, Complex E, 62590, Putrajaya
4School of Biosciences and Biotechnology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia
5Medical University of Vienna, Kinderspitalgasse 15, A-1090 Vienna, Austria
Sulfadoxine and pyrimethamine (SP) is currently the first line drug for the treatment of falciparum
malaria in Sabah, Malaysia. Resistant of P. falciparum to SP has been observed in Malaysia since
1960s. The quintuple mutations have previously been recognized to strongly predict clinical outcome
of SP resistance. The objective of this study was to determine the present of point mutation in pfdhps
and pfdhfr genes from 31 microscopically confirmed blood samples of individuals from malaria endemic
areas in Kalabakan, Tawau. Nested mutation specific polymerase chain reaction and restriction digestion
methods were used to detect the presence of mutations at 16, 51, 59, 108 and 164 codon position in
pfdhfr gene and mutations in the 437, 540 and 581 codon sites in pfdhps gene. We found that all
samples had point mutations in at least one codon of pfdhfr gene. Changes restricted to codons 16V,
108N and 164L were 16.1%, 67.7% and 80.6% respectively while mutation at codon 59R was found
in all isolates. No mutations were detected at 51N and there were no changes from 108S to 108T.
With regards to pfdhps, we found that all of the isolates had the 437G mutation. High proportions
(74.2%) of the samples have mutation at 581G codon position. However all the samples showed wild-
type for codon 540. Although there were no quintuple mutations found in the isolates, the high number
of samples with triple mutation (59R108N/437G) may indicate that the parasite within this population
could have the potential to develop into quintuple mutants in the future.
OS2.3
Atique Ahmed1, Paul CS Divis1, Lu Chan Woon3, Balbir Singh1, Sanjeev Krishna1, 2 and
Janet Cox-Singh1,2
1Malaria Research Centre, University Malaysia Sarawak, Kuching, 93150, Malaysia
2Centre for Infection and Immunity, St Georges University of London, London SW17 0RE, UK
3Pathology Laboratory, Hospital Sarikei, Sarikei, 96100, Malaysia
Plasmodium species cause malaria by invading and multiplying inside the host red blood cells. The
parasite uses multiple invasion ligands which bind to specific red cell receptors. The relative efficiency
by which these ligands interact with RBC receptors may be critical to the rate of increase in
parasitaemia. Hyperparasitaemia is associated with severe disease in Plasmodium knowlesi, a zoonotic
malaria, found in Southeast Asia. The aims of the study are to test the hypothesis that Plasmodium
knowlesi invasion gene haplotypes have an association with parasitaemia at presentation and clinical
outcome. A gene encoding the Plasmodium knowlesi normocyte binding protein xa (Pknbpxa) was
chosen for this work because it is orthologous to invasion gene families encoding reticulocyte homologs
of P. falciparum and P. vivax. An amplification protocol for the Pknbpxa gene was developed and a
method for cloning and sequencing was optimized. The full length sequence of Pknbpxa (8,540bp)
has been obtained from one field isolate of P. knowlesi. A reliable haplotyping assay will be developed
following sequencing of other field isolates to screen for polymorphisms in the Pknbpxa gene from
knowlesi malaria patients with known parasitaemia and disease severity.
OS2.4
Rahmah N1, Jenarun J2, Mohd Hafiz AH3, Lokman H3, Won K4, Chu B5,
Muhammad Hafiznur Y1 and Zulkarnain MI1
1Institutefor Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia
2Sabah State Health Department, Sabah, Malaysia
3Ministry of Health Malaysia, Putrajaya, Malaysia
4Centres for Disease Control, Atlanta, USA
5Task Force for Global Health, Atlanta, USA
An operational research was performed in Sabah, Malaysia with the objective of determining whether
the annual mass drug administration (MDA) of the National Programme for the Elimination of
Lymphatic Filariasis can be stopped or should be continued. The main diagnostic tool used was a lateral
flow cassette test called panLF rapid, an IgG4-based assay using two filarial recombinant antigens,
BmR1 and BmSXP. Day-time finger prick blood samples from 2437 individuals were sampled and tested
in the field by panLF rapid, 1430 were from 7-8 year old school children and 1007 were from adults.
A total of 90 children (6.3%) and 132 adults (13.1%) were found to be positive by the rapid test.
Positive individuals were further sampled at night to test for presence of microfilaria (mf) by thick
blood smear and real-time PCR. A total of 31/87 (35.6%) children and 29/128 (22.7%) adults were
found to be mf positive. Real-time PCR was positive in 52.9% (46/87) children and 28.9% (37/128)
adults. The study showed that the prevalence of brugian lymphatic filariasis in the area was still high,
notably in children. Subsequently the decision was made to continue the MDA for at least two more
cycles.
Oral Session 3
OS3.1
Assessments of potential effects of climate change and other environmental dynamics on spread of
zoonotic pathogens must rely on long-term biomonitoring programs that accurately sample environments
for pathogens across periods adequate to detect correlation with changing environmental conditions.
From 1990-2020, we developed long-term monitoring programs for the opportunistic protists,
Cryptosporidium parvum and Giardia lamblia, and the emerging human-virulent microsporidia,
Enterocytozoon bieneusi and Encephalitozoon spp., in major freshwater systems and agricultural
regions of North America and Europe. Because these and other waterborne pathogens are prevalent
throughout the world, we propose expanding such monitoring efforts to other areas, including Malaysia
and elsewhere in Southeast Asia and the Indo-Pacific region, with modifications to identify local sentinel
organisms. Our methods couple use of sentinel organisms for field collection with laboratory diagnosis
using molecular markers. Across a range of climate conditions, we accurately detected each of these
pathogens in up to 93% of samples in major rivers and smaller watersheds in both urban and
agriculturally intensive areas. Sentinel organisms found to be effective include filter-feeding molluscs,
coprophagous flies, and dung beetles. We found all to accumulate and retain viable pathogens for
extended periods, making them available for diagnosis using staining, PCR, and/or Fluorescent In-Situ
Hybridization (FISH) with immunofluorescent monoclonal antibody tags (IFA).
OS3.2
Saudi Arabia
Schistosomiasis the most important human helminthes infection is a major source of morbidity for people
in 76 countries. Schistosoma mansoni and Schistosoma haematobium are endemic in Saudi Arabia.
The epidemiology of the disease, the snail intermediate hosts and the governmental successful control
efforts are covered in the form of a review. In addition, the major challenges for control of
schistosomiasis in the country are discussed.
OS3.3
Cryptosporidium sp. was detected in 3 cows from rectal pinch samples. Direct smear stained with
Acid Fast and Kinyoun stain was used to detect the organism. Subsequent samplings also indicated
positive for Cryptosporidiosis, whereby one of the animals died due to dehydration and severe clinical
signs of diarrhoea. The farm had contaminated water supply where 2 out of the four ponds were
positive for Cryptosporidium sp. whereas the municipal water supply was negative. The management
of the farm was poor in terms of nutrition and cleanliness which led to Cryptosporidium sp. infection
in the cattle compounded by stress factors. The mortality of the adult dairy cattle and calves was
also high reaching up to 40%. The most common cause of death was leg weakness, severe dehydration
and pneumonia in calves as a result of severe infections. Cryptosporidiosis is zoonotic and thus needs
to be controlled to prevent outbreaks in the human population.
Oral Session 4
OS4.1
Kavitha Rajagopal, Tan Tian Chye2, Lee Han Lim3 and Mohd Sofian Azirun
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia
2Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur
3Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
Forensic entomology applies knowledge about the behaviour and ecology of insects associated to
corpses to crime scene investigations. It is possible to calculate a minimum post-mortem interval (PMI)
by determining the age of the oldest blowfly larvae feeding on a corpse. It is, thus, crucial to correctly
identify the species collected from crime scene investigations. This represents the first study in Malaysia
to identify blowfly species based on specimens collected during crime scene investigation. We evaluated
both morphological and molecular tolls in species identification for 11 individuals of 4 different species
sampled from 10 different crime scenes in Malaysia. A molecular identification method involving the
sequencing 2.4 kilo basepairs barcode fragment of the COI, COII and t-RNA leucine genes from
11 specimens, representating 10 different crime scenes. A phylogenetic tree is generated using a
neighbour-joining technique. This study confirmed the presence of Chrysomya megacephala,
Chrysomya rufifacies, Chrysomya nigripes and Lucilia cuprina. Chrysomya megacephala was
the predominant species found. In addition, we identified a new species based upon complete genes
sequencing.
OS4.2
Chew WK1, Nazni WA1, Farah Hasbolah1, Heo CC2, Heah SK1, Hiromu Kurahashi3 and
Lee HL 1
1Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
2Faculty of Medicine, Universiti Teknologi Mara (UiTM), 40450 Shah Alam, Selangor, Malaysia
3International Department of Dipterology (IDD), Hikawadai 1-2-2-1, Higashikurume City, Tokyo 203-0004
Japan
Forensic entomology is the study of insects and other forensic arthropods associated with legal
investigation. Often, the location of a cadaver may affect the diversity of forensic arthropods. The
objective of this study was to determine the diversity of arthropods fauna in a high rise building. The
study site was a 26th storey building in Sentul Timur, Kuala Lumpur. Fresh cow liver and meat, used
as bait to attract the flies, were placed in an indoor and outdoor location on the 26th storey of the
study building. The duration of the study was 21 days until no more flies were observed. The arthropods
infesting the baits were collected and identified as Megaselia scalaris (Loew), Chrysomya
megacephala (Fabricius), Boettcherisca peregrina (Robineau-Desvoidy), Parasarcophaga dux
(Thomson), Musca domestica Linnaeus, ants, and spider. In addition, fly maggots were collected and
mounted for identification. The maggots were those of Megaselia scalaris, and species of the family
Sarcophagidae. The adults and larvae were collected both from indoor and outdoor. The study also
showed that flies were attracted to the baits within 8.82 0.42 hours in a high rise building, indicating
that cadaver infestation can occur within the same day of death in a high rise building.
OS4.3
Nazni WA1, Lee HL1, Chew WK1, Azahari AH1 and Disney RHL2
1Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
2Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ England
Forensic entomology is the application and study of insect and other arthropods related to legal matters.
Forensic entomology can be divided into three subfields: urban, stored-product and medico legal/
medico-criminal entomology. In this study we reported for the first time the involvement of insects in
an industrial sector and we termed it Industrial Forensic Entomology. In this case, maggots were found
breeding at the inline filter of the liquid cream yeast circulation pipeline of an established bread producer.
The investigation was to determine if the fly had infested the yeast at the yeast production industry
(YPI) or the infestation began at the bread producer industry (BPI), as this has legal implication.
Specimens of maggot were collected and sent to us for identification and to determine if the identified
maggot species could survive and develop at or below 6C, at which temperature the yeast was stored.
The maggot specimens were identified as 3rd instar Megaselia scalaris (Leow). We conducted a
study on the development of M. scalaris in the laboratory at room temperature and below 6C. Study
at room temperature indicated that the complete life cycle of M. scalaris from egg to adult in liquid
yeast media was 14-16 days. The eggs hatched within 12 h, while the larval and pupal period was
about 60 h and 10 days, respectively. Male flies emerged before females, while the first oviposition of
females was about 2 days after emergence. Megaselia eggs could not hatch at low temperature at
4-5C in another experiment, using first instar larvae only about 8% of larvae could survive for a
maximum of 3 days but these larvae failed to pupate and develop into adults. These data indicated
that Megaselia could not breed in yeast stored continuously at low temperature.
Oral Session 5
OS5.1
This study was conducted to determine the toxicity of two commercial disinfectants containing
chloroxylenol and benzyl chlorophenol, against dust mites, Dermatophagoides pteronyssinus and
Dermatophagoides farinae in the laboratory. The contact, topical, and residual activities as well as
simulated assay of the two commercial disinfectants were investigated at five concentrations (2.4%,
1.2%, 0.6%, 0.3% and 0.15%). For contact and residual activities, mites were exposed to Whatman
No. 1 filter paper impregnated with disinfectants solution while for topical activity, disinfectant solution
were applied direct to the dust mites. For simulated test, mites were exposed to cotton fabric
impregnated with disinfectant solution. Mortalities from chloroxylenol solution were better than benzyl
chlorophenol for all activities. At 2.4% concentration, 24 hrs contact exposure to chloroxylenol resulted
in mortality greater than 90% while mortality was more than 50% for topical exposure in both species
of dust mites. 100% mortality was recorded for 24 hrs post treatment of residual exposure to 2.4%
chloroxylenol in both species of dust mites but the mortalities of D. pteronyssinus declined to 70%
by week 4. Meanwhile, 2.4% benzyl chlorophenol at 24 hrs contact and topical exposures produced
higher mortalities in D. pteronyssinus than D. farinae; however the mortalities in D. farinae were
greater than D. pteronyssinus for 24hrs residual exposure at that concentration and mortalities for
both species declined progressively by week 4. Mortalities of D. pteronyssinus for simulated test of
chloroxylenol were better than D. farinae while mortalities for both species were similar for benzyl
chlorophenol.
OS5.2
One of the major constraints to the small ruminant industry in Malaysia is parasitic gastroenteritis.
The main form of control for many years lies in anthelmintics hence the nematode parasites have
developed anthelmintic resistance, reported to be widespread in Malaysia. This study was done to
evaluate the present anthelmintic resistance status of goats in Terengganu. A total of 141 goats from
six farms were chosen for detection of resistance using the Fecal Egg Count Reduction Test, using
levamisole, ivermectin, benzimidazole and closantel. Fecal samples taken were subjected to fecal egg
count and larval culture for third stage strongyle larvae identification. All farms had resistance towards
benzimidazole and closantel while only two farms were still susceptible to levamisole and one farm
had suspected resistance to ivermectin. There were four out of six farms that had resistance to all
anthelmintics tested. The strongyles which had developed anthelmintic resistance were predominantly
Haemonchus sp. followed by Trichostrongylus sp. The findings of this study showed that anthelmintic
resistance in Terengganu has escalated and the need for effective immediate action in particular with
regard to farmer education is very important to salvage the small ruminant industry.
OS5.3
Lagrosa Gino Antonio C1, Kurosaki Mayumi J1, Labajosa Paul Vincent B1,
Lacap Maria Rizza N1, Langurayan Geneva Faye E1, Lantin Kharen U1,
Matienzo Evangeline T1, Porto Analin E1, Paat JN Junnile S1 and Albano Pia Marie SP1,2
1College of Nursing, University of Santo Tomas, Espaa, Manila, Philippines
2Research Center for the Natural and Applied Sciences, Thomas Aquinas Research Complex,
University of Santo Tomas, Espaa, Manila, Philippines
Parasitism remains a public health problem in the Philippines despite deworming programs by the
government. This study aimed at identifying the factors that contribute to this issue. Routine fecal
examination was done on members of 46 families from 2 different depressed areas in Metro Manila,
Philippines. Results were then correlated with the health practices, environmental, and sanitation
conditions of the families involved in the study. Residents from the dumpsite area were found to have
higher prevalence of intestinal parasitism (76%) compared to coastal area dwellers (41%). While
Ascaris lumbricoides and Enterobius vermicularis infections were prevalent in the dumpsite area,
fecal samples of residents from the coastal area were positive mostly for monoecious flukes and even
cestodes. Unsatisfactory storage of drinking water and irregular house cleaning were demonstrated
as major factors for the persistence of intestinal parasitism regardless of deworming the dumpsite
residents. Conversely, the coastal area dwellers lacked latrine in their homes and practiced improper
garbage segregation and excreta disposal. They also admitted to serving sun-dried seafoods, which
they would catch from the water just below or near their houses. Moreover, their unconcern about
their last deworming time proved to contribute to the prevalence of parasitism in their community.
Hence, constant deworming was not sufficient to reduce incidence of parasitism in such communities.
It is, therefore, most important to break the transmission cycle of the parasite through proper education
and monitoring of the health practices, environmental and sanitation conditions especially of the urban
slum dwellers.
OS5.4
Prolonged state of oxidative stress (caused by overproduction of free radicals than antioxidants) has
been one of the contributory factors of cancer. Free radicals that are generated by hosts immune
cells to kill the invading parasites implicate parasitic infections as a possible cause of oxidative stress.
The activity of free radicals namely reactive oxygen species (ROS) can cause secretion of metabolites
in serum such as malondialdehyde (MDA, metabolite of lipid peroxidation), hydrogen peroxide (H2O2,
indicates hydroxyl radical level) and AOPP (advanced oxidation protein product). Previously, we have
reported that colorectal cancer patients (majority with B. hominis infection) exhibited high level of
AOPP. This indicates that the possibility of intestinal parasitic infection in facilitating cancer growth
via oxidative protein damage should not be ruled out. However such studies have not been reported
in breast cancer. This study compares MDA, H2O2 and AOPP in subjects infected with intestinal
parasites alone and breast cancer patients with and without intestinal parasitic infection. All intestinal
parasite infected subjects and breast cancer patients showed high level of oxidative stress compared
to the healthy individuals. The levels of H2O2 in breast cancer patients with infection were significantly
higher compared to patients without infection. This implicates that intestinal parasitic infections in cancer
are generally detrimental regardless of the cancer types. To date this is the first study to report on
the effect of intestinal parasitic infection towards oxidative damage in breast cancer. Thus, cancer
patients who are mainly known for immunodeficiency should be subjected for intestinal parasitic
screening.
