Genetic diversity of the S-type small subunit ribosomal RNA gene of Plasmodium knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia

Objective:To determine the genetic diversity of Plasmodium(P.)knowlesi isolates from Sabah,Malaysian Borneo and Peninsular Malaysia,targeting the S-type SSU rRNA gene and including aspects of natural selection and haplotype.Methods:Thirty-nine blood samples infected with P.knowlesi were collected in Sabah,Malaysian Borneo and Peninsular Malaysia.The S-type SSU rRNA gene was amplified using polymerase chain reaction,cloned into a vector,and sequenced.The natural selection and haplotype of the S-type SSU rRNA gene sequences were determined using DnaSP v6 and illustrated using NETWORK v10.This study's 39 S-type SSU rRNA sequences and eight sequences from the Genbank database were subjected to phylogenetic analysis using MEGA 11.Results:Overall,the phylogenetic analysis showed no evidence of a geographical cluster of P.knowlesi isolates from different areas in Malaysia based on the S-type SSU rRNA gene sequences.The S-type SSU rRNA gene sequences were relatively conserved and with a purifying effect.Haplotype sharing of the S-type SSU rRNA gene was observed between the P.knowlesi isolates in Sabah,Malaysian Borneo,but not between Sabah,Malaysian Borneo and Peninsular Malaysia.Conclusions:This study suggests that the S-type SSU rRNA gene of P.knowlesi isolates in Sabah,Malaysian Borneo,and Peninsular Malaysia has fewer polymorphic sites,representing the conservation of the gene.These features make the S-type SSU rRNA gene suitable for comparative studies,such as determining the evolutionary relationships and common ancestry among P.knowlesi species.

Heavy metal pollution assessment in marine sediments in the Northwest coast of Sabah,Malaysia

Heavy metal contents along the Northwest coast of Sabah were determined to interpret the pollution level in the marine sediment. The metal abundance is regulated by the physico-chemical properties such as the average sediment pH(7.82, 9.00 and 8.99), organic matter(0.62%, 1.60%, and 2.27%), moisture content(25.00%, 29.70%, and 15.00%) and sandy texture in Kota Belud, Kudat and Mantanani Island,respectively. The major elements show Ca>Fe>Mg>Al>Mn for all study sites, while the heavy metals show Ni>Cr>Zn>Cu>Co>Pb, Cr>Ni>Zn>Cu>Pb>Co and Zn>Pb>Cr>Ni, for Kota Belud, Kudat and Mantanani Island, respectively. The pollution degree of heavy metals was evaluated by using the Sediment Quality Assessment(SQA). The SQA parameters indicated none to moderate pollution in Kota Belud that shows Class 0, Class 1 and Class 2 pollution. The parameters also indicated none to low pollution in Kudat and Mantanani Island that show only Class 0 pollution. The enrichment factor(EF) suggested minor to moderately severe metal enrichment by anthropogenic sources in Kota Belud, whereas only minor enrichment in Kudat and Mantanani Island. The modified pollution degree(MCD<1.5) and pollution load index(0 PLI<1) indicating only low pollution level in the marine sediments for all study sites. The objectives of this study are:(1) to determine the physico-chemical parameters of sediments,(2) interpret the heavy metal contents and(3) evaluate the sediment quality.

Comparison of Fatty Acid and Proximate Composition between <i>Holothuria edulis</i>and <i>Holothuria scabra</i>Collected from Coastal Water of Sabah, Malaysia

