Malaysian Species Research Articles

Rare orchid species in Malaysia: New records, recollections and amended descriptions.

Paphiopedilum exul, Calanthe chrysoglossoides, and Luisia brachystachys are reported here as new records for Malaysia, whereas Bryobium cordiferum subsp. borneense, Habenaria rostellifera, and Taeniophyllum rugulosum are three rare orchid species recollected from Sarawak, Perlis, and Perak, respectively. This paper highlights brief descriptions and photographic illustrations of each species for easy identification. Besides, notes on morphological comparisons with the closely related species and artificial taxonomic keys are included as well.

The Compilation Records of Fireflies (Coleoptera: Lampyridae) Diversity and Distribution and Display Trees Throughout Malaysia

The populations of fireflies (Coleoptera: Lampyridae) are increasingly being threatened, and it is, thus, a significant problem in Southeast Asia, particularly Malaysia. Fireflies and their habitat must immediately be protected before they go extinct. Simpson’s Diversity Index used in this paper review to measure the diversity of firefly species across Malaysia. The Simpson’s Diversity Index showed that the richness and evenness of firefly species in Malaysia is infinite diversity with D = 0.2255. The authors compiled and reviewed the studies on the firefly species to incorporate available information/data and emphasised their preferred habitat/display trees in response to the issue. Therefore, this paper was also able to track down records of fireflies’ species’ distribution through the previous studies in eight states in West Malaysia (Peninsula), namely Johor, Kelantan, Kuala Lumpur, Pahang, Perak, Negeri Sembilan, Selangor, and Terengganu, as well as two states in East Malaysia (Borneo), namely Sabah and Sarawak. It indicates that firefly species, especially from the general Pteroptyx (Pteroptyx tener species), are widely distributed in Peninsular Malaysia and East Malaysia. Based on the records from the study conducted by previous researchers, it was found that the population of fireflies is declining in some areas in Malaysia, and so are their habitats and host/display trees that have suffered the same decline. Perhaps, this paper will help broaden human beings’ geographical understanding/knowledge and create awareness which eventually leads to conservation actions of firefly species and their host/display trees.

Morphological Variation between Hatchery Bred and Wild Caught Anabas testudineus in Malaysia.

This study was designed to characterise the cultured and wild populations of Anabas testudineus in Malaysia using morphological parameters. Fish samples from the East and West coast of the country were obtained from fishermen (wild samples) and well-recognised climbing perch farmers in Kedah, Kelantan, Johor and Selangor. The Truss network method was applied to obtain necessary data and analysed to examine phenotypic variation between the cultured and wild stocks. Results obtained suggest that each hatchery population belonged to a distinct stock as revealed by their separate clustering into individual unique groups. However, an extensive overlap was observed in the wild population suggesting similarity of origin. The most important morphological parameters for the discrimination of the two populations are the homologous landmark B (i.e., snout to insertion of the pelvic fin) and C (i.e., above the eye to insertion of the pelvic fin). Genetic characterisation of the A. testudineus is needed to complement the findings of this study and establish a baseline for the development of a selective breeding programme for the fish species in Malaysia.

Multiplex PCR Assay for the Identification of Four Species of the Anopheles Leucosphyrus Sub-Group in Malaysia.

Simple SummaryPlasmodium parasites cause malaria. The bites of infected female Anopheles mosquitoes, known as “malaria vectors,” transmit the parasites to people. To prevent the spread of malaria, precise mosquito species identification is essential. This study aims to develop a quick and accurate method for identifying the Anopheles species (An. introlatus, An. latens, An. cracens, and An. balabacensis), which have been incriminated as vectors for simian malaria in Malaysia. Overall, six primers targeting the internal transcribed spacer 2 (ITS2) region of each species were designed for this assay. This study is helpful for the researchers or vector-related field workers to correctly identify the mosquitoes for control activities.The Leucosphyrus Group of mosquitoes are the major simian malaria vectors in Malaysia. Accurate species identification is required to help in curbing the spread of simian malaria. The aim of the study is to provide an accurate molecular method for identifying the four important Anopheles vector species found in Malaysia. Mosquito specimens were collected from various localities in Malaysia, where simian malaria cases were reported. DNA from 122 mosquito specimens was tested to develop a multiplex polymerase chain reaction (PCR) assay. The specificity of this assay was tested against other mosquito species. Molecular identification of the species was further confirmed by analysing the internal transcribed spacer 2 (ITS2) DNA region of the specimens. Anopheles balabacensis and An. latens showed two distinct clades in the phylogenetic tree. The multiplex PCR assay was developed based on the ITS2 region for the identification of Anopheles introlatus (298–299 bp), Anopheles latens (197–198 bp), Anopheles cracens (421–426 bp), and Anopheles balabacensis (224–228 bp). This method will be useful to accurately identify the major Anopheles Leucosphyrus Group species in Malaysia, which are difficult to identify morphologically, to determine the correct vector as well as its geographical distribution.