OS5.5
Mariana A1, Shukor MN2, Muhd Norhazizi H1, Intan Nurlemsha B1 and Ho TM1
1Acarology Unit, Infectious Diseases Research Centre, Institute for Medical Research, 50588 Kuala Lumpur,
Malaysia
2School of Environmental and Natural Resources Sciences, Faculty of Science and Technology,
OS5.6
Rosilah AA1, Godard M2, Walker A2, Williams C3, Aprahamian M3 and Brooks D1
1School of Environment and Life Sciences, University of Salford, Salford, UK
2CEFAS, Lowestoft, UK
3Environment Agency, UK
Like other living things, fish also suffer from diseases and parasites. Parasitic infections lead to serious
consequences of fish behavior where they cause morbidity, mortality and economic losses in fish
production in the world. The European eel, Anguilla anguilla, is an economically important species
contributing to the biodiversity of UK and European inland and coastal waters. However, the
recruitment of European eel reported to suffer a great losses and one contributory factor is believed
to be the effect of infection with parasites. The swim-bladder nematode Anguillicoloides crassus,
has gained much attention more than other eel parasites since its introduction into Europe in early
1980s. This is due to its relatively recent and rapid invasion of European waters and the subsequent
debilitating pathology that occurs within A. anguilla. However, the natural host, the Pacific eel A.
japonica, is not harmed by A. crassus infection. In this study, we seek to clarify the status of A.
crassus in UK habitats. Such information is necessary as a basis for the control of spreading of this
parasite and for future improving the management of European eel in UK. Data highlighting the
prevalence and intensity of A. crassus infection in UK eel populations will be presented at the meeting.
OS5.7
Six local female horses aged between three to seven years old were used to study the dynamics of
Trypanosoma evansi infection in horses. Four animals were injected with 102 parasites/ kg body weight
intravenously, while the remaining two animals served as negative control. Blood samples and rectal
temperature were collected on alternative days from both groups for fifty days. The infected group
was salvage treated on day 31 post-infection (PI) using diminazine aceturate injection. Parasitemia
was detected on day 4 PI and fluctuating throughout the infection period. In the infected group, one
animal died unexpectedly on day 23 PI without showing the neurological symptoms of surra, while a
second animal was euthanized on day 15 post-treatment due to poor body condition induced by the
infection. Mean total erythrocyte counts (RBC), packed cell volume (PCV) and hemoglobin (Hb)
showed marked decline in the infected group on day 16 PI onwards. Mean corpuscular volume (MCV)
and mean corpuscular hemoglobin concentration (MCHC) remained normal. Thrombocyte counts in
the infected animals decreased dramatically below the control group and normal range on day 8 PI
onwards. A neutropenia accompanied with monocytosis was detected in the infected group on day 6
and 16 PI respectively, and fluctuated during the disease. In conclusion, the inoculum of 10 2
trypanosome/ kg body weight had established an acute disease distinguished by a normocytic,
normochromic anemia and one fatal outcome 23 day PI.
Blastocystis Symposia
BS1
Blastocystis since its discovery in the beginnings of 1900 have travelled through time totally enmeshed
in controversy and contradictions in all aspects of the organism including its phylogenetic status,
biochemical, biology, immunology and most importantly the pathogenic role it was supposed to exert.
The discussion traces its past, highlights major achievements in many of these fields and surface current
thinking especially in terms of its pathogenic role and the possible mechanisms involved in the process.
BS2
Blastocystis sp. is one of the most common intestinal parasites of humans and animals. Despite its
discovery in the early 1900s, many fundamental biological aspects of the parasite such as taxonomy,
pathogenicity and modes of transmission have just recently been revealed in the last decade with the
application of modern molecular tools. Phylogenetic analyses based upon small-subunit ribosomal RNA
gene, cytosolic-type 70-kDa heat shock protein, translation elongation factor 2 and noncatalytic B
subunit of vacuolar ATPase clearly demonstrated that Blastocystis is a stramenopile. Thus Blastocystis
is neither a fungus nor a protozoan. Blastocystis is placed in a newly created Class Blastocystea in
the Subphylum Opalinata, Infrakingdom Heterokonta, Subkingdom Chromobiota, Kingdom Chromista.
This classification makes Blastocystis the first chromist known to parasitize humans. The stramenopile
grouping of Blastocystis is rather unusual since the diverse group includes slime nets, water moulds
and brown algae. Blastocystis is most closely related to Proteromonas lacertae, a flagellate of the
hindgut of lizards and amphibian, although it does not posses any flagella. Based on the genetic distance
between homologous genes, Blastocystis sp. from humans and animals can be potentially divided into
12 or more species. Nevertheless the speciation of Blastocystis remains a great challenge mainly
due to its low host specificity. Studies on the association of a particular subtype with its pathogenic
potential have produced conflicting results but subtype 3 is the most frequently reported to be associated
with disease. Recent molecular studies have also provided evidence for the zoonotic and waterborne
transmission of Blastocystis sp.
BS3
Chandramathi S1, Suresh K1, Anita ZB2, Kok Hoe C3, Kuppusamy UR3 and Mahmood AA3
1Department of Parasitology
2Unitof Clinical Oncology
3Department of Molecular Medicine
Previous evidences have shown that Blastocystis hominis, one of the most widespread intestinal
protozoan parasites in humans, has a pathogenic role in causing intestinal inflammation. Studies have
reported that parasitic infections trigger inflammatory processes to produce free radicals (namely
reactive oxygen species or ROS) to kill the invading parasites. Excess of ROS can cause oxidative
stress, a contributory factor of cancer. However, association of B. hominis with cancer is yet to be
explored. Thus in present study, we aimed to investigate the effect of B. hominis on colorectal cancer.
The study involves 3 areas: assessment of oxidative stress level in A) cancer patients with and without
intestinal parasitic infection and B) in vivo model with B. hominis infection; C) in vitro study to
evaluate the effect B. hominis towards the growth of colorectal cancer cells. Results showed that
colorectal cancer patients (majority with B. hominis infection) exhibited high level of oxidative protein
damage. Non-cancerous individuals with intestinal parasitic infections (majority with asymptomatic B.
hominis infection) had high levels of oxidative damage to lipid and protein. Rats with asymptomatic
B. hominis infection exhibited high levels of oxidative damage to lipid and protein. The pro-inflammatory
cytokine levels in these rats were also elevated. These findings imply that B. hominis infection could
cause significant oxidative burst which may lead to inflammatory processes. Moreover, in vitro study
indicated that antigen from symptomatic isolate of B. hominis is more pathogenic compared to the
asymptomatic isolate as it caused higher levels of inflammatory reaction and proliferation rate in
colorectal cancer cells. In conclusion, the study has demonstrated that B. hominis can increase oxidative
damage and enhance the growth of colorectal cancer cells. Hence, it is very important to include
treatment procedures in patients with Blastocystosis.
BS4
Department of Parasitology, Faculty of Medicine, University of Malaya, Lembah Pantai, 50603 Kuala Lumpur,
Malaysia
Blastocystis sp. is a common intestinal parasite detected in many faecal surveys of humans and animals.
To date, at least 16 studies have implicated contaminated water as a source of Blastocystis infection.
Of which, two studies had successfully detected the parasite using polymerase chain reaction (PCR).
The potential of Blastocystis sp. in causing a major outbreak must not be undermined due to its tiny
size, resistance to chlorination and low infectious dose. The present paper will highlight some current
challenges that we faced in studying Blastocystis sp. in water sources. Despite the accumulating
evidence on its association with irritable bowel syndrome and urticaria, the pathogenicity of the parasite
is still being debated. This has somehow hindered researchers from putting much effort in searching
for the organism in water. Although the tiny and pleomorphic nature of the Blastocystis sp. faecal
cyst have been previously described, its morphological appearance in water sources is yet to be studied.
In addition, Blastocystis sp. has been confirmed to exhibit low host specificity, hence the cyst recovered
from water sources could potentially infect humans. Since World Health Organization has in 2004
endorsed the inclusion of the detection of Blastocystis sp. into World Health Organization Guidelines
for Drinking-water Quality (WHO GDWQ), there is an urgent need to devise an affordable, sensitive
and specific method to detect and enumerate Blastocystis sp. cyst in water sources. There should be
integrative efforts among public health officers, clinicians, laboratory scientists, policy makers and
communities to cooperatively meet the challenges of this potential public health threat.
OP1
Voon S1, Peters W2, Boklin C3, Divis PCS1, Ambu LN4, Nathan SKSS4 and Singh B1
1Malaria Research Centre, Faculty of Medicine and Health Sciences, UNIMAS, Jalan Tun Ahmad Zaidi Adruce,
93150 Kuching, Sarawak, Malaysia
2Castle Village, Berkhamsted, England
3 Sepilok Orangutan Rehabilitation Centre, Sandakan, Sabah, Malaysia
4Sabah Wildlife Department, Wisma MUIS, Kota Kinabalu, Sabah, Malaysia
By using morphological methods, two malaria parasites were previously reported to occur naturally in
orangutans, namely Plasmodium pitheci and P. silvaticum. Following analysis of the small subunit
ribosomal (SSU r) RNA genes of Plasmodium in orangutans from Kalimantan, Indonesia, the authors
of that study concluded that orangutans are infected with four species of Plasmodium, including P.
vivax and P. cynomolgi. However, the phylogenetic analysis of the DNA sequences was flawed and
no morphological descriptions were provided. In order to identify the different Plasmodium species
occurring in orangutans, a molecular and morphological approach was therefore initiated. A total of
116 blood samples were collected from 40 different orangutans (27 captives and 13 semi-captives /
wild) at the Sepilok Orangutan Rehabilitation Centre, Sabah. These samples were collected bi-monthly
from the captive orangutans, and on occasions that semi-captive/wild orangutans were ill or injured,
from March to November 2009. Screening for Plasmodium DNA using nested PCR assays indicated
that seven of the 27 (25.9%) captive orangutans were malaria-positive at least once during the sampling
period. Eight of the thirteen (61.5%) semi-captives/wild orangutans were malaria positive. The SSU
rRNA and cytochrome b genes were amplified, cloned and sequenced. Phylogenetic analysis indicated
that there are at least three, if not more, species of Plasmodium that infect orangutans. Thin blood
films from one orangutan showed malaria parasites that appeared to be similar to P. pitheci parasites.
However, molecular characterisation demonstrated mixed infections, thereby underscoring the
importance of molecular methods for correctly identifying species of Plasmodium.
OP2
Ng SL1, Norzana AG2, Anisah N3, Yusof S3, Noraina AR3 and Chua KH1
1Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia
2Department of Anatomy, Faculty of Medicine, Universiti Kebangsaan Malaysia
3Department of Parasitology and Medical Entomology, Faculty of Medicine, Universiti Kebangsaan Malaysia,
Acanthamoeba keratitis (AK) is a sight-threatening infection, where patients eyes were infected due
to physical contact with this free-living amoeba from the environment. In this study, 4 clinical strains
were isolated from corneal scraping of keratitis patients, while the environmental strains were isolated
from aquatic and soil sources of four recreational locations in Malaysia. Identification of Acanthamoeba
was based on Pussard and Pons classification of cyst morphology and phylogenetic analyses of the
partial 18S ribosomal DNA sequences. Result revealed that A. castellanii, A. culbertsoni, A.
polyphaga and A. lenticulata were detected within these samples. All clinical strains belonged to
T4 genotype and both T4 and T5 genotypes were found in the environmental strains. Both axenised
clinical and environmental strains were capable to lyse all the keratocytes after 24-hour co-incubation.
Based on the study, these environmental isolates are most likely to be pathogenic and could be the
potential risk factor of AK.
OP3
International Medical University, 126, Jalan Jalil Perkasa 19, Bukit Jalil, 57000 Kuala Lumpur
This study was carried to establish the DNA profile of Sarcocystis species found in wild rodents in
Peninsular Malaysia. One hundred and forty six rodents belonging to 7 species trapped in the states
of Johor, Selangor, Kelantan and Kedah were examined. Rodents as an intermediate host to Sarcocystis
pose a public health. Studies have shown the prevalence rate of Sarcocystis in Southeast Asia to be
high. Human infections with Sarcocystis spp. from rodents results in human muscular sarcocystosis,
implicated with myalgia, erythematous subcutaneous nodules, fever, bronchospasm, cough, headaches,
loss of appetite, weight loss and lethargy.
Hematoxylin and eosin (H&E) stained sections of the tissues from the wild rodents showed the
presence of Sarcocystis species. In the study using light microscopy, a total of 146 thigh muscles
were examined and 73 (50%) were found to be positive. Morphological observation showed that there
may be 3 different species infecting these rodents. The brain sections were found not to contain any
cysts. To identify the species present in these wild rodents, DNA extraction was carried out on paraffin
embedded blocks of tissue using 5 prime archive pure DNA cell/tissue kit. DNA profiling was done
for the identification of the different species. The results of the analysis will be reported at the
conference.
OP4
Aida Syafinaz M1, Jeffery J2, Noraishah Mydin AA2 and Tay ST1
1Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur,
Malaysia
2Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia
OP5
Distribution of insect species at high elevation becomes more important, given the presence of huge
numbers of high rise buildings in urban areas. Therefore, a study was conducted to investigate the
distribution of forensically important entomofauna at high rise building in Kuala Lumpur, Malaysia.
Two monkey carcasses were used. One was located at the top floor of Clinical Block of UKM Medical
Centre (UKMMC) and another one was located at the ground level at the backyard of UKMMC as
control. Both carcasses were monitored daily for 30 days. It was found that the decomposition process
was slower for the carcass located at the top floor (approximately 130 feet height). Numerous
entomofauna species were found on the carcass located on the ground, namely Ch. megacephala,
Ch. rufifacies, Ch. nigripes, Sarcophaga spp., Hemipyrellia sp., Hydrotea spp., Musca sorbens
and other insect species. For carcass located at the top floor, only three species of flies were found,
which were Synthesiomyia nudiseta, Sarcophaga sp. and Megaselia sp. The difference in
entomofauna distribution was most probably due to the ability of certain flies to reach high altitude
and could survive with different types of environmental conditions. The above findings highlighted the
importance of better understanding of fly behaviour and distribution in assisting forensic investigations,
especially when death occurs at high rise buildings.
OP6
Toxoplasma gondii (T. gondii) is one of the most well-known protozoan parasites causing a disease
called toxoplasmosis which infects many warm-blooded domestic and wild animals, including humans.
Due to a complex life cycle of T. gondii, these combined of sexual and asexual cycles thus create an
unusual population structure found in the host. Among the vast majority of T. gondii strains, the three
main clonal lineages belong to type I, II, and III are predominately found in Europe and North America.
In contrast, T. gondii isolates from human patients in South America represent polymorphic or divergent
lineages. Unfortunately, there was very limited data on the association between T. gondii strains and
human toxoplasmosis reported from Asia. It is postulated that infrequent sexual recombination,
population and geographical origin have been influenced these differences in T. gondii strains. In human
infections, type II strain is found in majority of cases which has thus far been reported in North America
and Europe. However, the links between Toxoplasma genotype and severity of disease is still a matter
of debate, particularly in patients with immunodeficiency. This review highlights the global distribution
of T. gondii genotypes in associating with human toxoplasmosis, recent development on different
methods of genotyping, and provides important implications on typing study in the management of human
toxoplasmosis in the near future.
OP7
A study was conducted to investigate the immunosuppressive effect of Trypanosoma evansi in goats
given Intranasal Pneumonic Mannheimia (IPM) vaccination. Twenty male goats were divided equally
into four groups. Groups 1 and 2 were inoculated intravenously with 104 trypanosomes / animal, while
goats in groups 3 and 4 served as uninfected vaccinated and uninfected unvaccinated controls,
respectively. Goats in group 2 were treated with diminazine aceturate two days before primary
vaccination. Groups 1, 2 and 3 received intranasal spray of 1 ml of Pneumonic Mannheimia (IPM)
vaccine on day 30 post infection (PI) and a booster dose on day 44 PI. On day 58 PI all goats were
inoculated intratracheally with 4 ml of live Mannheimia haemolytica organisms (106/ml) each. Blood
samples were collected weekly and analyzed for IgG levels using antibody-ELISA assay. All goats
were killed on day 72 PI, lung lavage fluid was collected and analyzed for IgA levels, while lung lesions
were assessed grossly. All goats became positive with T. evansi 4 to 6 days post infection. Group 1
remained positive throughout the experiment while Group 2 goats were negative 24 hours after
treatment until the end of the experiment. All the goats survived the experiment except for one in
group 3 which died before the challenge infection due to an unrelated cause. No significant difference
(P>0.05) was found in lung IgA levels nor lung lesion scores between the groups. Although serum
IgG levels between the four groups did not differ significantly, highest IgG level was detected in the
vaccinated group with the lowest level in the T. evansiinfected group. Thus the IgG findings imply
that T. evansi in the early stage of the infection compromised the immune response of intranasally
vaccinated goats.