The nutritional values of different species of sea cucumber are greatly concerned because of their dietary and curative properties. In this study, two species of sea cucumber, Holothuria edulis, a low-valued noncommercial species, and Holothuria scabra, a high-valued commercial species were selected to compare its proximate composition and fatty acids. H. edulis a prevalent species in coastal water of Sabah is not commercially importance like H. scabra. Sea cucumbers were captured live from the Sabah marine habitat. All samples were immediately eviscerated, freeze-dried and stored at 4°C in until analyzed. Silylating agent N, O-Bis(trimethylsilyl) trifluoroacetamide (BSTFA) was used to derivitization of fatty acid prior to gas chromatography-mass spectrometry (GC-MS) analysis. Proximate compositions (%), such as moisture, crude protein, crude lipids and ash were carried using standard methods. Major fatty acids in H. edulis and H. scabra were saturated fatty acid (SFA) accounted for 83.95% and 98.60%, respectively and dominated with Palmitic acids. Polyunsaturated fatty acid (PUFA), arachidonic acid of 16.05% was found only in H. edulis, but absent H. scabra. Proximate compositions (dry weight) were varied greatly within these two species. Moisture, crude protein, crude lipids and ash of 85.5%, 70.5%, 1.37% and 1.27% respectively were obtained in H. edulis. On the other hand 84.5% of moisture, 51.2% of crude protein, 0.27% of crude lipids and 4.44% of ashes were determined in H. scabra. Significantly higher protein (p H. edulis compare to H. scabra could be the choice of option for the utilization of this non commercial species as nutraceutical industry and also alternatives to reduce the pressure on heavily exploited species of H. scabra.

Chemical composition of the Tertiary black shales of West Sabah, East Malaysia

The X-ray fluorescence and ICP methods were used to analyze 60 outcrop samples of black shale, of which 15 were collected from Belait, 15 from the Setap Shale, 15 from Temburong, and 15 from the Trusmadi for- mations. The average compositions of the shales from the study area are 64.62%, 63.95%, 62.32%, 63.84% SiO2, 1.84%, 2.14%, 2.04%, 1.99% MgO, 2.55%, 3.12%, 2.89%, 2.72% K2O, 0.32%, 0.30%, 0.32%, 0.53% CaO, 5.86%, 6.06%, 7.14%, 6.60% Fe2O3, 207×10-6, 180×10-6, 213×10-6, 200×10-6 Rb, and 56×10-6, 49×10-6, 50×10-6, 32×10-6 Sr for the Setap Shale, Temburong, Belait and Trusmadi samples, respectively. The high Rb/Sr ratios of 3.8, 3.7, 4.2, and 6.1 are attributed to the lowest contents of Sr due to reducing conditions prevailing. The high Rb/K ratio sug- gests either brackish marine or rapid deposition that prevented equilibrium between Rb and K in the shales and ma- rine waters.

Occurrence and Distribution of Marsh Clam, <em>Polymesoda</em>spp. in Marudu Bay, Sabah, Malaysia

Marudu Bay is part of the Tun Mustapha Marine Park, the largest marine protected area within the Malaysian region of the Coral Triangle Initiative. The bay is known for its diversed fisheries resources including bivalves. Although some of these bivalve species are commercially important, their occurrence, distribution and stock status in the bay are not well documented. Hence, the current study was conducted to determine the occurrence, distribution and the stock status of marsh clam, Polymerasoda spp. in the mangrove swamp situated at the southernmost of the Marudu Bay. Samplings were carried out at the mangrove swamps which covered an area of 500 sequare meter per sampling site. Two marsh clams species, Polymesoda erosa and P. expansa were found to inhabit the sampling sites. In general, juvenile marsh clams were noticed to dominate the seaward mangrove swamp, whereas the high tidal regions were dominated by adults. The current study also suggested a link in the distribution and the morphometric measurements of the marsh clams with the sediment grain size.

