Shukor Md Nor

Molecular identification of blood meal sources of ticks (Acari, Ixodidae) using cytochrome b gene as a genetic marker

Abstract Blood meal analysis (BMA) from ticks allows for the identification of natural hosts of ticks (Acari: Ixodidae). The aim of this study is to identify the blood meal sources of field collected on-host ticks using PCR analysis. DNA of four genera of ticks was isolated and their cytochrome b (Cyt b) gene was amplified to identify host blood meals. A phylogenetic tree was constructed based on data of Cyt b sequences using Neighbor Joining (NJ) and Maximum Parsimony (MP) analysis using MEGA 5.05 for the clustering of hosts of tick species. Twenty out of 27 samples showed maximum similarity (99%) with GenBank sequences through a Basic Local Alignment Search Tool (BLAST) while 7 samples only showed a similarity range of between 91–98%. The phylogenetic trees showed that the blood meal samples were derived from small rodents (Leopoldamys sabanus, Rattus tiomanicus and Sundamys muelleri), shrews (Tupaia glis) and mammals (Tapirus indicus and Prionailurus bengalensis), supported by 82–88% bootstrap values. In this study, Cyt b gene as a molecular target produced reliable results and was very significant for the effective identification of ticks’ blood meal. The assay can be used as a tool for identifying unknown blood meals of field collected on-host ticks.

Species composition of Amphibians and Reptiles in Krau Wildlife Reserve, Pahang, Peninsular Malaysia

A revised and updated checklist of the herpetofauna of the Krau Wildlife Reserve (KWR), Pahang is presented, which includes 61 species of amphibians and 65 species of reptiles. Five species of amphibians are listed under revised names i.e. Hylarana labialis , H. picturata , Ansonia latiffi , Microhyla mantheyi and Rhacophorus norhayatae ; and two species are listed as new locality records for KWR: Hylarana cf. siberu and Theloderma licin . The amphibians and reptiles recorded in KWR are approximately 56% and 22% of the total number of species recorded from Peninsular Malaysia, respectively.

Molecular Phylogeny of a tick, Ixodes granulatus (Acari: Ixodidae) based on cytochrome oxidase subunit I (COI) marker

Identification of a local species of tick, Ixodes granulatus from the family Ixodidae is essential because it has potential to be vector for spotted fever group (SFG) rickettsia and tick thypus. The aim of this study is to portray the relationships among several populations of I. granulatus collected from different species of animal hosts and localities in Peninsular Malaysia. Polymerase Chain Reaction was conducted by amplifying mitochondrial DNA marker, namely cytochrome oxidase subunit I (COI) sequences from 15 individual ticks that attached to five different hosts caught from three different localities. Confirmation of the species identity was accomplished using BLAST program. Neighbor-joining (NJ) and Maximum Parsimony (MP) tree based on COI sequences were constructed by using PAUP 4.0b10 to identify the relationship among species. The result of this study showed a high genetic heterogeneity between I. granulatus and other species of the same genus (7.2-23.7%). Furthermore, a low intraspecific variat...

Prevalence of on-host ticks (Acari: Ixodidae) in small mammals collected from forest near to human vicinity in Selangor, Malaysia

Abstract Ticks are important vectors that transmit a variety of pathogenic microorganisms known to be medically important worldwide. Many vertebrate groups have become host to this organism, and their presence and abundance are an indicator of the condition of both host and its habitat. This study was conducted to determine ticks infestation and its prevalence on small mammals residing in the recreational forests (RF) and semi-urban (SU) residential areas which have encountered Leptospirosis outbreak and cases in Hulu Langat, Selangor Malaysia. Trapping of the small mammals involved deploying two hundred cage traps in a systematic one-hectare plot (100 m × 100 m), as well as along the stream and forest trails at random. Ticks were extracted from the captured individual hosts. Identification of the tick species was performed based on morphological features and molecular approach using 16S rDNA and COI (cytochrome oxidase subunit I) genes. A total of 278 individuals of small mammals belonging to 15 species (13 Rodentia, 1 Scandentia and 1 Insectivora) were captured in the study areas. From these, 34 individuals from eight small mammal species were infested with ticks. The most infested host species was Mullers giant Sunda rat (Sundamys muelleri) with 5.80% (n=16). Ticks prevalence was slightly higher in RF with 6.40% (n=18) compared to SU with 5.80% (n=16). A total of 107 adult ticks (103 female and 4 male) were collected from the infested host. Ixodes granulatus was the most dominant tick species encountered (70.40%, n=85), followed by Dermacentor sp. (18.60%, n=20), while Amblyomma sp. was the least abundant (2%, n=2). This study provides information on tick species present and tick burden on small mammal hosts within the study areas. Our findings suggest that the visitors to the recreational forests and the residents of the semi-urban area were not only exposed to Leptospirosis bacteria but also tick bites and potentially tick-borne disease, therefore, precaution should be taken to avoid contact with small mammal hosts.