OP8
Zuha RM1, Tasnim AR1, Zalifahtulhusna Z1, Khairul O1, Nazni WA2 and Baharudin O3
1Program Sains Forensik, Pusat Pengajian Diagnostik dan Kesihatan Gunaan, Fakulti Sains Kesihatan,
Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
2Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
3Jabatan Sains Bioperubatan, Pusat Pengajian Diagnostik dan Kesihatan Gunaan, Fakulti Sains Kesihatan,
Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
This research observed the developmental data of a cosmopolitan scuttle fly Megaselia scaris (Loew)
at selected laboratory temperature. M. scalaris was obtained from colony maintained in the Department
of Biomedical Science Insectary, Universiti Kebangsaan Malaysia. Rearing took place in a digital growth
chamber set at 27, 30 and 33C. Mean developmental time at 27C were: egg period, 15 hours (h);
feeding larva period, 58.2 2.5 h; postfeeding larva period, 23.0 1.8 h for male and 19.3 1.6 h for
female; pupa period, 179.7 6.9 h for male and 179.6 5.8 h for female. Developmental time at
30C were: egg period, 13.5 h; feeding larva period, 59.5 h; postfeeding larva period, 12 h for male
and 12.7 2.8 h for female; pupa period, 176.1 5.6 h for male and 175.3 5.7 h for female.
Developmental time at 33C were: egg period, 11.5 h; feeding larva period, 46.5 2.8h; postfeeding
larva period, 13.3 2.6 h for male and 24 h for female; pupa period, 176.3 7.8 h for male and 179.0
3.5 h for female. Total developmental time of M. scalaris from egg stage to adult emergence at
27C was approximately 275.9 h (11.4 days) for male and 272.1 h (11.3 days) for female; at 30C,
261.1 h (10.9 days) for male and 261 h (10.9 days) for female; and at 33C, 247.6 h (10.3 days) for
male and 261 h (10.9 days) for female.
OP9
Malaria is still a public health problem in Malaysia. Plasmodium falciparum has been characterized
based on their MSP genes in Southeast Asian countries. However, no much known about the population
structure of P. falciparum in peninsular Malaysia. MSP1 (block2) and MSP2 (block3) genes of P.
falciparum were genotyped from 36 samples collected from Pahang using nested PCR. The prevalence
of RO33 and K allelic families were 38.7% and 35.5%, respectively. MAD20 allelic family had the
lowest frequency (3.2%). Of the MSP2 allelic families, FC27 showed higher prevalence (12.9%)
compared to 3D7 (9.7%). The complexity of P. falciparum infection was 1.6. It would seem that
low complexity and distribution of the family alleles of MSP1 and MSP2 genes reflect the low intensity
of malaria transmission in peninsular Malaysia.
OP10
Blastocystis sp. is a common intestinal parasite found in faecal surveys. It affects 27 to 60% of people
in developing countries. Sporadic and scanty studies have implicated its waterborne potential. Hence,
it is important to mount a survey to access its occurrence in various water sources. This is essential
as World Health Organization has endorsed the inclusion of Blastocystis sp. detection into World Health
Organization Guidelines for Drinking-water Quality (WHO GDWQ). In this study, a total of 607 water
samples including drinking water (308), filtered water (14), lake (37), pond (5), river (208), tap (33)
and well (2) water with various volumes (0.5, 1, 5 and 10L) were collected from May 2008 to April
2010 in Peninsular Malaysia. Water samples were processed using centrifuge or flatbed membrane
filtration system in order to obtain water sediment, which was subsequently subjected to in vitro
cultivation. Blastocystis sp. was found in 13.8% (84/607) of the samples. From the samples detected
positive for Blastocystis sp., 98.8% (83/84) were river water samples and 1.2% (1/84) was lake sample.
The parasite was found in 39.9% (83/208) river water samples and 2.7% (1/37) lake sample but was
not found in all other water samples (drinking water, filtered water, pond, tap and well water). This
study suggests that river water could be the reservoir for waterborne transmission of Blastocystis
sp. In conclusion, our study has successfully provided evidence of the occurrence of Blastocystis sp.
in water sources. Hence, there is a need to design a systematic and affordable detection method to
enumerate Blastocystis sp. in water sources.
OP11
Tan Zi Ning1, Wong Weng Kin2, Rahmah Noordin2 and Lim Boon Huat1
1School of Health Sciences, Universiti Sains Malaysia, Malaysia
2Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Malaysia
Amoebic liver abscess (ALA) is the most common clinical manifestations of amoebiasis in human
caused by an enteric protozoan, Entamoeba histolytica. This neglected disease has claimed about
100,000 lives and inflicted many more annually. Indirect hemeagglutination assay (IHA), Cellognost
Amoebiasis (Dade Behring Marburg GmbH, Germany) is frequently used in diagnosis of ALA, but
it is costly and not suitable to be used in mass screening of invasive amoebiasis, particularly in endemic
developing nations. Thus, the objective of this study is to identify potential antigenic proteins from E.
histolytica crude soluble antigen (CSA) and ether extract antigens (EEA) for the diagnosis of ALA
by using experimentally infected hamster serum samples. Thirty Golden Syrian hamsters were each
inoculated with 1 x 106 of E. histolytica trophozoites to produce ALA. Each animal was sacrificed
with 3x overdose of pentobarbital and the cardiac-punctured blood sample was collected to obtain the
serum. The CSA was prepared via sonication of trophozoites while the EEA was prepared by solubilizing
the trophozoites using ether. Protein concentrations from both antigen preparations were then
determined using Bio-Rad Assay. Subsequently, Western blot analysis on both CSA and EEA based
on the hamster ALA serum samples revealed that the ~70kDa as a potentially important antigen for
diagnosis of hamster ALA. In conclusion, this antigenic protein set the stage for further study in the
diagnosis of human ALA.
OP12
Cystic echinococcosis (CE) is a zoonotic disease affecting mainly various species of livestock and
humans. It is caused by metacestodes of dog tapeworms of the Echinococcus granulosus complex.
Preliminary data about the prevalence of hydatidosis in different intermediate hosts in Sudan revealed
that the disease occurs in all domesticated intermediate host species (camel, cattle, sheep and goats)
with the highest infection rates in camels. Nevertheless, infection rates in humans are not very much
clear. Moreover, new epidemiological data suggest a major influence of the locally prevailing parasite
genotype/species. In this study, a previously described PCR system for species discrimination was
used and the partial cox1 and nad1 genes of the obtained samples (5) were sequenced. As the origin
of the patients is widely distributed over central, western and southern Sudan, the disease seems to
occur sporadically in a large part of the country. All of five cysts samples were E. canadensis G6. It
was suggested that this strain may have a lower pathogenicity to humans due to its sporadic occurrence.
This is especially so, as all epidemiological conditions for autochthonous transmission of CE are given:
In rural areas there are large numbers of dogs in and around villages, and infection can occur with
offal from slaughterhouses or during unsupervised home slaughtering. Nevertheless, even if the parasite
may have lower infectivity to humans, the infection can occasionally get established and progress to
clinical CE and all samples from the patients in our study were viable and contained protoscolices.
OP13
Malaria Research Centre, Faculty of Medicine and Health Sciences, UNIMAS, Jalan Tun Ahmad Zaidi Adruce,
93150 Kuching, Sarawak, Malaysia
Previous studies have shown that small mammals including bats and squirrels are natural hosts to two
genera of haemosporidian parasites, Hepatocystis and Plasmodium. In Peninsular Malaysia, two
Hepatocystis spp. and three Plasmodium spp. have been described in small mammals. A previous
study to identify haemosporidian parasites in small mammals from Sarawak indicated that fruit bats
and flying foxes were infected with Hepatocystis spp. The main objective of this study is to determine
whether malaria parasites exist in other small mammals in Sarawak by using molecular and
morphological methods. Forty-seven blood samples were collected from small mammals: 17 squirrels,
25 rats, 4 tree shrews and a mouse deer from four sampling sites in Sarawak. These samples were
examined by nested PCR assay and five plantain squirrels (Callosicurus notatus) were positive using
Hepatocystis and Plasmodium-specific primers. The small subunit ribosomal RNA (SSU rRNA) and
cytochrome b (cyt b) genes were amplified, cloned and sequenced. Phylogenetic analysis based on
the SSU rRNA genes showed the presence of two different clades, one closely related to Hepatocystis
spp. while the other was closely related to Plasmodium. The cyt b gene sequences formed two
monophyletic groups. Molecular data of SSU rRNA and cyt b gene sequences showed that
Hepatocystis spp. and Plasmodium spp. tend to be host specific. Blood parasites from plantain
squirrels were morphologically identical with Hepatocystis vassali malayensis from a plantain squirrel
in Peninsular Malaysia. In conclusion, plantain squirrels from Sarawak are infected with Hepatocystis
spp.
OP14
The protozoan Trichomonas vaginalis causes vaginitis, urethritis and cervicitis in human and it is
sexually transmitted. Nevertheless it is commonly asymptomatic in both men and women. The variable
degree of pathogenicity of the parasite could possible due to its phenotypic and genotypic variations
and it warrants further investigation. Hence this represents the first to establish the phenotypic and
genotypic differences between the parasite isolates obtained from symptomatic and asymptomatic
individuals. In this study, we obtained the parasites from 6 symptomatic patients and 4 asymptomatic
individuals. Symptomatic isolates exhibited higher growth rate and peak parasite count compared to
asymptomatic isolates in Hollander medium. We found that parasites in symptomatic isolates generally
greater in size and tend to cluster together and this is not observed in all asymptomatic isolates.
Symptomatic isolates consistently showed greater binding affinities to FITC-labelled Con A (Canavalia
ensiformis) and acridine orange which may indicate differences in surface carbohydrate content and
level of DNA activity respectively. DAPI staining exhibited multiple nuclei in a single cell of
symptomatic isolates. Scanning electron microscopy revealed rougher surface in symptomatic isolates.
Meanwhile transmission electron microscopy showed the abundance of perinuclear structure in
hydrogenosome. We observed the multiple nucleated formations and this confirms the occurrence of
multiple nuclear divisions in symptomatic isolates. Sequencing of the actin gene showed variations
between these two groups of isolates. This study provides conclusive evidence on the existence of
two different groups of T. vaginalis which differ both phenotypically and genotypically.
OP15
Teh CH1,2,3, Nazni WA1, Lee HL1, Fairuz A4, Tan SB1 and Mohd Sofian A2
Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Kuala Lumpur,
Malaysia
Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia
Method and Statistic Section, Institute for Public Health, Kuala Lumpur, Malaysia
4Bacteriolgy Unit, Infectious Disease Research Centre, Institute for Medical Research, Kuala Lumpur,
Malaysia
This study was conducted to determine the in vitro antibacterial activity of Lucilia cuprina larval
extract against seven selected diabetic wound pathogenic bacteria via four bioassays, namely, the
turbidometric (TB) assay, colony-forming unit (CFU) assay, agar well-diffusion assay and minimum
inhibitory concentration (MIC) assay as well as characterizing the physiochemical properties of L.
cuprina larval extract via TB assay. TB assays demonstrated that L. cuprina larval extract was
significantly potent against all bacteria tested (p<0.001). Additionally, CFU assay had affirmed that
the larval extract was bactericidal against Pseudomonas aeruginosa, Escherichia coli, Klebsiella
pneumonia, and to a lesser extent, against Staphylococcus epidermidis (p<0.001). On the other hand,
agar well-diffusion assay also revealed the apparent potency of larval extract against the gram-negative
bacteria, P. aeruginosa with formation of 19.6 1.06 mm (n=10) inhibition zones. Furthermore, MIC
assay had further substantiated the above results in which as little as 0.78 and 1.56 mg/ml larval extract
were able to inhibit 58.57 3.38% P. aeruginosa and 57.15 3.43% E. coli. Besides, physiochemical
characterization of the potency of L. cuprina larval extract via TB assays revealed that the freeze-
dried or reconstituted larval extract was highly robust and able to withstand long-term storage (13
months) at -70C as well as demonstrated high thermal and freeze-thaw stability (n=5) without
significant loss of antibacterial activity against all bacteria tested.
OP16
Background: At present, the treatment regimes of toxoplasmosis still fail to eliminate Toxoplasma gondii
from the infected host, due to low brain penetration. Spiramycin, a macrolide antibiotic (P-glycoprotein
substrate), tested effective against toxoplasmosis. Metronidazole, a nitro-imidazole antibiotic which
inhibits the substrates of P450 (CYP)3a4 & P-glycoprotein. Hence, we developed a HPLC method
for simultaneous analysis of spiramycin and metronidazole and to determine their interactions in plasma
and brain of mouse model. LC separation done with Phenomenex C-18 (15 cm x 3.8 mm, 5 m) column
with mobile phase of acetonitrile-phosphate buffer at pH 2.5, flow rate of 1 ml/min, 29C and detected
at 232 nm, in a gradient run. Validations were carried out based on inter- and intra-day variability,
linearity, precision and accuracy. In vivo drug-drug interaction was performed in Balb/c mice. Twelve
mice were dosed orally; four-metronidazole 500 mg/kg, four-spiramycin 400mg/kg and four-
metronidazole 500 mg/kg and spiramycin 400mg/kg (30 min after metronidazole). The animals were
euthanized 2h after the administration of spiramycin. Experiments were repeated by euthanizing the
mice at different time points; 0.5, 1.0, 2.0, 2.5, 4.0, 6.0, 8.0, 10.0 and 12.0 hours. Brain and blood
were obtained, processed and subjected to HPLC analysis. Results: Recovery was above 75%. Intra-
and interday variability was within 15%. Range of linearity: 0.2550.0 g/ml, mean r2 = 0.999. There
was no matrix interference and the LLOQ was 0.25 g/ml. In mice, the co-administration of spiramycin
and metronidazole showed a 2-fold increase in brain spiramycin concentration compared to spiramycin
alone. However theres no significant difference seen in the plasma spiramycin concentration.
Conclusions: An effective HPLC method to simultaneously analyze metronidazole and spiramycin was
developed. Metronidazole has enhanced the delivery of spiramycin into the brain, probably due to its
role as P-glycoprotein inhibitor. This may render clinical benefit in the treatment of chronic cerebral
toxoplasmosis.
Poster Presentation
PP1
Basir R1, Mohd Yusof MAA1, Fazalul Rahiman SS1, Chung WC1, Jabbarzare M1, Yam MF1,3,
Ismail S2 and Mahmud R3
1Pharmacology Unit, Department of Human anatomy, Faculty of Medicine & Health Sciences,
Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
2Centre for Drug Research, Universiti Sains Malaysia, 11800 Penang, Malaysia
3School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia
TREM-1 functions as an immune regulator and inducer of pro-inflammatory cytokines release and it
can amplify the inflammatory responses in many diseases. Since malaria infection is associated with
the elevation of proinflammtory cytokines which gave rise to severe immunopathological reactions,
we investigated whether modulating TREM-1 release pathway would have a positive impact on the
major pro-inflammatory cytokines release during the infection and the impact on parasitaemia
development. Plasmodium beghei infection in mice were used in this study. TREM-1 levels were
measured systemically using ELISA methods. Parasitaemia were monitored by means of thin blood
film stained with Leishman on microscope slide and viewed under light microscopy. TREM-1 release
pathway in malarial mice was modulated using recombinant TREM-1/Fc chimera that antagonizes its
action. Results showed that inhibition of TREM-1 caused significant decrease in the release of TNF,
IFN and IL-6 during the critical stage of the infection. The inhibition however resulted in a faster
development of the parasitaemia. This suggests that TREM-1 pathway is involved in triggering the
release of pro-inflammatory cytokines during malaria infection as its inhibition also resulted in the pro-
inflammatory cytokines being inhibited. The increase in parasitaemia development was however a
disadvantage to the earlier finding and may be due to the decrease concentration of TNF and IFN
that were thought to provide some inhibition on the parasite growth. Based on the current findings,
further investigation is needed to evaluate TREM-1 as a potential chemotherapeutic target for malaria
as the balance between the advantages and disvantages needs to be outweighed.