Spatial Distribution of Dissolved Inorganic Nutrients and Phytoplankton around Kota Kinabalu Wetland, Sabah, Malaysia

Kota Kinabalu Wetland is importantly habited of mangrove, diverse aquatic flora and fauna as well as feeding stop for migratory birds. This wetland is inundated with the tidal flow, as connected with a small river and nearby coastal areas, thus. A study was carried out to determine the spatial distribution of dissolved inorganic nutrients and phytoplankton diversity at Kota Kinabalu (KK) Wetlands. Five stations, in which river mouth of Likas Bay, river channel (two stations) and inundated area (two stations) in KK Wetland were selected for this study. In-situ parameters of water, water for nutrients and phytoplankton samples were collected from May 2019 until October 2019. The highest concentration of nitrate (0.115 mg/L) was recorded at inundated area of wetland (S5) while the lowest nitrate concentration (0.0047 mg/L) was found at river (S3) flowing towards wetland. The concentrations of ammonia (0.2004 to 2.311 mg/L) were recorded relatively higher at every station compared to other dissolved inorganic nutrients (DIN). The concentration of phosphate was determined in the ranges of 0.0089 - 0.0513 mg/L. Nitrate, ammonia and phosphate showed no significant difference (P = 0.737) in terms of DIN concentration at all five sampling stations during the study period. Twenty-four genera of phytoplankton were identified, dominated by diatoms (55.29%), followed by dinoflagellates (24.95%), Chrysophyta (11.15%), Spirotrichea (5.28%) and Cyanophyta (3.33%). Dominating species throughout the study period include Chaetoceros sp., Pseduo-nitzschia sp., and Cylindrotheca closterium, Peridinium quinquecorne and Alexandrium sp. Phytoplankton species compositions were observed the highest in river mouth area in July with the highest density of 12.115 × 104 cells/mL. The study showed that nutrient concentration was insignificant (P = 0.614) in altering the phytoplankton density, as influences with the tidal water.

Variations in the Physicochemical Water Parameters and Phytoplankton Community in Coastal Water of Kudat, Sabah, Malaysia

This study was conducted to observe the abundance and distribution of phy-toplankton and temporal and spatial variation physico-chemical water parameters in coastal water of Kudat, Sabah, Malaysia. Water samples and in-situ water quality parameters were taken from five selected locations from May 2019 to February 2020. The sampling location was selected based on human-induced activities such as Marina Resort’s Jetty (ST1), Sabah Ports’ Jetty (ST2), aquaculture cage/pent (ST3), river’s mouth (ST4) and Landung Ayang’s water village (ST5). Water parameters: pH, salinity (ppt), dissolved oxygen (mg/L) temperature (°C) and depth (m) were recorded once every month from the selected station. Identification of phytoplanktonic species and cell density (cell/mL) were determined from collected water samples. Significant differences (p < 0.05) between physicochemical parameters to months were observed during the study period. However, in spatial variations, significant differences (p < 0.05) of pH, dissolved oxygen (mg/L) and temperature (°C) observed. A total of 21 phytoplankton species were identified from the study area, where 4 species belonged to Dinophyceae (HABs blooming species) and 17 species belonged to Bacillariophyceae. The influences of physicochemical water parameters were not significant in phytoplankton diversity and abundance. In addition to these parameters, the nutrients in the water might have important roles in the blooming of phytoplankton, which are essential and vital to address in this type of research.

Groundwater Resources of a Small Tropical Island,Sabah,Malaysia:Numerical and Simulation Analysis

Groundwater occurs as a thin lens of freshwater floating over seawater in small islands aquifers.The impacts of climate change and pressure on island resources have been the greatest threats faced by the small islands.This paper represents a case study of groundwater response towards the climate change and pressure on island resources in Manukan Island of Sabah,Malaysia.A modeling software SEAW-

Assessment of Spatial and Temporal Variations of Water Quality for Future Mariculture Operation in Ambong Bay, Sabah, Malaysia