Molecular identification of hard ticks (Ixodes sp.) infesting rodents in Selangor, Malaysia

This study aims to identify hard ticks (Ixodes sp.) infesting rodents in three different sites in Selangor, Malaysia using a molecular approach. A total of 11 individual ticks infesting four different host species (Rattus tiomanicus, Rattus ratus, Maxomys surifer and Sundamys muelleri) were examined based on its morphological features, followed by molecular identification using mitochondrial 16S rDNA gene. Confirmation of the species identity was accomplished by using BLAST program. Clustering analysis based on 16S rDNA sequences was carried out by constructing Neighbour-joining (NJ) and Maximum parsimony (MP) tree using MEGA 7 to clarify the genetic identity of Ixodes sp. Based on morphological features, all individual ticks were only able to be identified up to genus level as most of the samples were fully engorged, damaged and lacked morphological characters. However, molecular analysis of samples revealed 99% similarity with Ixodes granulatus from the GenBank database. Thus, the result of this study showed that all these ticks (Ixodes granulatus) were genetically affiliated to a monophyletic group with highly homogenous sequences.This study aims to identify hard ticks (Ixodes sp.) infesting rodents in three different sites in Selangor, Malaysia using a molecular approach. A total of 11 individual ticks infesting four different host species (Rattus tiomanicus, Rattus ratus, Maxomys surifer and Sundamys muelleri) were examined based on its morphological features, followed by molecular identification using mitochondrial 16S rDNA gene. Confirmation of the species identity was accomplished by using BLAST program. Clustering analysis based on 16S rDNA sequences was carried out by constructing Neighbour-joining (NJ) and Maximum parsimony (MP) tree using MEGA 7 to clarify the genetic identity of Ixodes sp. Based on morphological features, all individual ticks were only able to be identified up to genus level as most of the samples were fully engorged, damaged and lacked morphological characters. However, molecular analysis of samples revealed 99% similarity with Ixodes granulatus from the GenBank database. Thus, the result of this study s...

Diet of tropical insectivorous birds in Lowland Malaysian rainforest

ABSTRACT Despite the importance of diet in avian ecology, knowledge of bird diet in tropical regions, particularly South-East Asia, is limited. Assessing predator–prey relationships of how energy flows across ecosystems provides insight into niche segregation of sympatric species. We examined induced-regurgitation samples from 15 species of insectivorous birds – 12 babblers and three flycatcher-like species – from the Krau Wildlife Reserve, central Peninsular Malaysia. The main objective of this study was to compare and characterise the diet of similar trophically insectivorous birds so as to examine the occurrence of dietary overlap or segregation. The majority of identified prey items belong to the following arthropod taxa: Coleoptera (53%), Hymenoptera (19%), Blattodea (11%), and Araneae (11%). The diet of all bird species slightly overlapped (p = 0.070) but network analysis yielded a relatively low value (Ro = 0.53), indicating dietary divergence may have occurred among the studied bird species, possibly reflected by the variation in proportion of arthropod groups. A high proportion of Coleoptera and three other arthropod groups (i.e. Hymenoptera, Blattodea and Araneae) in the diet of studied birds could be related to their richness and abundance in aerial leaf-litter, the substrate preferred by most babblers. Dietary differences may further be explained by variations in bill shapes and sizes, likely corresponding to prey size. Unique foraging strategies (utilising different vertical strata, microhabitats, and attack manoeuvres) could also lead to dietary separation among sympatric species. Flycatcher-like species were expected to flycatch different prey items (families or lower taxa) versus the gleaning birds (most babblers). This study successfully discerned more details on the diets of insectivorous birds inhabiting Malaysian forests, providing a foundation for future studies on the ecology and biology of understorey birds.

Niche shift in three foraging insectivorous birds in lowland Malaysian forest patches

With the rapid growth of agricultural areas globally, forest birds increasingly encounter fragmented landscapes in which forest patches are surrounded by an agricultural plantation matrix, yet how birds respond behaviourally to this fragmentation is poorly understood. Information on microhabitat requirements of birds is scarce, but nevertheless essential to predicting adaptation of bird species to the patchy landscapes. We investigated foraging patterns of three tropical insectivorous birds, Green Iora Aegithina viridissima, Pin-striped Tit-Babbler Macronus gularis and Chestnut-winged Babbler Cyanoderma erythropterum, to determine whether they vary in foraging methods in different forest patches. Our study area encompassed old-logged lowland forest; one continuous forest and three forest patches. Observations were performed for 15 days every month for a period of 13 months. Information on foraging height, substrate, attack manoeuvres, and foliage density was collected independently for each foraging bird individual. All three species used different foraging substrates and attack manoeuvres in different habitat types. The Green Iora frequently used lower strata when foraging in forest patches as opposed to continuous forest, while the Pin-striped Tit-Babbler tended to forage in more dense vegetation in patches. Only Chestnut-winged Babbler displayed complete foraging plasticity across all study parameters. Different habitat features (e.g., edges, microclimates) between continuous forest and forest patches significantly influenced the foraging strategies of the study species. These changes in foraging strategies suggest that some Malaysian forest birds (e.g. generalist species) can respond behaviourally to fragmentation and habitat loss. Although continuous forest has critically important characteristics that need to be conserved, remnant forest patches are also important as ecological movement corridors and foraging grounds for birds.