PP2
School of Medicine, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia
PP3
Wan Omar A1, Ngah Zasmy U1, Hairul Bazli S1, Rukman AH1, Roslaini AM1, Rayani M2 and
Hatam GR2
1Medical Parasitology Unit, Department of Microbiology and Parasitology, Faculty of Medicine and
Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor Darul Ehsan
2Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
A number of protective antigens have been identified in animal models utilizing several kinds of
adjuvants, most of which are toxic and cannot be applied in humans. Furthermore, the purified antigens
tend to lose immunogenecity. Therefore, an effective and safe adjuvant for human use is one of the
keys for successful malaria vaccines. The merozoite surface protein 1 (MSP1) is one of the leading
vaccine candidates at the erythrocytic stage. This molecule has been identified in almost all of the
Plasmodium sp that infects humans, simians and rodents. In this study we have also evaluated the
cross protective immunity of both the MSP1 Plasmodium falciparum and P. berghei MSP1. The
rPbMSP1 formulated in alum was found to be immunogenic which induced high levels of specific
anti-rPbMSP1 antibody. The IgG2a response predominated over that of IgG1 during the challenge
infection in the vaccinated mice. Mice vaccinated with rPbMSP1 in alum mounted significant protective
immunity against challenge infection (P<0.01). On day 121 after the booster, three out of ten mice
immunized with rPbMSP1 in PBS survived parasite infection (P < 0.05). In contrast, eight out of ten
mice and five out of ten survived respectively in those vaccinated with rPBMSP1 and rPFMSP1 in
alum (P < 0.01). Hence, obvious protective effects of MSP1 in alum immunization prevented death
from P. berghei lethal infection in mice (P < 0.01). There was significant cross-protection of rPfMSP1
against P.berghei challenge infection. In total these observations provide excellent basis for clinical
assessment of protective immunity of MSP1 in human subjects. This mouse model in this study is a
basis for screening vaccine candidate for the control of malaria which is estimated to be between
300 to 500 million clinical cases and 2.7 million deaths per year.
PP4
Khadri MS1, Nurulhusna AH1, Altymysheva N2, Mohd Noor I1, Norazlina AH1, Khairul AM1,
Syed Abd Rahman SM3 and Abdullah AG1
1Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur, Malaysia
2School of Diploma in Applied Parasitology and Entomology, Institute for Medical Research, Jalan Pahang,
Space spraying of pyrethroid formulations remains the main method used to control dengue vectors.
Normally, mosquito mortality after space-spraying was determined 24 hours post-treatment. However,
mosquito survival at extended post-treatment period and at different height needs to be re-assessed
due to discrepancies in test and operational use of these formulations. In this study, mortality of Aedes
was determined 24, 48, 72 and 96 hours at post treatment and at ground and at 1.5m height. A
formulation of 1.7% cyfluthrin (AEDIS ULV15) and 10.35% permethrin (AquaResigen) was
applied using an ULV portable generator against Aedes mosquito in a simulated field trial. Sucrose-
fed and caged Aedes aegypti and Aedes albopictus adults aged 5-7 days were held at 1.5 m height
and placed at 2 m intervals up to 14 m in a straight line at an open field together with MgO coated
slides for collection of insecticide droplets for analysis. The insecticide was space-sprayed against
the test mosquitoes according to standard WHO guidelines. The mortality of the mosquito was recorded
in laboratory at 24. 48, 72 and 96 hours post treatment. Results showed that low mortality and recovery
of mosquitoes were still occurring at 48, 72 and 96 hours post treatment. Approximately between 48
to 96 hours, 2-16% of Ae aegypti treated with cyfluthrin formulation whereas 1-24% of the mosquitoes
treated with permethrin formulation recovered. Similarly, high percentage of Ae albopictus recovered
especially those placed at the ground level. Hence these results indicated that not all knock down
mosquitoes died at 24 hours which may recover and survive. There is thus a need to re-assess the
post-treatment mortality determination.
PP5
Chloroquine (CQ) is considered as a first-line for prevention and treatment of malaria and is widely
used in Yemen. The chloroquine resistance (CQR) problem in Yemen is constantly growing since the
detection of indigenous cases of P. falciparum CQR in 1989. This study was carried out to determine
the prevalence of chloroquine resistance (CQR), based on pfcrt 76T mutation, in P. falciparum isolates
from Yemen. The pfcrt is the P. falciparum chloroquine resistance transporter, which is a digestive-
vacuole related transmembrane protein. Substitution of threonine (76T) for lysine (K76) at position 76
(K76T) in the pfcrt gene confers resistance of P. falciparum to chloroquine. The prevalence of pfcrt
76T mutation was high (66 of 81 isolates, i.e. 81.5%). High prevalence of pfcrt 76T was reported
among age group > 10 years. Almost two- third of the mutation was reported in male (63.6%) as
compared to female (36.4%). The prevalence in rural areas was higher than in urban areas. The
resistance was significantly associated with younger age (d 10 years), low household income and
not using bednets. Non-educated individuals and not working showed protective association (OR =
0.24; 95% C1 = 0.07-0.78; P = 0.016 and OR = 0.14; 95% C1 = 0.02-1.16; P = 0.034, respectively)
with resistance. By using logistic regression analyses, the age group d 10 years and the low household
income were significant with the CQR. This study revealed high prevalence of pfcrt 76T in the P.
falciparum isolates, thus suggesting that this gene marker will be most predictive for chloroquine
resistance in endemic areas. Surveys to determine the prevalence of pfcrt 76T will be useful in areas
that still use CQ.
PP6
Long-lasting factory-impregnated bed nets are increasingly used in malaria control programme as a
mean of personal protection against the anopheline vectors. Among these, two such nets, the
PERMANET and OLYSET net are used in several malaria endemic countries. In Laos, more than
50,000 nets were distributed and used extensively in several provinces. The long-term effectiveness
of these nets, however, needs to be evaluated. Fifty Permanet and 50 Olyset net were collected
randomly from the Loatian households after using the bed nets for two years. The nets were sent to
Institute for Medical Research (IMR), Kuala Lumpur for bioassay study. Each net sample on receipt
at IMR, was stored immediately at 4C until used. Standard WHO tube bioassays were conducted to
determine the effectiveness of these nets against Anopheles maculatus. The results indicated that
PermaNet mosquito net showed higher bioefficacy compared to Olyset mosquito net. The adult mean
mortality for PERMANET and OLYSET net was 99.17% and 81.07% respectively. There is no
significant difference in the bioefficacy among the Permanet (p>0.05; F= 0.67) and among Olyset
mosquito nets (p>0.05; F=1.97). However, highly significant difference was found in the bioefficacy
between the Permanet and Olyset mosquito nets (p<0.001: t=7.103). These results demonstrated the
long-lasting effects of these nets under field condition. However, there are many variables in the field
that may affect the performance of these nets.
PP7
Khadri MS1, Nurulhusna AH1, Altymysheva N2, Mohd Noor I1, Norazlina AH1, Khairul AM1,
Khatijah I3, Hanipa S3 and Abdullah AG1
1Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur, Malaysia
2School of Diploma in Applied Parasitology and Entomology, Institute for Medical Research, Jalan Pahang,
MOH Malaysia
A pyrethroid space spray formulation, AEDIS ULV15 containing 1.7% cyfluthrin, currently has been
utilized by some health authorities to control Aedes population. The objective of this study was to
determine the field effectiveness of AEDIS ULV15 compared to a 10.35% permethrin formulation
(AquaResigen) in controlling Aedes mosquito in a simulated field trial applied using an ULV generator
(Fontan). Sucrose-fed and caged Aedes aegypti and Aedes albopictus adults aged 5-7 days were
held at 1.5 m height and placed at 2 m intervals up to 14 m in a straight line at an open field together
with MgO coated slides for collection of insecticide droplets for analysis. The insecticide was space-
sprayed against the test mosquitoes according to standard WHO guidelines. The mosquito knockdown
rate was recorded at 20, 40 and 60 minutes post-treatment. Mortality of the mosquito was recorded
in laboratory 24 hours post-treatment. At 4, 6, and 8 meter distances, complete mortalities of both Ae
aegypti and Ae albopictus treated with permethrin were recorded, whereas complete mortality was
only observed in Ae aegypti treated with cyfluthrin at 6 m distance. Droplet analysis indicated presence
of ULV droplets and the ratio of VMD/NMD for cyfluthrin and permethrin formulations were 0.65
and 0.75, respectively, indicating fairly uniform distribution of droplet sizes. This limited simulated field
study indicated that the permethrin formulation was more effective against Aedes mosquito.
PP8
Deng Lu, Koou Sin Ying, Ng Lee Ching and Lam-Phua Sai Gek
Environmental Health Institute (EHI) of National Environment Agency, 11 Biopolis Way, #06-05/08,
Helios Block, Singapore 1386671
Vector-borne disease research often requires maintenance of laboratory colonies of vectors. Blood
feeding of mosquitoes is thus required for the maintenance of these colonies. Due to high cost and
inconvenience of using live animals as blood hosts, we developed a simple and user friendly blood
feeding system for artificial feeding the mosquitoes. The system consists of collagen membrane casing
and an in-house developed heating device. We carried out a parallel study between the membrane
blood feeding system and the guinea pig feeding method. Eight generations of Aedes aegypti and
three generations of Aedes albopictus were tested. Four parameters, namely, blood feeding rate,
fecundity, survival rate and hatchability were monitored. For Aedes aegypti, the results showed that
there is a significant difference in the feeding rate between the membrane feeding and the guinea pig
feeding methods (P = 0.012). However, there were no significant difference in the fecundity (P =0.556),
survival rate (P=0.715), and hatchability (P=0.932). Although, the feeding rate (85.3%) of membrane
feeding method is significantly lower compared to the rate (96.2%) of guinea pig feeding, the percentage
were at an acceptable level of 80% . For Aedes albopictus, the results showed there is no significant
difference between the two feeding methods (feeding rate P=0.068, fecundity P=0.887, survival rate
P=0.580 and hatchability P=0.564). Hence, we conclude that this collagen membrane blood feeding
system can be used for routine colonization of laboratory strains of Aedes aegypti and Aedes
albopictus.
PP9
Pang SC1, Lee ML1, Lam-Phua SG1, Chiang LP1, Deng L1, Maideen N2, Vythilingam I1
and Ng LC1
1National Environment Agency, Environmental Health Institute, 11 Biopolis Way, Helios Block, #06-05/08,
Singapore (138667)
2National Environment Agency, Environmental Health Department, Environment Building, 40 Scotts Road,
Entomological surveillance for malaria is challenging as the preferred habitats of Anopheles species
are frequently inaccessible. Vector investigation of malaria outbreak is crucial to pinpoint the right
Anopheles species so that appropriate vector control measures can be instituted to impede transmission.
In this study, we compared the species of mosquitoes between larval and adult surveillance. Larval
surveillance were performed within assessable sites from 9 am to 1pm while adult surveillance was
carried out using CDC-Light Traps (LT), BG-sentinel traps augmented with CO2, Human-Baited Net
Trap (HBNT) and Human Landing Catch (HLC) from 7 pm till 12am. Here we describe two case
studies: one in mainland Singapore, and the other on an off-shore island. On the mainland, larval surveys
in ponds with brackish water produced only 2 species Culex sitiens and Anopheles epiroticus.
However, more than 15 species were collected during adult surveillance around the breeding sites.
An. sinensis (>20%) was the major Anopheles species and surprisingly no An. epiroticus was
collected. On the off-shore island of Singapore, larval surveys yielded no An. epiroticus and 14% of
the larvae were An. barbirostris. However, An. epiroticus was the major Anopheles species collected
during the adult surveillance, constituting 19% of the total mosquitoes caught. HLC was the most
sensitive adult surveillance technique and LT was the best mechanical trap. This study shows that
entomological surveillance must incorporate larval and adult surveillance to enable a more holistic
understanding of the mosquito population on site.
PP10
The primary approach to control vector-borne diseases rely mainly on the application of insecticides.
The prolonged usage of insecticides may cause rapid development of insecticide resistance in many
insect species including mosquitoes. Aedes aegypti has been known to develop resistance to insecticides
in Singapore. The objective of the current study is to determine resistance/susceptibility status of Ae.
aegypti from historical and new sensitive dengue areas in Singapore. Biochemical studies of F1
mosquitoes reared from ovitrap collections were carried out using WHO microplate assays to
determine the enzymes activities in individual mosquitoes. The enzymes tested were
acetylcholinesterase (AchE), non-specific esterases (- and -esterases), glutathione S-transferases
(GST) and mixed-function oxidases (MFO). There were significantly increased (p<0.05) levels of
altered AchE in adult Ae. aegypti in all populations (n=170) but not in larval stage (n=223). Larvae
from all populations had elevated non-specific esterases activities. No evidence of - and -esterases
activities were found in adult mosquitoes. No significant (p>0.05) elevated GST activities were identified
in all populations except Clementi strain (new sensitive area). Only Ang Mo Kio strain (historical
sensitive area) showed elevated MFO enzyme levels (associated with pyrethroid resistance) in larval
stage. The results suggest that Ae. aegypti is developing lowered susceptibility to organophosphates.
Further investigation with bioassays and synergists studies are needed to provide direct evidence for
the mechanism involved.
PP11
Wan Najdah WMA1, Rohani A1, Zamree I1, Rahimi H2 and Lee HL1
1Medical Entomology Unit, Institute for Medical Research, Kuala Lumpur
2Pejabat Kesihatan Daerah Kuala Lipis, Pahang, Malaysia
Understanding local variability in malaria transmission risk is critically important when designing malaria
vector control programme. Using a combination of field data, satellite image analysis and GIS technique
we developed a map of malaria transmission risk area in Pos Lenjang. Entomological surveys which
consisted of adult collection and larval survey were conducted to determine the type of mosquitoes,
their characteristics and the abundance of habitats. Features like rivers, small streams, forest, vegetation
areas, roads and residential area were extracted from the satellite image. Base topographical maps
of the site were digitized and overlaid with entomological data. All digital data in the GIS were displayed
in the WGS 1984 coordinate system. The results showed that An. maculatus prefer to breed in clear
rock pool form on the bank of river and water falls. Map of villages with 400 m buffer zone visualizes
that more than 80% of An. maculatus immature habitats were found within the buffer zone. Changes
in breeding characteristics were also observed. In addition, adult population studies showed that a
total of 1,435 adult mosquitoes were collected by human landing catching technique from 17 mapped
villages. Anopheles maculatus Theobald (31.01%), Armigeres flavus Leicester (11.29%), Armigeres
annulitarsis Leicester (11.01%), Culex vishnui Theobald (9.55%) and Aedes albopictus Skuse
(7.04%) were the predominant species caught in the study area. As for larval survey studies, Anopheline
and Culicine larvae were collected and mapped from 79 and 67 breeding sites respectively. Satellite
imagery is a useful tool in mapping malaria risk areas and planning of an effective control programme.
PP12
Mark-Release-Recapture (MRR) experiments have been used for several decades to study longevity
and horizontal dispersal of mosquitoes. This paper compares field and laboratory strains of male Aedes
aegypti mosquitoes in terms of their dispersal and longevity. The study was conducted in Melaka
consisting predominantly of residential single-storey terraced houses. 4-6 old males were released and
BG-Sentinel adult traps were used to recapture them over a 7-day period. Flourescent powder was
used to mark the two strains with different colours. A total of 1,957 and 5,942 males were released
from field and laboratory strains respectively, and similar recapture rates (0.31% vs 0.32%) were
obtained. Dispersal was estimated in terms of median dispersal, mean distance travelled, and flight
range 90 (distance from the release point within which 90% of the released mosquitoes are expected
to be found). The field strain had higher values for these parameters (87m, 130m, 190m) compared
to the laboratory strain (22m, 54m, 115m) indicating that the former has better dispersal capacity.
Field strain also had higher daily survival probability (0.74) compared to laboratory strain (0.67), although
bootstrap analysis didnt reveal any significant difference between strains. Wing measurements showed
that the field strain males were bigger (although reared under same initial conditions) which could be
a likely explanation for its higher dispersal capacity. These MRR experiments give us valuable
information on male Aedes aegyptis behaviour in its natural habitat, which can be useful in setting
up Sterile Insect Technique (SIT) programmes as part of integrated vector control.
PP13
Rozilawati H1,3, Faudzi AY1, Siti Rahidah AA1, Nor Azlina AH1, Abdullah AG1, Amal NM1,
Wan Mansor H2, Hani H2, Apandi Y1, Noor Faezah1, Norziyana1, Nazni WA1, Zairi J3,
Yahaya MA3, Adnan CR3 and Lee HL1
1Institute
for Medical Research, Ministry of Health, Malaysia
2StateHealth Department, Kelantan, Malaysia
3School of Biological Sciences, Universiti Sains Malaysia, Pulau Pinang, Malaysia
Chikungunya infection has become a public health threat in Malaysia since the 2008 nationwide
outbreak. At the same time Aedes albopictus Skuse was identified as the chikungunya vector in the
state of Johor. In 2009, several outbreaks were reported in the state of Kelantan and entomological
studies were conducted in the state, in four districts, namely Jeli, Tumpat, Pasir Mas and Tanah Merah
in order to identify the vector responsible for the transmission of chikungunya. CHIKV case records
were obtained from the Kelantan State Health Department and the localities involved were identified.