Study was conducted with the aim to understand the temporal and spatial variations of water quality parameters (temperature, salinity, pH, DO, TSS, NO3-, NO2-, NH3-N and PO4-P, and phytoplankton cell density) in Ambong Bay, Sabah, Malaysia in order to provide reference for future mariculture development in the bay. Samplings were carried out once a month in two stations (coastal and open sea) within the bay for 12 months period from September 2015 to August 2016. Results showed that there were significant differences in pH and NO2- when compared spatially, whereas salinity, DO, TSS, phytoplankton cell density, NO3-, NH3-N, and PO4-P were temporally significant. The fermentation processes by anaerobic bacteria, organic acids from decaying vegetation and acidic clays in the mangrove soils might explain the significant spatial differences in pH and NO2-. The bay was dominated by dinoflagellate, Prorocentrum spp. (mean abundance of 16.23% and 24.44%, respectively) a potentially toxic algae species. Correlation matrix showed that NH3-N was positively correlated with PO4-P (r = 0.475, p < 0.05) but negatively correlated with salinity (r = –0.517, p < 0.01). Besides, salinity was positively correlated with DO (r = 0.505, p < 0.05) and TSS (r = 0.408, p < 0.05). In addition, DO and TSS were also positively correlated (r = 0.451, p < 0.05). Phytoplankton cell density was positively correlated with TSS (r = 0.644, p < 0.01). In general, the water quality in Ambong Bay is within the standard values permitted by the Malaysia Marine Water Quality standard for marine life, fisheries, coral reefs, recreational and mariculture (Class 2), except for NO3-. In conclusion, any mariculture operation to take place in Ambong Bay in the near future should take the temporal variation of the water quality into account. Moreover, effects of toxic phytoplankton to culture fishes should also be taken care and monitored frequently.

Distribution Pattern of Dissolved Inorganic Nutrients and Phytoplankton Diversity in River Estuary, Kota Kinabalu Sabah, Malaysia

Nutrients are influenced by anthropogenic activities and consequently change the diversity and density of phytoplankton. The spatial distribution of dissolved inorganic nutrients and phytoplankton diversity and density were determined in river estuary, Kota Kinabalu, Sabah, Malaysia. Samples of water and phytoplankton were collected from six locations of river estuary during the month of May 2019 to October 2019. The highest abundances of 86% diatoms were determined during the study period. This was followed by 13% and 1% of dinoflagellates and Cyanobacteria respectively. The most dominance species of Skeletonema costatum, with density of 1186.69 × 103 cells/L and Thalassiosira sp. (938.01 ×103 cells/L) were recorded from Station 1 (upstream) and Station 5 (mangrove area) respectively. The Chaetoceros sp. (84.25 × 103 cells/L) was the dominated in Station 6 (control site). Station 4 (residential area) accounted the highest diversity of species such as Skeletonema costatum, Thalassiosira sp., Peridinium sp., Gonyaulax sp., and Cylindrotheca sp. with total cell density of 142.98 × 103 cells/L. The high diversity index of 5.2 to 7.09 and evenness index of 1.81 to 2.87 were determined from this river estuary. Very poor relationship was observed with the cell density and dissolved inorganic nutrients, phosphate (R2 = 0.2437), ammonium (R2 = 0.301) and nitrate (R2 = 0.406). The diversity and abundance of phytoplankton in this river estuary not only depend on nutrients, but might be associated with other environmental factors, tidal fluctuations and rate of discharge of nutrient inputs from surrounding areas. This study suggests that long term monitoring not only the nutrients, but discharges and flushing of nutrient during rainfall and tidal fluctuation together with environmental factors should be considered in order to conclude status on the diversity and abundance of phytoplankton in river estuary.

Spatial and Temporal Variations of Dissolved Inorganic Nutrients and Relationship with Phytoplankton Density in Coastal Water of Kudat, Sabah, Malaysia