Blood meal analysis of Anopheles vectors of simian malaria based on laboratory and field studies

Blood feeding and host-seeking behaviors of a mosquito play an imperative role in determining its vectorial capacity in transmitting pathogens. Unfortunately, limited information is available regarding blood feeding behavior of Anopheles species in Malaysia. Collection of resting Anopheles mosquitoes for blood meal analysis poses a great challenge especially for forest dwelling mosquitoes. Therefore, a laboratory-based study was conducted to evaluate the potential use of mosquitoes caught using human landing catch (HLC) for blood meal analysis, and subsequently to document blood feeding behavior of local Anopheles mosquitoes in Peninsular Malaysia. The laboratory-based experiment from this study revealed that mosquitoes caught using HLC had the potential to be used for blood meal analysis. Besides HLC, mosquitoes were also collected using manual aspirator and Mosquito Magnet. Overall, 47.4% of 321 field-caught Anopheles mosquitoes belonging to six species were positive for vertebrate host DNA in their blood meal. The most frequent blood meal source was human (45.9%) followed by wild boar (27.4%), dog (15.3%) and monkey (7.5%). Interestingly, only Anopheles cracens and Anopheles introlatus (Leucosphyrus Group) fed on monkey. This study further confirmed that members of the Leucosphyrus Group are the predominant vectors for knowlesi malaria transmission in Peninsular Malaysia mainly due to their simio-anthropophagic feeding behavior.

The genera Coccus and Prococcus (Hemiptera: Coccomorpha: Coccidae) in China, with two new combinations and descriptions of two new species.

The Chinese soft scale species in the genus Coccus Linnaeus, 1758 (Hemiptera: Coccomorpha: Coccidae) were studied. Coccus cambodiensis Takahashi and the Malaysian species C. cameronensis Takahashi are considered to be non-congeneric with Coccus hesperidum Linnaeus, 1758, the type species of Coccus, and are transferred to Prococcus Avasthi, 1993, as Prococcus cambodiensis (Takahashi, 1942), comb. n. and Prococcus cameronensis (Takahashi, 1952), comb. n. The generic diagnosis of Prococcus is revised. In the genus Coccus, two new species are described and illustrated based on adult females: Coccus nanningensis Cao Feng, sp. n. from Guangxi, China, on Ficus carica (Moraceae), and Coccus cephalotaxus Cao Feng, sp. n. from Shannxi, China, on Cephalotaxus sinensis (Taxaceae). Identification keys to separate adult females of Prococcus from Coccus, the 14 species of Coccus found in China, and all three species of Prococcus are provided.

In silico analysis of Gracilaria changii proteins for potential bioactive peptides

Gracilaria changii is a red seaweed species in Malaysia with high protein content (12.57% (dry basis)). Thus, G. changii proteins are potential precursors for producing bioactive peptides. To date, no study has been reported on the potential of G. changii proteins as potential precursors for bioactive peptides. In this study, fourteen G. changii proteins were selected as potential precursors of bioactive peptides using in silico approach. It was found that the most potential bioactivity was dipeptidyl peptidase-IV (DPP IV) inhibitory and angiotensin-I converting enzyme (ACE) inhibitory activities. Papain, ficin and stem bromelain were used for in-silico proteolysis. Stem bromelain was found to be more effective in terms of the release of fragments with a given activity. Furthermore, two tripeptides (ACF and YCL) were screened as novel and promising bioactive peptides. The characteristics of both peptides were also analyzed using PeptideRanker, PepCalc, Peptide Cutter, ToxinPred, AllerTop and AHTpin bioinformatic tools. The bioinformatic tools predicted that both peptides were non-toxic, non-allergen and highly potential. The present work suggests that G. changii can serve as a potential source of bioactive peptides and these findings can provide a basis for future in-vitro and in-vivo study of bioactive peptides from G. changii proteins.