Both larval and adult mosquitoes were sampled and transported back to the laboratory. The presence
of the virus was determined using reverse transcriptase PCR. A total of 1,245 mosquito larvae were
collected during the larval survey and 2,019 adult mosquitoes were collected using an aspirator. From
these collections, 640 mosquito pools were tested for the presence of CHIKV by RT-PCR. Ae
albopictus was the most abundant mosquito collected, followed by Culex sp., Armigeres sp. and
Anopheles sp. A total of 2, 814 artificial containers were inspected during the study. The House Index
(HI) in Jeli was 8.33%, Tanah Merah 6.45% and Pasir Mas 8.44%, container index (CI) was 33.33%,
7.44% and 10.61%, respectively, while the Breteau Index (BI) was 8.71, 8.39 and 8.44, respectively.
None of the mosquito samples were positive for the chikungunya virus; therefore the vector(s) of
chikungunya virus in the present localities could not be determined.
PP14
Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia,
43400 UPM Serdang, Selangor, Malaysia
Enterovirus 71 (EV71) is a member of the Picornaviridae family. It is one of the viruses which cause
hand, foot and mouth disease (HFMD). In recent years, several outbreaks of HFMD due to EV71
with severe neurological complications and significant mortality have been reported. Despite the serious
threat, no effective EV71 vaccine is available. Recently, we reported an induction of strong immune
response to a recombinant Newcastle disease virus (NDV) capsids displaying EV71 VP1 fragment
(NPt-VP11-100) in rabbit. The high immunogenicity suggests that this protein serves as a good candidate
for EV71 vaccine. However, since all EV71 animal models thus far are developed in mouse systems,
protective efficacy of this vaccine can only be tested in mice. Therefore, to investigate mice
immunogenicity to the NPt-VP11-100, we evaluated the type of immune responses developed by adult
Balb/c mice against the protein. NPt-VP11-100 protein induced high levels of anti-VP1 IgG production
in the mice. Purified VP1 antigen stimulated activation, proliferation and differentiation of splenocytes
harvested from the immunized mice. They also produced significant levels of Th1 cytokine, IFN-.
Taken together, NPt-VP11-100 protein is a potent immunogen in adult mice and our findings will provide
the data needed for testing of its protective efficacy in mouse models of EV71 infections.
PP15
Vinoth K1, Suresh K1, Nanthiney DR1, Umah RK2, Roslani AC3 and Kalyani R1
1Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur
2Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur
3Department of Surgery, University of Malaya Medical Centre, Lembah Pantai, 59100 Kuala Lumpur
Colorectal cancer (CRC) is a major cause of death worldwide. Although its incidence is higher in
developed countries, it is increasing rapidly in many parts of Asia, including Malaysia. A recent study
implicated Blastocystis hominis (B. hominis) in triggering CRC. B. hominis is commonly found in
human and animal stool samples, and is spread by water-borne transmission. In this study, colonic
lavage fluid from 30 CRC patients were collected and screened for B. hominis and other parasites.
The patients were asked to complete a questionnaire comprising various aspects to evaluate their
lifestyle. The samples were spun down and the sediments were cultured in Jones medium supplemented
with 10% heat-inactivated horse serum. They were then further incubated at 37C and screened for
B. hominis after 24 hours. DNA was also extracted from the sample for genotyping purposes. Formal-
ether concentration technique was carried out on the remaining sample to screen for other parasites.
All 30 samples cultured in Jones medium were negative for B. hominis. However, PCR amplification
with specific primers from the extracted DNA showed 3 samples positive for sub-type 3 (10%), 2
samples positive for sub-type 2 (6.7%) and 1 for sub-type 4 (3.3%). Modified trichrome stain (MTS)
showed 3 samples positive for Microsporidium sp. This study provides evidence that apart from stool
samples, colonic lavage could also be used for detecting B. hominis.
PP16
Malaysia.
Cancer patients undergoing chemotherapy are known to have immunosuppression that could trigger
latent parasitic infections in stools to emerge. However reports of such possibilities in cancer patients
are still scanty. This study aimed to investigate if intestinal parasites could emerge as opportunistic
infections in breast and colorectal cancer patients (n=46 and 15 respectively) by screening them
throughout the chemotherapy treatment regimes for 6 months. Breast cancer patients were on FEC
(5-Fluorouracil/Epirubicin/Cyclophosphamide) regime (comprising 6 chemotherapy cycles); colorectal
cancer patients were either on FOLFOX (Oxaliplatin/5-Fluorouracil/Folinic acid) or Mayo (5-
Fluorouracil/Folinic acid) regime (comprising 12 and 6 cycles respectively). Three urinary oxidative
indices were measured to study the treatment effect towards free radical damage. Based on the
questionnaires given the patients lifestyle, diet intake and home dwellings were unchanged throughout
the chemotherapy regime. Stool examinations of the cancer patients showed negative for any intestinal
parasitic infections prior to the chemotherapy. However, the patients were positive for Blastocystis
hominis and microsporidia infections during the intermediate chemotherapy cycles (cycle 2 - 4 out of
6 cycles and cycle 5 - 9 out of 12 cycles). This correlated with the increase of patients oxidative
indices level during the intermediate cycles suggesting that oxidative damage which is known to cause
immunosuppression may result in opportunistic infections. Therefore, cancer patients undergoing
chemotherapy should be screened repeatedly for intestinal parasites namely B. hominis and
microsporidia. This is because the infection by these parasites may intensify the free radical mediated
damage which could reduce the efficacy of chemotherapy treatments.
PP17
Blastocystis sp. is one of the most common intestinal parasites of human. This represents the first
study to determine the genetic diversity of Blastocystis sp. among a community of aborigine people
in Malaysia. A total of 58 out of 151 individuals (38.4%) were found positive for Blastocystis sp. by
in vitro cultivation. Blastocystis sp. isolates obtained were genotyped by PCR using seven pairs of
known sequenced-tagged site (STS) primers. Out of the 58 isolates, 41 were identified as one of the
known subtypes and mixed subtype infections were found in 15 (25.9%). Subtype 1 was most frequently
found, followed by subtype 3 and 5. The occurrence of high frequency of mixed subtype infections in
the community may indicate active transmission of Blastocystis sp. and hence the modes of the
transmission warrant further investigation.
PP18
Blastocystis sp., an intestinal parasite, has recently been implicated as a waterborne parasite. However,
there is a lack of protocol describing its recovery from water sources. Therefore, the main aim of
this study was to evaluate Blastocystis sp. cysts recovery using two methods, namely, flatbed membrane
filtration system and centrifugation. In flatbed membrane filtration system, water samples seeded with
a range of Blastocystis sp. cysts (2.2- 10.0 X 105 cysts) in 1L distilled water in a polyethylene beaker
was filtered at 250ml/min using flatbed membrane filtration system. Cysts were recovered by scrapping
the surface of membrane filter using 10ml phosphate buffered saline (PBS) as the eluting buffer followed
by centrifugation at 1,400xg for 10 min. In centrifugation method, water samples seeded with a range
of Blastocystis sp. cysts (1.4- 2.8 X 105 cysts) in 1L distilled water in a polyethylene beaker was
centrifuged at 1,400xg for 10 min to obtain cysts. All recovered cysts from two methods were
enumerated by trypan blue dye exclusion counts using a haemocytometer. Overall, recovery range
for flatbed membrane filtration was 63.0% 82.0% while centrifugation was 54.5% 63.6% with
mean recovery as 71.15.9% and 59.86.4%, respectively. Based on cost benefit analysis on both
the methods, we recommend that it is more cost effective to process small volume of water samples
(< 1L) using centrifugation while it is advisable to use flatbed membrane filtration for larger volume
of water samples (> 5L).
PP19
Nanthiney DR1, Suresh K 1, Mahadeva S2, Ho SH2, Kalyani R2 and Tan TC1
1Departmentof Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur
2EndoscopicUnit, 4th Floor, Menara Timur, University Malaya Medical Centre, Lembah Pantai,
59100 Kuala Lumpur
Blastocystis hominis (B. hominis) is one the most common intestinal parasites found in the intestinal
tract of humans and animals. Irritable bowel syndrome (IBS) is associated with changes in bowel
habits along with abdominal pain due to functional gastrointestinal disorder. The aim of this study is to
screen for B. hominis in colonic lavage collected from 30 confirmed IBS patients based on Rome
criteria III. The samples were collected from Endoscopic Unit, University Malaya Medical Centre.
The samples were spun down and the sediments were cultured in Jones medium supplemented with
10% heat-inactivated horse serum. They were then further incubated at 37C and screened for B.
hominis after 24 hours. DNA was also extracted from the sample for genotyping purposes. Formal-
ether concentration technique was carried out on the remaining sample to screen for other parasites.
All 30 samples were negative for B. hominis using direct microscopy and in vitro culture method. 5
IBS patient samples were positive for B. hominis. Out of 5 patients, 3 were Blastocystis sp. subtype
3 (6.7%) and 2 were Blastocystis sp. subtype 2 (6.7%) and 0 (0%) for subtype 4. This finding shows
that there is a need to screen for more colonic lavage samples from IBS patients to detect B. hominis.
PP20
The choice of preservatives, when collecting stools are important, when staining is considered of faecal
smears. The influence on staining of Blastocystis was assessed on parasites from stools spiked with
Blastocystis and preserved using 10% formalin, 70% alcohol and polyvinyl alcohol (PVA) respectively.
For each preservative fecal smears were made in triplicates and stained respectively with Fields,
Giemsa and Trichome stains. A human isolate of Blastocystis grown at 37C were used to spike human
stools. These spiked stools were kept at room temperature and 4C respectively to assess if temperature
can influence staining contrasts. Staining characteristics included morphology, clarity of nuclear and
cytoplasmic details, overall colour contrast and the ease to differentiate and detect Blastocystis from
faecal debris. The results showed that staining contrasts and clarity from stools persevered in 70%
alcohol were comparable with that of smears made from 10% formalin. Giemsa and Fields stains
showed similar contrasts, however Fields stain as previously reported, had shorter staining time.
Storage at 4C has been shown not to interfere with staining contrasts from both 10% formalin and
70% alcohol preservative. The study recommends that the 70% alcohol can be used as a preservative
and since the staining contrasts are comparable to that preserved in 10% formalin, it is suggested that
stools be collected in 70% alcohol as the preservative confers the advantage of exploiting the samples
for future molecular studies.
PP21
Blastocyctis hominis is an intestinal parasite that may cause abdominal pains and diarrhea. Infections
by B. hominis are usually treated with nitroimidazole drug which is metronidazole. However, there
are cases reported where B. hominis are resistant towards the drug. To date, research on the gene
responsible for the drug resistance in B. hominis has not been explored. Understanding the underlying
mechanism of this drug resistance may help in the treatment of infected patients. Reports have shown
that intestinal bacteria and protozoan parasites namely Helicobacter pylori, Giardia lamblia,
Entamoeba histolytica are treated with metronidazole. However, there have been reports on
resistance towards metronidazole in these microorganisms. Resistance of metronidazole in H. pylori,
G. lamblia and E. histolytica have been suggested to be triggered by the factors such as down
regulation of hydrogenosome or mitrochondrion like organelles (MLO), aerobic and anaerobic
environment, prolonged exposure to drug treatment, expression of specific genes, and mutation of
specific genes. It has also been reported that mutations in certain nitroreductase gene could be
responsible to confer resistance in H. pylori. Besides this, metronidazole resistance activity in G.
lamblia and E. histolytica are due to oxygen-insensitive nitroreductase (ntr) and nitroimidazole
reductase (NIMs) recombinant gene, and different level of NADH and NAD(P)H oxidase activity.
Investigation on resistance of B.hominis towards metronidazole is still lacking. All the above mentioned
evidences indicate that, there are possibilities for drug resistance in B.hominis to be similar with the
mechanism found in H. pylori and E. histolytica. This will be further discussed in the present study.
PP22
Apoptosis, a form of programmed cell death (PCD) in unicellular parasites have been previously
reported. Previous report showed human intestinal protozoan parasite Blastocystis hominis upon
exposure to metronidazole, have displayed morphological and biochemical features of PCD. The rate
of apoptosis in B. hominis in drug treated organisms however has not been established. This present
study demonstrates the rate of apoptosis of the B. hominis at different drug concentrations. Culture
tubes containing B. hominis were treated with 0.0001 and 1.0 mg/ml metronidazole respectively and
monitored for a period of 96 hr before introducing the respective viable parasites into drug-free fresh
growth medium. The results showed that the apoptosis rate was higher in Blastocystis treated with
0.0001mg/ml in contrast with 1.0mg/ml. However at the 48th hour of culture, the apoptosis rates in
culture tubes containing both the concentrations were reduced respectively. When the treated parasites
from both concentrations when harvested from 96 th hr of culture were introduced into fresh drug-
free medium, the number of apoptotic cells in both concentrations of drug increased respectively. In
conclusion, drug treated B. hominis when re-cultured in fresh medium showed higher rates of apoptosis.
Whether these drugs treated cells have gained resistance or have further changes at the biochemical
level remains uncertain. The study further implies the need to review the choice of drugs against
Blastocystis as drug treated organisms have been shown to influence viability and rate of apoptosis.
PP23
This present study examines the different biochemical properties found in trophozoites and pseudocysts
of Trichomonas vaginalis in asymptomatic and symptomatic isolates. Modified Fieldsstain, acridine
orange and DAPI staining were used to examine the distribution and intensity of nuclear material.
Nucleus in trophozoites from symptomatic isolates showed lesser staining intensity when stained with
both acridine orange and DAPI. The wall of trophozoites in symptomatic isolates stained less intensely
with FITC-ConA when compared to asymptomatic isolates suggesting a pathogenic role for these
lifecycle stages. However nucleus, in pseudocyst from symptomatic isolates showed brighter blue
and green fluorescence when stained with DAPI and acridine orange respectively when compared to
pseudocysts in asymptomatic isolates. DNA distribution and intensity also showed strong variation
between these forms. These differences could be correlated to the ultrastructual description. The study
confirms that phenotypic differences exist at the biochemical level between parasites from symptomatic
and asymptomatic patients.
PP24
Trichomonas vaginalis is a trichomonad species found in genitourinary tract of human which causes
trichomoniasis. Sexual transmission was considered as its mode of transmission. In the present study,
a total of 695 vaginal secretion samples from patients who came for routine Pap smear and sexually
active women with complaints of vaginal discharge were examined to detect T. vaginalis infection.
9% (n=467) of patients were asymptomatic and 32.81% (n=228) were symptomatic. 11 positive cases
(1.58%) of Trichomonas vaginalis were found and out of that 6 patients had cervical intraepithelial
neoplasia (CIN3) and 4 cases were asymptomatic cases. The prevalence rate of Trichomoniasis in
our study population is 1.58%. The 6 symptomatic isolates and 4 asymptomatic isolates were genotyped
using nested PCR method. Nested PCR amplified actin gene of T. vaginalis. Nested PCR offers a
sensitive and specific method for molecular typing of T. vaginalis. Further confirmation of sequencing
was obtained to show the genotypic differences between symptomatic and asymptomatic isolates.
Results indicated that there were genetic differences between symptomatic and asymptomatic isolates
of T. vaginalis. The present study suggested that genotypic differences revealed the pathogenic
variations potential infect human.
PP25
We obtained the flagellated protozoan Trichomonas vaginalis, which causes trichomoniasis from
vaginal swabs of symptomatic and asymptomatic female patients at a local clinic. Scanning electron
microscopy provided an opportunity to study the surfaces of three-dimensional images of trophozoites
and pseudocysts in asymptomatic and symptomatic isolates. The trophozoites of asymptomatic isolates
showed a smooth surface with few fine micropores as compared to the trophozoites from symptomatic
isolates which showed rough surface with numerous deep micropores. Numerous amoeboid forms
with many rough crater-like depressions were seen in trophozoites from symptomatic isolates. These
forms were found attached together implying that their surfaces were sticky. Pseudocyst from
asymptomatic isolates showed a less smooth surface as compared to the trophozites of the same without
any obvious noticeable micropores as compared to the pseudocyst from symptomatic which showed
rough surface, less micropores and attached to another implying that the surface is sticky. Pseudocyst
from asymptomatic isolates did not show any flagella, had smooth surface, with lesser surface crater-
like depression as compared to `pseudocysts from symptomatic isolates which showed more surface
crater-like depressions. Therefore, our results imply that phenotypic differences at the surface level
indicate that there were differences between trophozoites and pseudocysts of T. vaginalis in both
asymptomatic and symptomatic isolates.