Coastal areas of Kudat are dominated by fisheries activities. Development of infrastructure for fishing facilities like jetty, landing centre and other human activities can increase the loading of nutrients in coastal area. High load of nutrients accelerates the blooming of phytoplankton and in long run creates eutrophication. This study was conducted to determine the temporal and spatial variations of nutrients and relationship of phytoplankton density in coastal water of Kudat. Five stations were selected with three replicated in the study area. Phytoplankton samples, water samples and in situ environmental parameters were collected from May 2019 to February 2020 (10 months). The highest concentration of nitrate (NO3) was observed in May 2019, but the highest concentration of phosphate (PO4) observed in July 2019. On the other hand, higher concentrations of phosphate (PO4) were determined than the concentration of nitrate (NO3) in all stations during study period. The distribution of nutrients is due to monsoonal runoff from surrounding areas as well due to anthropogenic activities. Among the 21 species three of them are from harmful algal species and but dominated by diatoms. Poor relationship observed among the dissolved inorganic nutrients and phytoplankton density, indicates that the relationship does not depend on only nutrients but with favourable environmental parameters. Anthropogenic activity can lead to excessive load of nutrients in Kudat coastal water and in long run cause eutrophication problem in ecosystem with potentially larger economic impacts in a long run period.

Indigenous Use and Trade of Dugong (Dugong dugon) in Sabah,Malaysia

引言 儒艮（Dugongidae：Dugong dugon）如果尚未绝迹。其残留种群在一些濒临灭绝的区域有片断性分布。但是，1999年报道马来西亚半岛柔佛州（Johors）附近海域发现一批儒艮死亡的事件之后，儒艮被认为在马来西亚海域仍有存在。该事件使儒艮在马来西亚的生存现状受到关注。重新激起了相关学者对儒艮进行研究的兴趣。在马来西亚东部，儒艮以小片断种群分布于沙巴州的山打根（Sandakan District）、直落文莱（Tcluk Brunei）和古达（Kudat District）沿岸海草丛生的水域。

Health Risk Assessment Due to Heavy Metals Exposure via Consumption of Bivalves Harvested from Marudu Bay, Malaysia

Concern over health risk from consumption of bivalves originating from Marudu Bay is escalating due to the rapid agricultural development surrounding the bay. This has motivated us to estimate the health risk index (HRI) of heavy metals from four commercially important and highly exploited bivalve species which are abundant in the bay. Samples (n = 30) of green mussel (Perna viridis), Asiatic hard clam (Meretrix meretrix), Pacific oyster (Crassosstrea gigas) and marsh clam (Polymesoda expans) were acquired from fishermen in Kg. Teritipan, Marudu Bay. These bivalves were analyzed for heavy metals content using the Inductively Couples Plasma-Optical Emission Spectroscopy (ICP-OES). The study found that the mean contents of Arsenic (As) and Manganese (Mn) in all analyzed bivalves exceeded the permissible limits as well as copper (Cu), lead (Pb) and zinc (Zn) in Pacific oyster, and Zn in marsh clam. It was also noticed that consumption of different bivalve species may bring about health risk from different metals as indicated by varied Total Hazard Index (THI) values. Consumption of the four bivalves was noticed to promote high health risk from As intoxication. Although metal pollution index (MPI) analysis revealed that the bivalves from the bay are currently not seriously impacted by heavy metal pollution, vigorous efforts should be taken to preserve the natural condition of the bay for years to come. There are several ways to minimize health issues from bivalve consumption which include keeping the bivalve natural habitat away from heavy metals pollution by strictly enforcing environmental laws and policies, establishing zones for bivalve fisheries, monitoring heavy metals concentration in bivalve on a regular basis and making depuration process a compulsory requirement in every seafood restaurant throughout the country.

Gas chromatography-mass spectrometry analysis of various organic extracts of Merremia borneensis from Sabah

Objective:To analyse the chemical composition of different extracts of Merremia borneensis(M. borneensis) by gas chromatography-mass spectrometry(GC-MS).Methods:The dried leaves powder was extracted with methanol at room temperature by using Soxhlet extractor.Methanol crude extracts of M.borneensis were extrastel with hexane,chloroform,ethyl acetate and butanol. Results:Qualitative analyses of various organic crude extracts showed that majority of these are flavonoids,terpeniods,alkaloids and glycosides.Most of the identified compounds by GCMS are biologically important.Further the M.borneensii leaf possesses certain characteristics that can be ascribed to cultivation on a domestic plantation.Conclusions:The suitable extracts for respective compounds can be chosen on the basis of above GC-MS analysis.All the major compounds from different extracts are biologically active molecules.Thus the identification of a good number of compounds from various extracts M.borneensis might have some ecological significance.