Distribution of seahorse species in Malaysia, harvesting trends and conservation concerns – A review

Seahorse is a unique fish belonging to the genus Hippocampus. It has a horse-like head, tubular mouth, and the body is covered with rings of armoured plates until the tip of the prehensile tail. Seahorses are traded for human consumption and ornamental purposes. However, the dry product of seahorses has a major demand for traditional Chinese medicine (TCM). The ornamental products such as keychains, earrings and brooches also support a growing ‘curio’ trade. There is a limited amount of published data on seahorse distribution in Malaysia despite increasing pressure of exploitation, this study was carried out to review the available information, identify knowledge gaps and draw attention towards sustainability of the seahorse populations. To date, twelve seahorse species have been reported in Malaysia. Most of these species are found in East Malaysia. In Sabah, 11 out of 12 tropical seahorse species have been encountered. This does not include the Japanese dwarf seahorse, Hippocampus mohnikei. Among the eleven species of seahorses found in Sabah are seven species of common size seahorses, H. barbouri, H. comes, H. histrix, H. kelloggi, H. kuda, H. spinosissimus and H. trimaculatus and four species of pygmy seahorses, H. bargibanti, H. denise, H. satomiae and H. pontohi, that were previously identified as H. severnsi. This study highlights the importance of conservation of wild population of seahorses and the need for farming as a means of lessening the exploitation of wild stocks while meeting the human demand.

Legislative Approach in Managing Environment Pollution by Alien Fish Species in Malaysia

River pollution has been a major problem in Malaysia and significantly affects the environment. One of the contributing issues would be the spread of invasive alien fish species that has given negative impacts on the environment and ecological disaster, the human health, harming the native animals and plants, as well the economy. Based on the National Committee on Invasive Alien Species Report in 2018, that 46 invasive alien species from a total of 130 species in the established main invasive alien species lists in Malaysia. The alien fishes have been known to make their presence in our dams, lakes and rivers but not much attention has been paid to their existence. Based on these facts, it is proven that the issue poses a significant risk hence an urgent attention is required in managing this pollution through proactive legal measures to enforce strictly the existing law so as to prevent the halt of the spread of invasive species. A legal doctrinal and non-doctrinal modes of research are used to examine and review on the existing laws and/or policy governing this issue. This research will help to provide reference and strategic planning via legal approach for better controlling the invasion in our Malaysian waters.

Morphological and molecular identification of medically important questing Dermacentor species collected from some recreational areas of Peninsular Malaysia.

Questing is a situation when a tick is seeking to get closer or ambush its potential host. However, information on questing tick species in Malaysia is still lacking, thus the association with tick-borne diseases (TBD) is not completely understood. The aim of this study was to investigate the tick species from five most frequently visited recreational areas in Pahang and Terengganu states, which were recorded to have high potential of TBD cases. By implementing handpick method, a total of 18 males and 15 females belonging to five Dermacentor Koch, 1844 species, were collected, namely D. compactus Neumann 1901, D. tricuspis (Schulze, 1933), D. auratus Supino 1897, D. steini (Schulze, 1933), and D. falsosteini Apanaskevich, Apanaskevich & Nooma respectively. The specimens were collected and identified based on morphological characters prior to obtaining the molecular data of COI and 16S rDNA. The D. compactus was the most abundant species collected in this study, while D. falsosteini was the least. All species were distinctly separated on the Neighbor Joining and Maximum Parsimony tree topologies and supported with high bootstrap values. Furthermore, a low intraspecific variation (0.00 - 0.01) was observed amongst the individuals of the same species in both genes. Meanwhile, each Dermacentor species was genetically different, with interspecific values ranging from 0.13-0.19 and 0.11-0.20 for COI and 16S rDNA. These findings had successfully recorded the tick species that were potentially associated with TBD, and which might be circulated among humans and animals. This study also has some implications on the diversity and geographical extension of Dermacentor ticks, thus should warrant further investigation as a potential vector of tick-borne diseases and public health importance.