PP26
PP27
Trichomonas vaginalis has been associated with cervical cancer. Therefore, the present study
investigated the differences between the effects of asymptomatic and symptomatic solubilised antigen
of T. vaginalis on the cell viability and proliferation of peripheral blood mononuclear cell (PBMCs),
human vaginal epithelial cell (HVEC) and cervical cancer CaSki cell (CaSki). We found that T.
vaginalis antigen from both asymptomatic and symptomatic isolates caused inhibition of PBMCs
indicating that the soluble antigens possessed the capability to suppress immune cell activity. On the
other hand, T. vaginalis antigen from-both the isolates induced the HVEC proliferation. This implicate
that T. vaginalis may own the ability to modify the normal growth of vaginal epithelial cells to become
hyper proliferative, which is the early step in carcinogenesis. Besides this, CaSki cells exposed to T.
vaginalis antigen showed a very mild proliferative activity (<10%) leads to a speculation that it may,
to a certain extent exacerbate the cervical cancer cell growth. However this will be further confirmed
and discussed by investigating the expressions of tumor promoting genes.
PP28
Mohammad R1,2, Ngah ZU1, Gholam Reza H2, Rukman AHR1 and Wan OA1
1Medical Parasitology and Entomology Unit, Department of Medical Microbiology and Parasitology,
Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Malaysia
2Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
I.R. Iran
PP29
Waterborne protozoan parasitic infections are the most frequently cause of morbidity in healthy
(immunocompetent) populations and may cause life-threatening diseases in immunocompromised and
immunodeficiency persons. The protozoan parasites are Acanthamoeba spp., Balantidium coli,
Blastocystis hominis, Cryptosporidium spp. Cyclospora cayetanensis, Entamoeba histolytica,
Giardia lamblia, Isospora belli, Naegleria fowleri, the microsporidia, and Toxoplasma gondii. These
protozoan infections remain common in both developing and developed countries, while, their
transmission is either via contact with recreational and surface water or consumption of contaminated
drinking water. Their potential for producing large numbers of transmissive and infective stages in
feces of infected peoples or animals constitutes persistent threats to both public and veterinary health
concerns. So far, more than 300 water-associated human outbreaks of protozoan parasitic diseases
have been well documented worldwide. The clinical scenarios from these outbreaks caused by these
pathogenic parasites varied from gastrointestinal, ocular or even fatal neurological involvements. In
recent years, there has been the increasing trend for the role that water can play in the transmission
of these pathogenic parasites to human. This review addresses the global epidemiological surveillances
of waterborne parasites, possible important mechanisms of transmission, advanced development on
detection methods of waterborne parasites for clinical laboratory purposes, the potential impacts of
parasites causing waterborne outbreaks diseases in human, and provides some insights into public health
policy making in safeguarding water quality.
PP30
Axenisation of Acanthamoeba spp. is aimed to obtain pure culture of trophozoites in order to study
its active form properties, especially when involving in vivo test and in vitro cell culture. Pure PYG
medium (0.75% peptone, 0.75% yeast extract and 1.5% glucose) is a common medium used for
axenisation and cultivation of Acanthamoeba trophozoites. However, this study showed that both clinical
and environmental isolates could only survive in pure PYG medium for 2 days after excystation. Optimal
axenisation medium (4% peptone, 4% yeast extract and 2% glucose) supplemented with 20% fetal
bovine serum (FBS) produced proliferative trophozoites. Stable axenised trophozoites were eventually
changed to cultivation medium (4% peptone, 4% yeast extract and 2% glucose) with 10% FBS and
pure PYG medium. Growth rates of both axenised clinical and environmental isolates between
cultivation medium with 10% FBS and 20% FBS were most likely to be the same. Only environmental
isolates could slowly adapt to pure PYG medium, while clinical isolates tend to encyst or die when
cultured in pure PYG medium. As a conclusion, cultivation medium with 10% FBS is more suitable to
maintain axenised Acanthamoeba trophozoites.
100 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 47th MSPTM Annual Conference
PP31
This study was conducted to determine the efficacy and the minimum cysticidal concentration (MCC)
of gentamicin sulphate and chlorhexidine digluconate, drugs commonly used in the treatment of infectious
keratitis against Acanthamoeba spp. A total of six isolates of Acanthamoeba (HUKM21, HSB1,
HUKM24, KRTG7, KRTG9 and KRTG10) were used. Doubling dilution of gentamicin starting from
40000g/ml g/ml and 200g/ml of chlorhexidine were performed in a microtiter plate using page
amoebic saline (PAS) to determine the MCC. After exposing Acanthamoeba cysts to antimicrobial
agents for 24 hours, the cysts were washed free of drugs and cultured cultured onto non-nutrient
agar overlaid with Escherichia coli. The growth of trophozoites was observed and recorded daily
for 14 days. Chlorhexidine successfully exhibit its cysticidal effects at therapeutic dose, however, it is
not so for gentamicin. The MCC for gentamicin sulphate range from 10000-20000 g/ml whereas the
MCC for chlorhexidine varied from 6.25g/ml to 50g/ml, much lower than the recommended
therapeutic dose. The MCC also differ according to the isolates used. Gentamicin sulphate has the
highest MCC and they exceed the recommended therapeutic dose for the treatment of keratitis. Based
on this study, chlorhexidine digluconate is the best antimicrobial agent against Acanthamoeba isolates
used, however, further investigations using more clinical isolates is needed in order to reaffirm the
result.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 101
47th MSPTM Annual Conference ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
PP32
Lim Boon Huat1, Tan Zi Ning1, Wong Weng Kin2, Foo Phiaw Chong1 and Rahmah Noordin2
1School of Health Sciences, Universiti Sains Malaysia, Malaysia
2Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Malaysia
Entamoeba histolytica is an enteric protozoan that causes intestinal amoebiasis, which may manifests
into the fatal amoebic liver abscess (ALA) if left untreated. Previous studies reported that membrane-
bound proteins of the parasite could be antigenic and play important role in diagnosis. However, some
other amoeba species might share similar membrane-bound proteins. Thus, to increase the specificity
of diagnosis, membrane-bound proteins can be removed by ether. The aim of this study is to compare
the protein profiles of crude soluble antigen (CSA) and ether extract antigen (EEA) of E. histolytica.
Firstly, 10 x 106 trophozoites were used to prepare either CSA or EEA. CSA was done by sonication
technique while EEA was prepared by adding ether to 1X PBS suspended trophozoites and the
supernatant was collected after removing the organic phase. Protein concentrations were determined
and both CSA and EEA protein profiles were compared by running 12% SDS-PAGE. Several protein
bands such as ~40kDa and ~32kDa not found in EEA protein profile were probably located in the
plasma membrane. It is interesting to note that both the antigen preparation techniques showed a few
prominent protein bands such as ~100kDa, ~45kDa and ~38kDa. Future study will focus on the
antigenicities of these proteins.
PP33
This study carried out to determine the sero-prevalence of Toxoplasma gondii antibodies in women.
A total of 240 samples of sera form the Obstetric qyna out patient clinic. The sero-positive were 131
out of 240 women (54.58 %). Among the age group 19-28 year the sero-positive 55 (48.25%) out of
114, comparing to the age group 29-38 year and age group 39-48 year were 62 (60.19%) out 103, 14
(60.87%) out of 23 respectively, but statisticaaly no significance according to age. Also found that
the women whom are seropositive and had no history of contact with cats were(54.58%), while women
whom are seropositive and have history of contact with cats were (57.4%), statistical no significance
difference between seropositive, factor contact with cat, and no difference according their residence
at different part of Tripoli-Libya.
102 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 47th MSPTM Annual Conference
PP34
Wan Omar A1, Ngah Zasmy U1, Hairul Bazli S1, Rukman AH1, Roslaini AM1, Rayani M2
and Hatam GR2
1Medical Parasitology Unit, Department of Microbiology and Parasitology, Faculty of Medicine and
Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor Darul Ehsan
2Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences,
Toxoplasma gondii infection is widespread in humans, although its prevalence varies widely from
place to place. Most infections in humans are asymptomaticbut the parasite can produce devastating
disease. In pregnancy,infection can result in congenital infection with severe sequelaeor late-onset
eye disease, and it is a frequent cause of encephalitisin severely immunosuppressed patients with AIDS.
A dipstick dye immunoassay was developed to detect immunoglobulinG (IgG) or IgM antibodies of
toxoplasmosis infection in humans. The assays employ peroxidase conjugatedto sheep anti-human IgG
and rabbit anti-human IgM as the visualizingagents and recombinant surface antigen of Toxoplasma
gondii (r SAG1) a soluble antigen of tachyzoites of Toxoplasma gondii as the detective antigen. The
antigen was transblotted on a nitrocellulose membrane. The assays are rapid (the whole test can be
completed within15 min), simple, and cheap, and they dont require any equipment. They are sensitive
and specific for the detection of anti-ToxoplasmaIgG or IgM antibodies and generally agree closely
with the resultsfrom the enzyme-linked immunosorbent assay. The assays are especiallysuitable for
field applications.
PP35
Aleuroglyphus ovatus is a common pest of stored products and is commonly found in store houses
and barn dust. A. ovatus may contaminate the processed food such as flour and cereals also and is
reported as one of the important causes for the development of occupational allergic diseases in
agriculture and baking industry. In this study, monoclonal antibody (Ao3B8) against A. ovatus was
produced using hybridoma technology. Low cross-reactivities of this antibody against different species
of bacteria, fungi, cockroach, fly and tick were detected. Sandwich ELISA was developed using this
antibody for the detection of A. ovatus in dust samples (20) randomly collected from different locations
of a university. The lowest detection limit of this assay was 0.001 g of A. ovatus. A. ovatus was
mainly found in the dust collected from an animal holding facility and some secluded areas of the
laboratories. In conclusion, this monoclonal antibody can be used for the detection of A. ovatus in
environmental dust samples.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 103
47th MSPTM Annual Conference ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
PP36
SRAS Berhad, No. 1 Jalan P2/19, Seksyen 2, Bandar Teknologi Kajang, 43500 Semenyih,
Selangor Darul Ehsan, Malaysia
Environmental monitoring at industrial set ups is crucial as it ensure public safety and prevents workers
from exposure to hazardous pollutants. At the present study, two methods were conducted to assess
the indoor environmental parameters. One being gas chromatography testing on air sample collected
by an air sampler on site by a NIOSH certified assessor. The other method is by employing an online
monitoring system built on SCADA platform to collect real-time data. A winery in the Klang Valley
was selected for the study. As fermentation is on-going in the production room, the emission of carbon
dioxide and volatile organic compounds were captured and analysed. Seven types of volatile organic
compounds were detected by gas chromatography, ranged from 0.068 mg/m3 to 2.895 mg/m3. On
the other hand, real-time monitoring system recorded total volatile organic compounds ranging from
0.03ppm to 28.29ppm. We found that standard assessment program provides a general view on
chemicals being emitted in the production floor and minimizes assessment cost, while real-time
continuous monitoring system is more efficient in picturing uncertainty thus offering better reliability
in pollution control. Volatile organic compounds and chemical gases usually present dynamically in the
ambient. An environmental monitoring framework at implementation stage has been developed. This
system is capable in achieving a cost effective long-term monitoring program, with the flexibility to
counter on-course any variations in the surveillance environment. The system will further offer a
dynamic system to be utilized in environmental impact studies and risk assessment, as well as decision
making in the short-run, policymaking on the long-run. The tested system has been incorporated into
a wide area network, thus offering the potential of better predictive ability and greater warning lead-
time for alarming conditions than that of separated, stand-alone surveillance modalities.
104 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 47th MSPTM Annual Conference
PP37
Eimeria tenella is an apicomplexan parasite that causes the economically important disease known
as coccidiosis in chickens. The spread of drug-resistance strains requires the development of new
drug targets, and this process can be facilitated by understanding the function of novel molecules in
the parasite. Recent studies have shown that the porin protein is essential in eukaryotic organisms.
Hence, this study was carried out to generate and characterise the porin cDNA sequence in E. tenella.
Plasmid DNA that contained the E. tenella porin gene (Etporin) cDNA was subjected to DNA
sequencing using the primer walking strategy and sequences generated were quality- and vector-clipped
using Phred and VectorTrim respectively. Assembly of the sequences using Bioedit resulted in a contig
of 1368bp in size. Analysis with ORF Finder shows that the most probable open reading frame is
879bp in length and codes for 292 amino acids. Blastx analysis against the GenBank database shows
that Etporin is 47% similar to the putative porin sequence of Toxoplasma gondii, while a search against
the Conserved Domain Database (CDD) shows that Etporin contains the Porin3 superfamily domain.
In addition, ClustalW alignment with porin sequences from various eukaryotic organisms shows that
the conserved VKXKX and GLK/ STK motifs are present in Etporin. Structure analysis predicts
that Etporin is composed of 19 -strands and adopts a -barrel architecture that is common to all
porin proteins. Taken together, the results of this study indicate that Etporin codes for the porin protein
in E. tenella.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 105
47th MSPTM Annual Conference ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
PP38
Yao PP1, Tay YL1, Hasidah MS1, Embi MN1 and Wan KL1
1School of Biosciences and Biotechnology, Faculty of Science and Technology,
Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor
Eimeria tenella is the most pathogenic of the intracellular protozoan parasite Eimeria spp. that causes
coccidiosis in chickens. The economic impact of coccidiosis to the poultry industry and concerns relating
to drug-resistance of the parasite towards current chemotherapeutics strengthen the need for novel
drug targets. Glycogen synthase kinase-3 (GSK-3), a multifunctional serine/threonine protein kinase
involved in the regulation of diverse cellular processes, represents a suitable candidate. In this study,
the in silico identification and characterisation of the putative E. tenella GSK-3 (EtGSK-3) gene
was carried out. Expressed sequence tags (ESTs) corresponding to GSK-3 were compiled and mapped
to the E. tenella genome sequence. The structure of EtGSK-3 was predicted using TWINSCAN
and refined with RNA-seq data. The resulting EtGSK-3 gene model is 3119bp in length with an open
reading frame (ORF) of 1239bp that codes for 417 amino acids. Blastx analysis of the predicted ORF
revealed 84% similarity to Toxoplasma gondii protein kinase 3 and 80% similarity to Plasmodium
falciparum GSK-3. ClustalW alignment with GSK-3 sequences of other eukaryotes showed that
EtGSK-3 contained the 11 conserved subdomains essential for protein kinase activity. Results of this
study indicate that EtGSK-3 codes for the GSK-3 protein in E. tenella.
PP39
Faculty of Medicine, Universiti Teknologi MARA (UiTM), Shah Alam, Selangor, Malaysia
Sarcocystis sp. infection was investigated in 20 necropsied captive wild mammals and 20 birds in
two petting zoos in Malaysia.The gross post mortem lesions in mammals showed marbling of the liver
with uniform congestion of the intestine and for birds there was atrophy of the sternal muscles with
haemorrhage and oedema of the lungs in two birds. Naked eye examination was used for detection
of macroscopic sarcocysts, and muscle squash for microscopic type.Only microscopically visible cysts
were detected in eight animals and species identification was not possible. Histological examination
of the sections of infected skeletal muscles showed more than 5 sarcocysts in each specimen. No
leukocytic infiltration was seen in affected organs. The shapes of the cysts were elongated, circular
and the mean size reached 254 x 24.5 m and the thickness of the wall up to 2.5m.Two stages were
recognized in the cysts, the peripheral metrocytes and large numbers of crescent shaped merozoites.
Out of 40 animals examined, 3 mammals and 5 birds were positive (20%). The infection rate was
15% and 25% in mammals and birds respectively. Regarding the organs, the infection rate was 50%
in the skeletal muscles followed by tongue and heart (37.5%), diaphragm (25%), and esophagus
(12.5%). Further ultra-structural studies are required to identify the species of Sarcocystis that infect
captive wild animals and their possible role in zoonosis.
106 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 47th MSPTM Annual Conference
PP40
Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur
A survey of wild rodent population of coastal and island of Peninsular Malaysia was carried out from
the months of May to August to determine their diversity and distribution of ectoparasites and
endoparasites. A total of 190 rodents were captured comprising of 4 areas: Kuantan (54.2%), Malacca
(20.5%), representing coastal and Penang (13.2%) and Carey Island (12.1%) representing island sites.
The most dominant species in Kuantan were Rattus rattus diardii (62.1%), followed by Rattus
norvegicus (34.0%), Rattus exulans (2.9%) and Rattus tiomanicus (1.0%) while in Carey Island,
only two species were captured; Rattus rattus diardii (56.6%) and Rattus tiomanicus (43.5%). All
rodents in Penang and Malacca captured were Rattus norvegicus (100%). The population showed
higher number of females (56.8%) compared to males (43.2%) from all sites. The ectoparasites
recovered fell under 3 broad groups, namely lice (Polyplax spinulosa), mites (Laelaps nuttali and
Laelaps echidninus) and flea (Xenopsylla cheopis) while endoparasites recovered were Cestodes
(Taenia taeniaformis, Rodentolepis nana and Hymenolepis diminuta) and Nematodes
(Nippostrongylus brasiliensis, Angiostrongylus malaysiensis, and Capillaria hepatica). Two main
factors were investigated to determine infestation in the rodent population, i.e. age-related and sex.