A Preliminary Assessment of Load Consumption and Solar Power Potential at Kota Belud,Sabah

This paper presents a detailed preliminary assessment of load consumption and solar power potential at the Eco-Tourism Centre of Liogu Ku Silou-Silou(EPLISSI),Kota Belud,Sabah.This initial investigation assessed the feasibility of an off-grid solar PV system at EPLISSI with a suitable solar panel system for project installation and commissioning purposes.Due to the absence of an electrical grid and power supply,no pre-existing electrical appliances could be found in EPLISSI.Hence,an excel-based software,the ESCoBox,was used to produce the load profiles.The input data for this software came from a list of required electrical appliances(LED lights,fans,and phone chargers)and the historical frequency of visitors to EPLISSI.Meanwhile,to assess the solar power potential at EPLISSI,an online simulator known as Global Solar Atlas version 2.3 or GSA 2.3 was used.As an input for the GSA 2.3,the initial solar panel system capacity was set for 0.5 kWp,and then an increment of 0.1 kWp was entered until specific criteria were met.The selection of the suitable size is made when the system can satisfy the daily total average load demand and a specific load fulfillment demand.As a result,it was found that the site requires a total average demand and a total peak demand of 4.60 and 11.87 kWh/day,respectively.From the GSA 2.3 generated report,an off-grid solar PV system with the capacity of 2.50 kWp solar PV can satisfy the daily total average load demand of this area,where the average PV energy output is within the range of between 7.74–9.80 kWh/day or an average of 8.72 kWh/day.In conclusion,this preliminary assessment indicates that installing an off-grid solar PV system in this area is possible.

Sauna Technique, Drying Kinetic Modeling and Effectiveness on Solar Drying Compared with Direct Drying in Drying Process of <i>Kappaphycus striatum</i>in Selakan Island Malaysia

A sauna drying technique—the solar drier was designed and imposed, constructed and tested for drying of seaweed. The seaweed moisture content was decreased around 50% in 2-day sauna. Kinetic curves of drying of seaweed were known to be used in this system. The non-linear regression procedure was used to fit three different drying models. The models were compared with experimental data of red seaweed being dried on the daily average of air temperature about 40℃. The fit quality of the models was evaluated using the coefficient of determination (R2), Mean Bias Error (MBE) and Root Mean Square Error (RMSE). The highest values of R2 (0.99027), the lowest MBE (0.00044) and RMSE (0.03039) indicated that the Page model was the best mathematical model to describe the drying behavior of sauna dried seaweed. The percentage of the saved time using this technique was calculated at 57.9% on the average solar radiation of about 500 W/m2 and air flow rate of 0.056 kg/s.

Temporal and Spatial Distribution of Nutrients and HABs at Coastal Water of Kota Belud, Sabah

A study was carried out to determine the temporal and spatial distribution of nutrients and Harmful Algal Blooming species (HABs) to establish the relationship between nutrient concentrations and HABs density in the coastal water of Kota Belud, Sabah, Malaysia. Eight stations were set up in four transects (each transect had two stations;one 500 meters seawards from shoreline, and another five kilometers seawards from shoreline) along the Kota Belud coastal area. Phytoplankton samples, water samples and in situ environmental parameters were collected during dry and wet seasons. Altogether nine HABs species were identified from the study area, where six species (Pyrodinium bahamense, Prorocentum micans, Neoceratium furca, Prorocentum sigmoides, Dinophysis caudate, and Neoceratium fursus) belonged to Dinophyceae and three species (Thalassionema nitzchoioides, Chatoceros affinis, Rhizosolenia sp.) belonged to Bacillariophyceae. Among nine species, Chatoceros affinis was the most abundance composed of 80.6% of total species recorded from all stations during study period. Among the toxic producing Dinoflagellate, Pyrodinium bahamense bloom (>103 cells/l) was observed during study period. Trong linear relationship (r2 = 0.80) was observed between the cell density and concentration of nitrate nitrogen. Relationship between cell density and phosphate phosphorus was poor (r2 = 0.51). The study showed that increasing in nutrient concentrations resulted in the increasing of HABs density. Nitrate was seen to be more important than phosphate in Kota Belud water as limiting factor of the growth of HABs.