Predicting aquatic invasions in a megadiverse region: Maximum‐entropy‐based modelling of six alien fish species in Malaysia

Abstract There has been a recent increase in the occurrence and proliferation of alien fish species in rivers, lakes, and reservoirs of Malaysia, posing a significant risk to native biodiversity. This increase has been largely caused by the unmanaged aquaculture and aquarium pet trade. At present, very little information is available on the diversity and distribution of alien fish species in freshwater ecosystems of Malaysia, as well as their current and future threats. Using the MaxEnt model, the potential habitat suitability and future invasion ranges of six alien fish species (Cichla ocellaris, Clarias gariepinus, Oreochromis mossambicus, Oreochromis niloticus, Poecilia reticulata, and Pterygoplichthys pardalis) in Peninsular and East Malaysia (Borneo) were determined. All six alien fish species investigated, have high potential to invade eight states in Peninsular Malaysia (Johor, Kelantan, Malacca, Negeri Sembilan, Pahang, Perak, Selangor, and Terengganu), and much of East Malaysia. The results also indicate that alien fish are likely to invade the majority of the 444 protected areas in the country, particularly those located in Sabah and Sarawak, which are known for their high fish diversity and endemism. Minimum temperature of the coldest month (Bio 6) and isothermality (Bio 3) significantly influence potential invasion habitats for all six alien fish species in Malaysia. The country's hot and humid climate makes it ideal for the survival and proliferation of alien species except for P. pardalis. These findings will help conservation planners and managers in identifying areas of high concern for alien freshwater fish species in Malaysia, and to pre‐emptively mitigate and manage their future invasion.

Effect of Sugar-Pectin-Citric Acid Pre-Commercialization Formulation on the Physicochemical, Sensory, and Shelf-Life Properties of Musa cavendish Banana Jam

Cavendish banana is the second most cultivated species in Malaysia due to its high potassium and vitamin B6 content. However, there has been little commercialisation of Cavendish banana in jam making to date because of its unappealing physical characteristics and unsavoury taste. In the present study, response surface methodology was used to determine the optimum quantity of sugar (175-275 g)-pectin (3-7 g)-citric acid (150-234 mL) (SPC) on water activity, °Brix, colour analysis, pH, total titratable acidity (TTA), and sensory attributes of formulated banana jams. The amount of sugar-pectin-citric acid was found to have an effect on the TTA and pH of the banana jams. Conversely, water activity and °Brix were only affected by the amount of sugar. The lightness, L* of the banana jams was influenced by the quantity of pectin and the volume of citric acid added. Sensory analysis using 30 panellists showed that there were changes in the colour, taste, aroma, texture, and overall acceptability of banana jam depending on the amount of sugar used. In a shelf life study, banana jams stored at 4 °C were found to have a longer shelf life compared to those stored at 25 °C. Overall, the optimal formulation for a high-quality banana jam was 281.79 g of sugar, 4.13 g of pectin, and 264.66 mL of citric acid. This study constitutes the first report on the potential pre-commercialisation formulation for Cavendish banana jam production.

Mitochondrial Barcodes of Three Malaysian Butterflies Originating from Taman Negara Endau Rompin Johor, Malaysia

Butterflies are projected as reliable and economical biodiversity indicator. Traditionally, taxonomists identified and classified butterfly species based on highly similar and ambiguous morphological appearances which can result in problematic species identification process leading to misidentification of species. DNA barcoding has been developed for taxonomic identification of butterflies to species level. Nonetheless, this approach is hampered by the paucity of reference barcodes encompassing butterflies of all families and species in Malaysia. This study reported four novel DNA barcodes (two cytochrome oxidase subunit I (COI) and two cytochrome b, cytb) of Malaysian butterflies generated using Sanger sequencing. One barcode (DIB032 COI) supported the species identified by taxonomists whereas the other three barcodes (DIB034 COI, DIB034 cytb and DIB046/049 cytb) deduced that the butterfly species were either misidentified or unidentified up to species level due to the lack of reference barcodes in GenBank. The four novel DNA barcodes were deposited in GenBank under the accession number MT210226.1 (DIB032 COI), MT210227.1 (DIB034 COI), MT210228.1 (DIB034 cytb) and MT210229.1 (DIB046/049 cytb).