The rat populations from both habitats were observed to harbor more than one group of parasite. Of
all ectoparasites recovered, Xenopsylla cheopis is a known vector of murine typus and plague, posing
a potential health risk to humans.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 107
47th MSPTM Annual Conference ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
PP41
Norhidayu S1, Mohd Zain SN1, Md Suhaimi MFA1, Zakaria N 1 and JW Lewis2
1Institute
of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, 50603, Malaysia
2Schoolof Biological Sciences, Royal Holloway, University of London, Egham Hill, Egham, TW20 OEX, Surrey,
United Kingdom
The occurrence of macroparasites was studied from 543 stray cats in four urban cities from the west
(Kuala Lumpur), east (Kuantan), north (Georgetown) and south (Malacca) of Peninsular Malaysia
during January 2009 to August 2010. Five ectoparasite species were recovered namely,
Ctenocephalides felis, Felicola subrostrata, Haemaphysalis bispinosa, Heterodoxus spiniger and
Lynxacarus sp. with an overall prevalence of 46.2% Haemaphysalis sp. was found in all cities except
for Kuantan whereas Lynxacarus sp. was only found in Kuala Lumpur and Georgetown. Two cats
from Georgetown were infested with the dog louse, Heterodoxus spiniger and this represented the
first host record of this species in Malaysia. Up to nine species of helminths were recovered, with
overall high prevalences of infections of 83.0% in Kuantan followed by 75.1% in Kuala Lumpur 71.6%
in Georgetown and 68.0% in Malacca. The nine helminths comprised six nematode species (Toxocara
malaysiensis, Toxocara cati, Ancylostoma braziliensis, Ancylostoma ceylanicum, Strongyloides
sp., Physaloptera praeputialis) two cestode species (Taenia taeniaeformis, Dipylidium caninum)
and one trematode species (Platynosomum fastosum). Most helminth parasites were present in all
study sites except for the sole presence of Strongyloides sp. and the absence of Physaloptera
praeputialis in Kuala Lumpur. Levels of infection and infestation were also shown to be influenced
by host age and gender and the extent of parasite species richness in these four urban cities is
discussed.
PP42
A louse species, Heterodoxus spiniger, commonly found on dogs, is recorded for the first time in
cats from Peninsular Malaysia. A total 102 stray cats were examined from Georgetown, Penang in
association with the Society for the Prevention of Cruelty to Animal (SPCA).Two cats were infested
including a juvenile male and female, which were both found in close contact with each other prior to
capture. The number of lice ranged from up to 5 and 14 in male and female cats respectively. Other
ectoparasites recovered included the common cat flea, Ctenocephalides felis, although infestations
with lice and fleas did not appear to adversely affect the health of the stray cats.
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PP43
Acarology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur
Objectives: (a) To establish a Polymerase Chain Reaction (PCR) technique based on mitochondria
DNA (mtDNA) cyt b gene, for blood meal identification; (b) To compile data for mitochondria DNA
(mtDNA) cyt b gene sequence of some vertebrate animals in Malaysia; and (c) To deposit them in
international databases. Materials and Methods: The PCR technique was established based on
published information and validated using blood sample of four laboratory animals (guinea pig, rabbit,
SD rat and Balb/c mice) of which their whole gene sequences are available in GenBank. PCR was
next performed to compile gene sequences of nine different species of wild rodents. The species of
rodents were Rattus (rattus) tanezumi, Rattus tiomanicus, Leopoldamys sabanus, Tupaia glis,
Tupaia minor, Niviventor cremoriventor, Sundamys muelleri, Rattus rajah and Maxomys
whiteheadi. These rodents were caught in 8 different localities in Peninsular Malaysia. The primers
used for PCR are complementary to the conserved region of the cyt b gene of vertebrates mtDNA.
They are derived from the primers L14841 and H15149 described by Kocher et al (1989). A total of
88 blood samples were analyzed and 31 PCR products were sequenced. The sequences obtained were
then compared using a BLAST program to identify sequences with the most similarities. Results:
Seven new gene sequences have been deposited in GenBank database since September 2010 with
accession number HQ166262, HQ166263, HQ180173, HQ288326, HQ288327, HQ656820 and
HQ656821.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 109
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PP44
Tan Swee Beng1, Rohani Ahmad1, R Shamilah R Hisam2, See Kah Heng3, Zamree Ismail1,
Lokman Hakim Sulaiman4 and Lee Han Lim1
1Medical Entomology Unit, Infectious Disease Research Centre, Institute for Medical Research, Kuala Lumpur
2Medical Parasitology Unit, Infectious Disease Research Centre, Institute for Medical Research, Kuala Lumpur
3School of Applied Parasitology & Entomology (DAP&E), Institute for Medical Research, Kuala Lumpur
4Disease Control Division, Ministry of Health, Malaysia
Lymphatic filariasis still remains endemic in several states of Malaysia even though the control and
elimination programme for lymphatic filariasis was introduced since 1960. This study was conducted
to determine the possibility of local transmission of microfilariae parasites in field-collected mosquitoes
using the duplex polymerase chain reaction method in order to comply with the Global Elimination of
Lymphatic Filariasis by the year 2013. Adult mosquitoes from the endemic states of Johore, Kedah,
Kelantan, Pahang, Perak, Sabah, Sarawak, and Terengganu were collected by using the bare leg catch
(BLC) method from 7pm to 12pm and CDC light trap from 7pm to 6am commencing from the month
of February to October, 2010. A total of 232 pools of mosquitoes were obtained from a total collection
of 3501 mosquitoes comprising 23 species from six genera. DNA extractions were performed by using
Qiagen QIAamp DNA Mini Kit followed by DNA amplification and gel electrophoresis. The PCR
results indicated that no microfilariae parasites were detected in the mosquito. Based on these results,
local transmission of filarial worms was not detected.
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PP45
Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia
A survey was conducted to determine the current prevalence, polyparasitism rate and burden of soil
transmitted helminthiases (STH) among aboriginal school children in Satak, Raub district, Pahang state.
Malaysia. Two hundred and forty six (246) school children (119 males and 127 females) participated
in this survey. The result shows that, 93.5% of the children were infected with atleast one helminth
parasite species. Trichuris trichiura is the most prevalent, afflicting 83.7% of the subjects, it is followed
by Ascaris lumbricoides with 46.3% and the least is hookworm with 5.0%. Of the infected subjects,
42.3% were co-infected with atleast two helminthes, 51.2% had single infections while 6.5% were
un-infected. According to sex, 95% of all the males and 92.1% of the females were positive for atleast
one parasite. Age wise, 95.7% of the sampled children e10years and 91.6% of the children <10years,
were positive for helminthes. However, differences in the level of infection according to sex and age
groups were not significant (P>0.05). Worm burden analysis shows that, 14.1% of the infections by
A. lumbricoides and T.trichiura were heavy, 39.9% were moderate and 46%were light. All hookworm
infections were light. The survey confirmed that, STH infections are still a matter of public health
concern among the Malaysian aborigines. Polyparasitism and burden of infections among the subjects
could be due to the exposure to related risk factors. Efforts to improve their quality of life should
include improvement in sanitary facilities, proper disposal of human faeces, supply of portable water,
health education and introduction of school based helminth control programmes.
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PP46
Institute of Biological Sciences, Faculty of Science, University of Malaya 50603, Kuala Lumpur, Malaysia
Soil contamination with geohelminth eggs is a potential source of infection to human thus, poses a
threat to the public especially to children. Parasites commonly found in soil include Ascaris spp.,
Toxocara spp., Trichuris spp and Ancylostoma spp. Many reports have highlighted the geohelminth
impact on human worldwide in relation to diversity of parasite infection and prevalence, however, very
little is known of the status here in Malaysia. This study was conducted to determine the distribution
and biodiversity of soil-transmitted helminthiasis in 3 localities i.e. west (Kuala Lumpur), northern
(Georgetown) and southern (Malacca) states of Peninsular Malaysia, in particular the contamination
of sandpits surrounding childrens playgrounds. To date, a total of 200 soil samples were taken from
the sandpits of 40 playgrounds between October 2009 and August 2010. This study employed the
centrifugal floatation technique with the use of saturated NaCl (SGI.25) to determine egg counts
(E.P.G). Among the sites studied, all 40 playgrounds were contaminated with STH eggs namely;
Toxocara spp. Ascaris spp., Ancylostoma spp. and Trichuris spp. In fact, the prevalence of STH
eggs from sandpit of 40 playgrounds (200 samples) in 3 localities examined areas was 100%
respectively. Prevalence of Toxocara spp. (856.4 1.0546) was observed to be highest followed
with Ascaris spp. (740.8 2.5483), Ancylostoma spp. (388.4 1.2066) and Trichuris spp. (195.4
0.2256). The worst contaminated playground sites were in Penang (E.P.G level for Toxocara spp.
ranged from: 214.8-856.4; Ascaris spp.; 260.0-740.8; Trichuris spp.; 49.6-195.4; Ancylostoma spp.;
149.2-388.4).
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PP47
We report a series of dermatitis cases caused by the staphilinid beetles, Paederus fuscipes Curtis
among universitys students who stay in the residential colleges in Puncak Alam, Selangor, Malaysia
from January to December 2010. There were 12 hostels within the campus which hosted around 6,000
students. A total of 360 cases (6.0%) were recorded in the hostels Health Center throughout the
year and majority of the patients were came from the hostel which is located near to an oil palm
plantation. The age of students were ranged from 18-20 and all of them showed erythematous, edema,
vesicular papules, dermatitis linearis, painful blisters, burning sensation, pruritus (when sweating),
prolonged pigmentation and peeling skin after accidental crushing of the beetles. The common sites
of involvement were face, neck, shoulder and arms while other parts of body such as periorbital region,
chest, abdomen and legs were rarely seen. Most students noticed the symptoms in the early morning
after wake up. The patients were treated with fucidin cream and the symptoms resolved after 5 days.
These beetles are nocturnally active and enter the room whenever the light source is available. The
unintentional crushing of these beetles during sleep causes the release of its hemolymph (pederin)
which is the culprit for the dermatitis. Installation of window mesh, insecticide fogging, setting up physical
traps and increase the public awareness may decrease the incidence of paederus dermatitis. Several
factors attributed to this endemic dermatitis will be discussed.
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PP48
EFFECT OF BEEF LIVER, MEAT AND MIXED NUTRIENT AGAR DIETS ON THE
DEVELOPMENT OF SCUTTLE FLY, MEGASELIA SCALARIS (LOEW) (DIPTERA:
PHORIDAE)
Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
3Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia
This research investigated the effect of different types of diets on the development of cosmopolitan
scuttle fly, Megaselia scalaris (Loew) (Diptera: Phoridae). Diets used in this study were fresh beef
liver, beef meat, beef liver agar and beef meat agar. Larvae were cultured in beef meat recorded the
largest mean length, 4.21 2.02 mm, followed by beef liver agar (3.93 1.82 mm), beef liver (3.82
1.98 mm) and beef meat agar (3.81 1.77 mm). Highest mean weight was recorded in beef liver
and its agar form, 3.50 1.20 mg and 3.50 0.90 mg, followed by beef meat 3.40 1.20 mg and its
agar form (3.10 1.30 mg). Maximum mean length for M. scalaris pupa was recorded in beef liver
agar (3.47 0.80 mm), followed by beef meat agar (3.38 0.95 mm), beef liver (3.30 1.02 mm)
and beef meat (3.29 0.87 mm). Highest mean weight for pupa was recorded in beef liver and beef
meat agar, 3.50 1.10 mg and 3.50 1.30 mg respectively, followed by beef meat (3.10 1.40 mg)
and beef liver (2.70 1.40 mg). Nutrient agar recorded the lowest value of mean larval length (3.20
1.12 mm), pupa length (2.87 0.32 mm), larval weight (1.00 0.70 mg) and pupa weight (1.80
0.10 mg). Beef liver agar was also found to be an efficient alternative form of diet for culturing M.
scalaris larvae in laboratory because it gave optimum larval growth, odorless, clean and had a
consistent food texture.
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PP49
Department of Biomedical Sciences, Faculty of Allied Health Science, Universiti Kebangsaan Malaysia,
Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur.
A study on diversity and successional patterns of insect fauna on decaying animal carcasses was
conducted from 27th September to 28th October 2010 in a secondary forest at Universiti Kebangsaan
Malaysia, Bangi, Selangor. Collections of insects were made on nine animal carcasses which consisted
of three laboratory rats (Sprague Dawley, mean weight 0.508 kg), three rabbits (New Zealand White,
mean weight 2.538 kg) and three monkeys (Macaque fascicularis, mean weight 5.750 kg). A total
of 31088 individuals which comprised of three orders and twenty-two families were collected from
all carcasses. Insects belong to Formicidae family were the most abundant taxa which comprised of
more than 80% of total collected insects in all carcasses and they colonized carcasses from fresh to
dry stage. Among Dipteran, Calliphoridae was the most abundant insect family collected on monkey
carcasses while Sepsidae was the most abundant on rat and rabbit carcasses. The number of insect
taxa collected was lower on rat carcasses compared to that of rabbit and monkey carcasses which
were obvious in the first week of decomposition. Analysis on the successional pattern of insect revealed
that some taxa have a clear visitation period while the others especially Coleopteran did not show a
clear succesional pattern and many gaps existed along the colonization period. For future succession
study, replication of animal and repetition of experiment is crucial to produce a clearer insect
successional pattern.
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PP50
Acarology Unit, Infectious Diseases Research Centre, Institute for Medical Research, Jalan Pahang,
50588 Kuala Lumpur, Malaysia
Objective: To determine distribution of arthropods in processed rice products such as rice flour and
rice cereal-based infant food. Methods: Random samples of rice flour and rice cereal-based infant
food purchased from commercial outlets were examined for the presence of arthropods using a modified
Berlese Tullgren Funnel Method. Mites were mounted prior to identification and weevils were directly
identified. Results: For non-expired products, infestation was found in 6.7% of rice flour and none
was found in rice cereal-based infant food samples. The arthropods found in the flour samples were
Cheyletus spp., Suidasia pontifica, Tarsonemus spp., Tyrophagus putrescentiae, Sitophilus
granarius and Sitophilus oryzae. Others which cannot be identified were Oribatid and Prostigmatid
mites. The most common mites in rice flour were Tarsonemus spp. (69.1%), followed by S. pontifica
(18.2%). For expired products, only one sample of rice cereal-based infant food was infested and the
infestation was by mites of the family Tydeidae. Conclusions: This study demonstrates the presence
of 4 allergenic species of S. pontifica, T. putrescentiae, S. granarius and S. oryzae in rice flour.
These arthropods can contribute to the incidence of anaphylaxis upon consumption by atopic individuals.
There was no infestation of arthropods in rice cereal-based infant food surveyed except for an expired
product in a moderate rusty tin container.
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PP51
It is not uncommon to culture only non-sporulating molds from clinical specimens. In the absence of
spores, these molds could not be identified by cellular morphology and diagnostic labs report these
molds as non-sporulating molds to the clinicians. Clinical significance of these molds is unknown. In
the current work, 20 non-sporulating molds grew from 8 dermatological specimens (skin, hair and nails)
and 12 systemic specimens (blood cultures and other body fluids) were identified using polymerase-
chain-reaction (PCR) methods. DNA extraction was performed using commercial DNA extraction
kits. The internal transcribed spacer region of the ribosomal DNA was amplified using ITS1-ITS4
primers. The purified PCR products were sequenced and the sequences were analysed with BLAST
program. Using this method, non-sporulating molds from nail specimens were identified as Phoma
species and Fomitopsis pinicola while Xylaria feejeensis, Phoma species and Dicyma olivacea
were molds isolated from skin specimens. Non-sporulating molds associated with diseased nails were
identified as Microsporum canis and Phoma species. A dermatiacous fungi, Bipolaris species and
Aspergillus niger were identified from a PD fluid and CSF specimens, respectively. Various
environmental fungi grew in blood cultures and were identified as Pseudozyma parantartica,
Schizophylum commune, Xylaria feejeensis, Fomitopsis ostreiformis, Eutypella species, Penicillium
species and Coprinellus species. It can be concluded that the non-sporulating molds are mostly
clinically insignificant saprophytic fungi. However, established pathogen such as M. canis, emerging
fungal pathogen such as S. commune and opportunistic pathogenic fungi such as P. parantartica and
Phoma species were also among the non-sporulating molds.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 117
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PP52
Shamilah Hisam1, Gan Chee Chean1, Nor Parina Ismail1, Sim Wei Wei2, Chua Hock Hin3,
Chai Ming Cheng2 and Phui Nyat Fung4
1Parasitology Unit, Infectious Diseases Research Centre, Institute for Medical Research, 50588 Jalan Pahang,
Kuala Lumpur
2Dept O&G, Sarawak General Hospital, Malaysia
3Dept Infectious Disease, Sarawak General Hospital, Malaysia
4Parasitology Laboratory, Dept Pathology, Sarawak General Hospital, Malaysia
The patient in our case report was a 27 year old female who presented with complaints of excessive
menstrual bleeding attributed to uterine fibromyoma. Her blood picture showed a microcytic
hypochromic anemia and an ultrasound of the liver showed normal liver echogenicity with smooth
outline with no significant abnormality. Elective laparotomy myomectomy was made and she was
subjected to a full blood work-up prior to the surgery including a stool microscopic examination. Some
ova showed the characteristic ova morphology of Fasciola hepatica or Fasciolopsis buski. However,
a definite diagnosis cannot be obtained solely on egg morphology and an inconclusive clinical history.