Malaysia＇s Contemporary Political and Economic Relations with Iran

Traditionally, Malaysia＇s relations with the Middle Eastern World boil down to narrow-limited interaction with only a handful of countries, mainly Egypt, Saudi Arabia （before known as Hejaz）, and Turkey. The underlying reasons behind are largely owing to the established ancient relations and religious fulfillments like performing pilgrimage as well as pursuing religious studies in highly respected institutions. Malaysia＇s relation with the Islamic Republic of Iran, however, has shown steady progress of development since the end of 1960s and onwards. Although Iran has become an Islamic Republic after the eruption of the Revolution in 1979, the relations remain intact in spite of the potential expansion of Iran＇s religious ideology, Shi＇ah, among Malaysian Muslim communities. On top of that, with Iran now facing economic sanctions due to its nuclear programs, both countries continue to bolster their relations, especially in political and economic interactions. Contemporarily, both Malaysia and Iran are closely working together to further enhance the level of their relations. The leaders of both countries have quiet regularly exchanged visits and have also actively ventured into promising business opportunities. Iran ranked as the Malaysia＇s third West Asian trading partners along with the United Arab Emirates （UAE） and Saudi Arabia, provides a lot of opportunities for Malaysia to seize onto. In other words, Malaysia＇s contemporary relations with Iran will evolve beyond the dogma of the Iranian Revolution in 1979.

Genetic variations of Chrysomya megacephala populations in Malaysia (Diptera: Calliphoridae)

Samples of Chrysomya megacephala were collected from five sites in Malaysia, namely Penang, Selangor, Pahang, Johor, and Sabah. Molecular analysis was conducted using the cytochrome oxidase gene and ISSR-PCR to identify polymorphic markers for distinguishing the populations and to estimate the genetic relatedness among these populations. Two SNPs were detected at sequence position 952 (M1) and 1120 (M2), involving transitional substitutions with cytosine (C) being replaced with thymine (T) and guanine (G) replaced with adenine (A), but there was no amino acid variation. Penang and Selangor shared the same nucleotide T at the M1 position, while M2 variation was found in Pahang population only. The NJ tree based on the COI sequence showed two main groups with branch A comprised of Penang and Selangor populations while branch B consisted of Johor, Pahang and Sabah populations. ISSR analysis indicated that 18 scorable bands (size 200 to 1800 bp) were amplified with UBC808 and UBC835, 11 bands of which were polymorphic and the rest monomorphic. Sabah population was left out for further analysis due to small number of specimens. UPGMA dendrogram using ISSR data showed two branches, one consisted of Penang and Selangor populations, while the other consisted of Pahang and Johor individuals. The polymorphic loci (P) for Johor Baru were 5.56%, Penang 16.70%, Selangor and Pahang 22.2%. Nei’s H values are respectively 0.018, 0.060, 0.050 and 0.051. Similarly the Shannon index diversity values are respectively 0.028, 0.089, 0.085 and 0.087. The lowest genetic distance (0.0997) was between Penang and Selangor populations, while the highest (0.613) was observed between Penang and Pahang populations. AMOVA shows low genetic differentiation among individuals within populations and that 85.6% of total genetic variance was among individuals between populations. This was also indicated by FST statistic of 0.8562. The gene flow between populations was low with Nm of 0.042, and Mantel’s test indicated a significant correlation between genetic distance and log geographic distance (P < 0.05 from 10,000 randomizations). In conclusion the Penang and Selangor fly populations are similar genetically, compared to the other populations.