Introduction of Invasive Peacock Bass (Cichla Spp.), Its Rapid Distribution and Future Impact on Freshwater Ecosystem in Malaysia

Abstract Malaysia is recognised among the mega-diversity countries with the abundance of various freshwater species. Malaysian freshwater biodiversity is under threat after the introduction of an invasive peacock bass. The presence of this predator could challenge the coexistence of the native species. In the current review, the most important aspects regarding peacock bass threats to the biodiversity future of local species are addressed. Four non-native species of peacock bass are presently recognised in Malaysia. To date, the peacock bass species expand to all parts of Peninsular Malaysia with no records yet found in Kelantan and Borneo. Invasion success is more susceptible in Malaysian lakes (84.38%) compared to the dams, rivers and reservoirs. The mode of rapid expansion of this species is highly connected to the sport fishing and anglers. A propagule pressure, favourable habitat, prey abundance and feeding behaviours are responsible factors for the successful establishment of this species in Malaysia. The species are generalist feeders and piscivorous in nature. It is difficult to eradicate peacock bass since it provides physical jobs (recreation and tourism). However, the damage the species will cause in future is unpredictable. At this point, proper management of this species must be implemented to reduce its population. This could be achieved through updating checklists of freshwater bodies, improving monitoring systems and public awareness.

Taxonomy, Evolutionary and Dispersal Events of Pig-Tailed Macaque, Macaca nemestrina (Linnaeus, 1766) in Southeast Asia with Description of a New Subspecies, Macaca nemestrina perakensis in Malaysia.

The pig-tailed macaque, Macaca nemestrina, which is distributed in Peninsular Malaysia, Thailand, Borneo, and Sumatra, has been the subject of unstable and changing taxonomic entity in the M. nemestrina group. This species is involved with a human-macaque conflict in Malaysia and at the same time played an important role in the ethnozoological culture of Malaysian. Even so, comprehensive phylogenetic, population genetics and biogeographical analysis of M. nemestrina in Malaysia are non-existent after decades of intensive research on the genus itself. Thus, we conducted the first comprehensive genetic study of M. nemestrina in Malaysia, based on three mitochondrial loci-Cytochrome b (567 bp), D-loop (398 bp), and COI (577 bp)-from 27 individuals representing Malaysia, plus an additional 26 sequences of Southeast Asian macaques from Genbank. Comparative biogeographical analysis in this study supports the positions of M. nemestrina in M. nemestrina groups as opposed to the silenus or Sulawesi groups. Results from this study also indicate that Bornean populations are the first extant lineages to separate from the other examined lineages of M. nemestrina, M. leonina, M. pagensis, and M. siberu in Southeast Asia. Molecular clock analysis suggested that M. nemestrina arrived in the Malay Peninsula about 0.32 million years ago (MYA). Our results indicate that the population of pig-tailed macaque from Perak (west Peninsular Malaysia) differs genetically based on all phylogenetic and population genetic analyses. Morphologically, Perak's pig-tailed macaque shows brighter coloration than M. n. nemestrina. Thus, we proposed a new subspecies for Perak's pig-tailed macaque as Macaca nemestrina perakensis distributed in the state of Perak, Peninsular Malaysia. This research helps resolve the taxonomic position and population genetics of pig-tailed macaque in Malaysia, which contribute directly to conservation and management of the species in Malaysia.

Public knowledge and perceptions of the impacts and importance of alien fish species in Malaysia: implications for freshwater biodiversity and conservation

Public knowledge and perceptions of the impacts and importance of alien fish species in Malaysia: implications for freshwater biodiversity and conservation

Phytoremediation of Vegetable Leachate by Nymphaea Nouchali

Leachate normally contains high concentration of pollutants and regulated under stringent discharge standards. Conventional treatments of leachate are no longer adequate to ensure compliance with the standard. Phytoremediation is a relatively new approach to treat leachate contaminated by organic and inorganic substances. Nymphaea Nouchali is an abundant and easy-growing free-floating aquatic plants found in the rice fields, reservoir and polluted ponds. Nymphaea and Nuphar are the most commonly found species in Malaysia. In the last few decades there has been an increasing interest in the study of phytoremediator plants which can serve as an alternative for leachate treatment. The main objective of this study is to determine the effectiveness of organic removal from vegetable leachate (VL) using Nymphaea Nouchali. The results exhibited that after 7 days of phytoremediation, sample R0.5 shows excellent removal of biological oxygen demand (BOD), chemical oxygen demand (COD), colour and suspended solid (SS) to meet the allowable limit.