Further test with ELISA fascioliasis was negative. This patient was treated with praziquantel, 40 mg/
kg body weight. Subsequent examination of fecal samples showed absence of eggs. Epidemiology of
foodborne trematode infection is greatly dependent upon a suitable ecology for the intermediate snail
host to breed, and close proximity between fresh water and human habitation. Water pollution,
deforestation, climate change and aquaculture may be factors for declining prevalence. This is a rare
case report that warrants further epidemiological survey of the village of the infected person. Perhaps
trematode infections in Malaysia are not truly rare because some villages are still devoid of major
changes in their flora and fauna.
118 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
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PP53
Prolonged state of oxidative stress (caused by overproduction of free radicals than antioxidants) has
been one of the contributory factors of cancer. Free radicals that are generated by hosts immune
cells to kill the invading parasites implicate parasitic infections as a possible cause of oxidative stress.
The activity of free radicals namely reactive oxygen species (ROS) can cause secretion of metabolites
in serum such as malondialdehyde (MDA, metabolite of lipid peroxidation), hydrogen peroxide (H2O2,
indicates hydroxyl radical level) and AOPP (advanced oxidation protein product). Previously, we have
reported that colorectal cancer patients (majority with B. hominis infection) exhibited high level of
AOPP. This indicates that the possibility of intestinal parasitic infection in facilitating cancer growth
via oxidative protein damage should not be ruled out. However such studies have not been reported
in breast cancer. This study compares MDA, H2O2 and AOPP in subjects infected with intestinal
parasites alone and breast cancer patients with and without intestinal parasitic infection. All intestinal
parasite infected subjects and breast cancer patients showed high level of oxidative stress compared
to the healthy individuals. The levels of H2O2 in breast cancer patients with infection were significantly
higher compared to patients without infection. This implicates that intestinal parasitic infections in cancer
are generally detrimental regardless of the cancer types. To date this is the first study to report on
the effect of intestinal parasitic infection towards oxidative damage in breast cancer. Thus, cancer
patients who are mainly known for immunodeficiency should be subjected for intestinal parasitic
screening.
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PP54
Zaridah MZ1, Nor Azah MA1, Abd Majid J1, Mohd Faridz Z1, Nik Yasmin NY1 and Rohani A2
1Medicinal Plants Division, Forest Research Institute Malaysia, Kepong, 52109 Selangor Darul Ehsan
2Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur
Larvicidal bioassay of the essential oil of Ocimum tenuiflorum was carried out with two mosquito
vectors, i.e., Aedes aegypti and Culex quinquefasciatus as the test organisms. Results indicated
that the late third instar larvae of Cx. quinquefasciatus (LC50 31.34 ppm) were more susceptible to
the essential oil when compared to Ae. aegypti (LC50 44.32 ppm). The essential oil of Ocimum
tenuiflorum obtained by hydrodistillation of the fresh aerial parts was analysed by a combination of
capillary GC and GC-MS. The most abundant component of the essential oil was methyl eugenol
(68.7%). Other compounds that were present in appreciable amounts were -caryophyllene (11.6%),
-cubebene (5.6%), methyl isoeugenol (3.8%), eugenol (2.1%) and endo-borneol (1.7%). We suggest
that the existence of these compounds may contribute in the larvicidal activities shown.
PP55
Zaridah MZ1, Nor Azah MA1, Abd Majid J1, Mohd Faridz Z1, Nik Yasmin NY1 and Rohani A2
1Medicinal Plants Division, Forest Research Institute Malaysia, Kepong, 52109 Selangor Darul Ehsan
2Medical Entomology Unit, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur
Citrus hystrix and Citrus aurantifolia of Rutaceae Family were collected from Teluk Intan, Perak
and Sabak Bernam, Selangor, respectively. Both plant species then were hydrodistilled to obtain their
essential oils. The larvicidal bioassay was according to World Health Organization (WHO) guidelines
with slight modification. Late third instar larvae of Aedes aegypti and Culex quenquefasciatus were
chose as the test organisms and the results were observed after 24-h period to determine their median
lethal concentrations (LC50). The result showed that Citrus hystrix essential oil gave median lethal
concentration (LC50) value of 35.90 ppm and 77.67 ppm, against Ae. aegypti and Cx quinquefasciatus,
respectively. C. aurantifolia essential oil gave median lethal concentration (LC50) value of 17.67 ppm
and 12.11 ppm against Ae. aegypti and Cx quinquefasciatus, respectively. From the results, it showed
that C. aurantifolia demonstrated higher larvicidal activity than C. hystrix for both mosquito species.
The essential oil of both plant species were analysed by a combination of capillary GC and GC-MS
leading to the identification of their major chemical compounds.
120 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
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PP56
Microsatellites are short tandem repeats (STRs) that are well known as useful molecular markers for
genome mapping studies due to their high abundance in the genome. STR densities have been shown
to vary among different taxa and genomic regions. In this study, we compared genome-wide distribution
of STRs in the protozoan parasites Eimeria maxima and E. tenella. Sequences for both of these
genomes, which were estimated to be 42Mb and 52Mb in size respectively, were retrieved from publicly
accessed databases. Tandem Repeats Finder (TRF) was used to identify all possible STRs in these
two genomes, which were then further processed by Tandem Repeats Analysis Program (TRAP). E.
maxima was found to have a higher average relative density of STRs with 102398.59 compared to E.
tenella with 95343.43. The distribution of different repeat type classes showed that both genomes
are rich in trinucleotide repeats consisting ~51% of the total STR density in E. maxima and ~52% in
E. tenella. From all of the trinucleotide repeat types, AGC is found to be the most abundant in both
genomes where it represents ~94% and ~99% of the total trinucleotide in E. maxima and E. tenella,
respectively. The second highest density of repeat class in E. maxima is tetranucleotide with the most
abundant repeat type AAAT (~65%), while heptanucleotide has the second highest density of repeat
class in E. tenella with AAACCCT as the most abundant repeat type. These highly abundant repeat
patterns may indicate their functional and evolutionary importance in these two genomes.
~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~ 121
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PP57
Izazy-Nur MJ1,2, Mat-Isa MN1, Suzuki Y3, Blake DP4, Watanabe J5 and Wan KL1,2
1Malaysia Genome Institute, UKM-MTDC Technology Centre, 43600 UKM Bangi, Selangor DE
2School of Biosciences and Biotechnology, Faculty of Science and Technology,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor DE
3Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo,
Tokyo, Japan.
Chicken coccidiosis is a serious disease that causes huge economic losses to the poultry industry
worldwide. Eimeria maxima is recognised as one of the seven Eimeria spp., obligate intracellular
parasites from the Apicomplexa phylum, which are responsible for this disease. The major method of
control of coccidiosis has been chemotherapy. However, due to the widespread of drug resistance,
new targets for intervention are needed. This will require a more in-depth knowledge of the biology
of this parasite. Sequencing of full-length cDNA clones may contribute to this effort. In this study,
approximately 20 million reads were generated from full-length cDNA clones of E. maxima sporozoites
using the Solexa sequencing technology. The assembly of these reads was performed using the Velvet
program. Different hash lengths were tested and the hash length of 51 was found to give the most
optimum assembly resulting in 3404 contigs. Blastx analysis showed that 33.23% of the contigs had
significant similarity (E-value < 10-5) to sequences available in the GenBank non-redundant database.
Most of the significant hits (85.27%) matched with sequences from apicomplexan parasites especially
Toxoplasma gondii and other Eimeria spp., implying that these sequences are conserved and play
important roles in the phylum. Further analysis can be carried out on the remaining contigs with no
significant match as they may represent novel sequences exclusive to E. maxima.
122 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
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PP58
Chandrawathani P1, Chang KW2, Nurulaini R1, Waller PJ4, Adnan M1, Zaini CM1,
Jamnah O1, Khadijah S1,3 and Vincent N2
1Veterinary Research Institute, 59, Jalan Sultan Azlan Shah, 31400, Ipoh, Perak
2Headquarters, Department of Veterinary Services Malaysia, Ministry of Agriculture and Agro-based Industry,
Wisma Tani, Block Podium Lot 4G1, Precinct 4, 62630 Putrajaya
3School of Biological Sciences, University Sains Malaysia, Minden, Penang
4 National Veterinary Institute, Uppsala, Sweden
This study was conducted to evaluate the anthelmintic effect of Neem (Azadirachta indica) on
nematode parasites of sheep. Twelve Santa Ines cross bred sheep from a government farm were
randomly selected and equally divided into control (n = 6) and treated groups (n =6). Faecal egg counts
(FEC) using the modified McMaster technique and the FAMACHA score for assessing clinical anaemia
were carried out daily and recorded for 6 weeks. At the end of the study all the animals were
slaughtered and the total worm count (TWC) was done. The results of FEC showed that there was
no significant difference between the control and treated group (p = 0.081). However, worm burden
estimations showed that the number of parasites was significantly higher in the control group compared
to the treated group (p < 0.05). This result indicated that feeding Neem had an effect on worm numbers
in sheep, but was not reflected in their faecal egg counts. Further work is needed to reconfirm the
effect of Neem on helminth infections of sheep.
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PP59
A variety of plants have been traditionally used by local farmers to treat helminth infection in ruminants.
Previous studies had described a range of in vitro tests that can be conducted to evaluate the
anthelmintic efficacy of any potential plants. More anthelmintic efficacy bioassays should be conducted
on any potential plants especially local species to give more alternatives to the local farmers. In this
study, established larval motility assay was modified based on the principle of egg hatch assay and
larval migration test. Experimental plant, T. catappa was used to determine the anthelmintic potential
against three different targeted species namely H. contortus, T. colubriformis and C. curticei to
prove the effectiveness of this modification. Species differentiation and identification were first
conducted followed by the in vitro study. Larvae was distributed at a concentration of 50 L3 (n=50)
per well in 96-multiwells plates before being incubated with diluted crude aqueous extract of T. catappa
at a ratio of 1:1 at 20C for 3 hours and 5 hours respectively. After 3 hours of incubation, reduction
percentages for T. colubriformis, C. curticei and H. contortus were 70%, 63% and 73% respectively
while at 5 hours of incubation, reduction percentages for each species were 77%, 67% and 80%
respectively. The control showed no reduction in terms of motility and survivality of the larvae with
standard deviation (SD) at 5-10%. These findings shows that with this modification bioassay, the
anthelmintic potential of T. catappa was successfully evaluated and more trials should be conducted
in future.
124 ~~~~~~~~~~~~~~~~~~~~~~~~ Climate Change and its Impact on Public Health ~~~~~~~~~~~~~~~~~~~~~~~~
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PP60
This study focuses on the detection and prevalence of infection of filarial parasites in dogs, cats and
humans in Selangor. Two study areas were chosen as sites for blood sample collection; Kampung
Sungai Bumbun, an Orang Asli village in Pulau Carey and the SPCA Selangor animal shelter in Ampang
Jaya. Microscopic examination of the animal blood samples detected 3 different species of microfilariae
which are; Brugia pahangi, Dirofilaria immitis and Dirofilaria repens. The rate of filarial infection
was determined as 14.3% (1 out of 7) and 28.6% (6 out of 21) in cats and dogs respectively. The
rate of filarial infection was also much lower in the SPCA where only 6.3% (1 out of 16) animals
were infected, compared to Pulau Carey which showed an infection rate of 50% (6 out of 12). The
overall prevalence of these filarial parasite infections in the study areas was 25% (7 out of 28). No
microfilariae were detected upon microscopy of the human blood samples, but testing using a
BRUGIArapid test kit showed a positive result in 1 out of 16 human samples tested (6.25%). PCR
confirmed the species as B. pahangi. From this study, it was demonstrated that the prevalence of
filarial parasites in animals is still high in rural areas such as the Orang Asli village in Pulau Carey.
The finding of B. pahangi infection in a human subject also provides further evidence to prove that
the natural infection of humans with B. pahangi (unreported to date) can occur.
PP61
Noor Aslinda Ummi Awang Besar1, Mohd Nawi Bin Sulaiman1, Junaidi Bin Ibrahim1
and Mahani Yusof2
1Kinta Health District Office, 2Perak Health Department
The adult population and species composition of mosquitoes collected in malarious area in Kinta District
were described. A total of 562 mosquitoes representing 4 genera and 10 species were collected using
human landing catch (Bare Leg Catch), outdoor during 2009 and 2010. Anopheles maculatus was
the most common species (57%) followed by Anopheles barbirostris (23%), Culex sp. and others in
very small percentage. An. maculatus was collected most often in March 2010 and present throughout
the year. Over 2009 and 2010, there are no malaria outbreak in Kinta District. In 2009, 22 malaria
cases were recorded and only 5 localities are from malaria prone areas. Malaria cases were decrease
in 2010, only 15 malaria cases were reported. The identification of vector species and knowledge of
their ecology and behaviour is essential for epidemiologic studies, the design and implementation of
vector control strategies.
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PP62
Manorenjitha Malar S1, Zairi J2, Nor Azlina K3 and Norita AK3
1Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia (USM)
2VectorControl Research Unit, School of Biological Sciences, Universiti Sains Malaysia (USM)
3Animal Research Unit (ARU), Advanced Medical and Dental Institute (AMDI),
Ficus religiosa which is the subject of the present study has been used for centuries by traditional
practitioners in India to treat various diseases. Each part of the tree is believed to posses medicinal
values. Hence, it was decided to investigate the larvicidal activity of aqueous extract of Ficus
religiosas stem bark against laboratory strain Aedes aegypti larvae. Bioassay was conducted under
laboratory condition at the selected doses of 500, 750, 1000, 2500, 5000, 7500, 10000, 12500 and 15000
ppm to determine the LC50 and LC90 value at 24, 48 and 72 hours post treatment. The findings indicate
that high concentration of Ficus religiosa stem bark aqueous extract is needed to suppress 50% of
the larval population of Aedes aegypti. Furthermore, it was also noted that with the exception of
control group, no larvae from the treated groups developed into the pupal stage during the test.
PP63
Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia
A cross-sectional study among rural Malay and Aboriginal children from four primary schools in Lipis
and Raub districts of Pahang state, Malaysia, was carried out to determine the prevalence and potential
risk factors associated with Blastocystis hominis infection. Stool samples were collected from 300
schoolchildren aged 6 12 years (150 males and 150 females) and examined for the presence of B.
hominis by using direct smear observation after in-vitro cultivation in Jones medium. The overall
prevalence of B. hominis was found to be as high as 25.6%. Outputs of the univariate and multivariate
analyses showed that low levels of mothers education (OR=3.29; 95%CI=1.55, 6.98; P<0.01) and
absence of piped water supply (OR=2.83; 95%CI=1.58, 5.04; P<0.001) were the significant risk factors
of B. hominis infection. There was no significant difference in the prevalence of this parasite among
the subjects according to age, sex, race, family size, family income, parental employment status, living
with animals, and presence of gastrointestinal symptoms. In conclusion, B. hominis is prevalent among
rural children and the important factors that determine the infection were the sources of drinking water
and mothers educational level. Interventions of clean water supply and health education especially to
mothers are required.
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ACKNOWLEDGEMENTS
The Organising Committee of the 47th Annual Conference of Malaysian Society of Parasitology and
Tropical Medicine would like to thank the following for their support and contribution:
SPONSORS
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CORPORATE MEMBERS
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LIST OF PARTICIPANTS
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15, Jalan SS5A/11, Kelana Jaya, 47301 Petaling Jaya, Selangor Darul Ehsan, Malaysia
6-03-7877-2611 6-03-7877-2610 [email protected] www.bio-diagnostics.com.my
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Research Reagents
Molecular and cell biology reagents RNAi libraries. Cytokines and growth factors FBS and culture media primary
and secondary antibodies biochemical and ELISA assay kits. Services peptide and gene synthesis custom-made polyclonal and
monoclonal antibodies SNP and genotyping plasmid DNA purification common buffers and solutions subcloning site-directed
mutagenesis GST- and His-tag protein purification.
A-1-6, Pusat Perdagangan Kuchai, No. 2, Jalan 1/127, Off Jalan Kuchai Lama, 58200 Kuala Lumpur
Tel: (03) 7982 0322, Fax: (03) 7987 4566, Email: [email protected], Website: www.i-dna.com.my
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