Molecular characterization of pathogenic Leptospira sp. in small mammals captured from the human leptospirosis suspected areas of Selangor state, Malaysia

Leptospirosis is caused by the spirochetal bacterium Leptospira of which rodents are considered the most important reservoir. This study aims to determine and characterize virulent Leptospira species among rodents and small mammals found in human settlements and recreational spots within the Hulu Langat and Gombak districts of Selangor, Malaysia; regions that frequently report probable human leptospirosis cases. Molecular analysis revealed an overall Leptospira detection rate of 14.3% among the 266 small mammals captured, and the human settlements were found to have the highest number of isolates (15.1%), followed by recreational sites (14.5%). The molecular characterization conducted based on the lipL32, secY genes and MLST revealed that the strains belonged to four different species, including; Leptospira interrogans (29; 76.3%; ST50, ST238, ST243), L. kirschneri (5; 13.15%; ST110), L. borgpetersenii (3; 8%; ST143) and L. weilii (1; 2.63%; ST242). The study revealed genotypes of circulating strains among small mammals in Malaysia, which include Leptospira locus ST110 L. kirschneri, ST 50 L. interrogans, ST143 L. borgpetersenii and ST242 L. weilii. Among the small mammals studied, 17/105 (16.2%) Rattus norvegicus, 7/59 (11.9%) of Rattus rattus, 5/24 (20.8%) of Maxomys whiteheadi, 4/18 (22.2%) of Sundamys muelleri, 2/22 (9%), Tupaia gliss, 2/16 (12.5%) Rattus tiomanicus and 1/4 (25%) of Suncus murinus carried pathogenic leptospires. The data from the present study may imply that, in addition to rodents, other small mammals also serve as maintenance hosts for Leptospira. Hence, much remains unknown about Leptospira maintenance hosts, and there is need for further investigation to ascertain the prevailing serovars of pathogenic Leptospira in Malaysia. This will assist in the development of efficient diagnostic assays with improved microscopic agglutination test (MAT) panels, and in the implementation of suitable prevention and control measures.

Genetic distance of Malaysian mousedeer based on mitochondrial DNA cytochrome oxidase I (COI) and D-loop region sequences

Mousedeer is one of the primitive mammals that can be found mainly in Southeast-Asia region. There are two species of mousedeer in Malaysia which are Tragulus kanchil and Tragulus napu. Both species can be distinguish by size, coat coloration, and throat pattern but clear diagnosis still cannot be found. The objective of the study is to show the genetic distance relationship between T. kanchil and T. napu and their population based on mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and D-loop region. There are 42 sample of mousedeer were used in this study collected by PERHILITAN from different locality. Another 29 D-loop sequence were retrieved from Genbank for comparative analysis. All sample were amplified using universal primer and species-specific primer for COI and D-loop genes via PCR process. The amplified sequences were analyzed to determine genetic distance of T. kanchil and T. napu. From the analysis, the average genetic distance between T. kanchil and T. napu based on locus COI and D-loop were 0.145 and 0.128 respectively. The genetic distance between populations of T. kanchil based on locus COI was between 0.003-0.013. For locus D-loop, genetic distance analysis showed distance in relationship between west-coast populations to east-coast population of T. kanchil. COI and D-loop mtDNA region provided a clear picture on the relationship within the mousedeer species. Last but not least, conservation effort toward protecting this species can be done by study the molecular genetics and prevent the extinction of this species.

Species composition and richness of amphibians in logged forests at Hulu Terengganu, Peninsular Malaysia

A study was done to compare amphibian species composition and richness between a disturbed forest due to logging/dam construction and a 30-year old logged forest at Hulu Terengganu, Peninsular Malaysia. This study was conducted from August to October 2014, using drift fenced-pitfall traps. The upstream (UP) and downstream riverine area (DP) of a dam called Puah Dam (PD) represented the disturbed forest habitat, while Sg. Deka Wildlife Reserve (SDWR) represented a 30-year old regenerating logged forest. There were six amphibian species found at SDWR, while four species were recorded at UP and DP.