Rapid in-situ detection kit (RisK): Development of loop-mediated isothermal amplification (LAMP) assay for the rapid identification of selected invasive alien fish in Malaysian freshwaters

Invasive alien fish species have become a silent treat towards the ecosystem especially the native fish population in Malaysia. There has been a need to develop rapid identification methods that can aid management teams in identifying fish species that are not native to our ecosystem. Current visual identification methods are highly tedious and require time, delaying action towards curbing the invasion. The LAMP assay successfully identified six popular invasive fish species in Malaysia. None of the LAMP assays showed false positives and the Limit of Detection of the LAMP primers were highly sensitive and could detect DNA samples up to 1 × 10−15 ng/μl. The LAMP primers designed were highly specific to the target species and did not amplify non target species. DNA sequencing was done to ensure the accuracy of LAMP assay results. This study demonstrates that LAMP is a suitable tool in species identification efforts of invasive fish species in Malaysia.

MINERAL CONTENT AND PHYTOCHEMICAL PROPERTIES OF SELECTED CAULERPA SPECIES FROM MALAYSIA

Seaweeds are abundant and are an important renewable resource for the coastal community since they are rich sources of minerals and natural bioactive compounds. Among them, the Caulerpa species under green seaweeds is widely consumed by locals in Southeast Asia. Edible seaweeds are often associated with various health benefits. However, in Malaysia, limited studies have been done on the mineral and antioxidant levels of seaweeds, especially for the Caulerpa species. Thus, the present study aimed to evaluate the variations of minerals and antioxidants of selected Caulerpa species in Malaysia. The samples were collected in two study sites, which were Blue Lagoon, Port Dickson and Merambong Shoal, Johor. Two species, C. racemosa and C. manorensis were recorded from Merambong Shoal, Johor, while five species were recorded namely C. sertularioides, C. racemosa var lamourouxii, C. lentillifera, C. racemosa var cylindracea and C. racemosa at Blue Lagoon, Port Dickson. Among all the Caulerpa species from both study sites, C. manorensis possessed the highest phosphorus content (139.54 mg/100g) while C. racemosa from both Merambong and Blue Lagoon Port Dickson’s calcium (Ca) content was significantly highest at 2318.33 – 2406.66 mg/100g. All the Caulerpa species possessed high sodium (Na) content ranging 9338.30 – 21748.30 mg/100g. For micronutrients, the highest zinc content was recorded in C. manorensis (3.40 mg/100g) while the copper content was significantly highest in C. racemosa from Port Dickson (1.05 mg/100g). The methanol extract of C. sertularioides possessed phytochemical attributes of high antioxidant activity (DPPH EC50 = 24.16 mg/mL) with a high FRAP value being recorded in C. lentillifera (27.09 mg TE/100g). The TPC and TFC were highest in C. lentillifera with 57.95 mg GAE/100g and 1506.41 mg QE/100g respectively. The present study revealed that Caulerpa species contained constituents with significant mineral compositions and phytochemical attributes suitable for pharmaceutical and nutraceutical uses.  

Development of species-specific SCAR markers for identification and authentication of three rare Peninsular Malaysian endemic Coelogyne (Orchidaceae) orchids

Background: Coelogyne kaliana, Coelogyne stenochila and Coelogyne tiomanensis are three valuable rare orchid species endemic to Peninsular Malaysia, currently rampantly traded illegally via the internet and through local nurseries, which label them as hybrids to avoid enforcement detection. Drastic measures to ensure the continued existence of their populations in the wild should be introduced as they are rapidly diminishing into extinction, including the development of rapid and accurate species-specific identification tools. These three orchid species are highly similar morphologically and currently it is impossible to distinguish among them without their reproductive structures. Methods: RAPD-based species-specific SCAR markers were developed to distinguish and authenticate the identity of these three endemic Peninsular Malaysian Coelogyne species. Results: Three SCAR markers were successfully developed in this study. SCAR marker primer pair, CKL_f / CKL_r was specific to C. kaliana as it produced a unique single band of 271 bp but not in C. stenochila and C. tiomanensis. SCAR marker primer pair CST_f / CST_r amplified a single band of 854 bp in C. stenochila and two bands of different sizes (372 bp and 858 bp) in C. tiomanensis, but no amplification in C. kaliana. The third SCAR marker primer pair, CTI_f / CTI_r produced a single band (about 500 bp) for both C. stenochila and C. tiomanensis, but showed no amplification in C. kaliana. Conclusions: Although not all these SCAR markers were species amplification specific, they could be used to discriminate among the three Coelogyne species effectively. Accurate species identification is one of the most important steps to allow a proper management plan to be established in the effort to conserve these three endangered orchid species of Peninsular Malaysia. Besides, it could effectively put a stop to the illegal trading of these rare endangered orchid species worldwide.