To determine the geographic origin of the black fly Simulium suzukii on Okinawa Island, Japan, macrogenomic profiles derived from its polytene chromosomes were compared with those of mainland and other insular populations of S. suzukii and of the isomorphic Simulium tani species complex. The Okinawan population is a chromosomally unique cytoform, designated 'D,' which is essentially monomorphic and differs by about 27 fixed rearrangements from the chromosomal standard sequence for the subgenus Simulium and by two fixed differences from its nearest known relative, representing the type of S. suzukii, on the main islands of Japan. Chromosomal band sequences revealed two additional, sympatric cytoforms of S. suzukii, designated 'A' and 'B,' each with species status, in Korea, and a third cytoform, designated 'C,' on Hokkaido, Japan. A new cytoform, 'K,' of S. tani from Malaysia, representing the type of S. tani, is more closely related to cytoforms in Thailand, as are populations from Taiwan previously treated as S. suzukii but more closely aligned with S. tani and newly recognized as cytoform 'L' of the latter nominal species. Rooting of chromosomal band sequences by outgroup comparisons allowed directionality of chromosomal rearrangements to be established, enabling phylogenetic inference of cytoforms. Of 41 macrogenomic rearrangements discovered in the five new cytoforms, four provide evidence for a stepwise origin of the Okinawan population from populations characteristic of the main islands of Japan. The macrogenomic approach applied to black flies on Okinawa Island illustrates its potential utility in defining source areas for other species of flies including those that might pose medical and veterinary risks.
The widespread nominal black fly Simulium (Simulium) rufibasis Brunetti was reexamined morphologically, chromosomally, and molecularly to determine the status of populations in Japan and Korea with respect to S. rufibasis from the type locality in India and to all other known species in the S. (S.) tuberosum species-group. Morphological comparisons established that the species previously known as S. rufibasis in Japan and Korea is distinct from all other species. Consequently, it was described and illustrated as a new species, Simulium (S.) yamatoense. Simulium yokotense Shiraki, formerly a synonym of S. rufibasis, was morphologically reevaluated and considered a species unplaced to species-group in the subgenus Simulium. Chromosomal analyses of S. yamatoense sp. nov. demonstrated that it is unique among all cytologically known species of the S. tuberosum group and is the sister species of the Taiwanese species tentatively known as S. (S.) arisanum Shiraki. Populations of S. yamatoense sp. nov. included two cytoforms, based on the sex chromosomes. Cytoform A, including topotypical representatives, was found in Kyushu, Japan, whereas cytoform B was found in Korea and Honshu, Japan. Molecular analysis based on the COI mitochondrial gene generally corroborated morphological and chromosomal data that S. yamatoense sp. nov. is a distinct species and, like the chromosomal data, indicate that it is most closely related to S. arisanum, with interspecific genetic distance of 2.92-4.63%.
Ovitrap surveillance was conducted in 2012 and 2006 in Malay and Aboriginal Villages on Carey Island. In each village, standard ovitraps were placed indoors and outdoors at randomly selected houses/locations. All L3 larvae recovered were identified up to species level. Results demonstrated that only larvae of Aedes albopictus were found in all the positive ovitraps placed indoors and outdoors. In 2012, a high ovitrap index (OI) of 66.7% indoor and 84.0% outdoor in the Malay Village; and 62.5% indoor and 88.0% outdoor in Aboriginal Village with an apparent absence of Aedes aegypti. In 2006, a 100% OI was recorded in all ovitraps set indoors and outdoors in both villages.
Our previous study demonstrated that among different habitat sites (mangrove, estuary, river, seagrass and Open Sea) in Johor Strait, Malaysia, seagrass showed highest family diversity and abundance of larval fish. However, it is unclear whether this was due to difference in habitat complexity or water quality parameters.? To test this, larval fish were collected by using a bongo net equipped with a flow meter by subsurface horizontal towing from different habitats in Johor Strait between October 2007 and September 2008.? Various physico-chemical parameters were measured and then examined for any relationship to fish larvae diversity and abundance. Among the 24 families identified from the sites, seven families (Blenniidae, Clupeidae, Mullidae, Nemipteridae, Syngnathidae, Terapontidae and Uranoscopeidae) were significantly correlated with the tested waters quality parameters.? Salinity showed a positive and negative significant correlation with Clupeidae (p < 0.01) and Uranoscopeidae (p < 0.05), respectively. Terapontidae was significantly correlated with dissolved oxygen (p < 0.01), while both Mullidae and Syngnathidae were significantly correlated with pH (p < 0.05). However, a canonical correspondence analysis test indicated weak overall correlation (36.4%) between larval assemblage and in the seagrass-mangrove ecosystem of Johor Strait, Malaysia. This likely indicates that habitat structure was more important in determining larval abundance (highest in the seagrass habitat) as compared to water quality at the tested sites. This study emphasizes the need to conserve seagrass beds as important nursery grounds for various fish larvae to ensure adequate recruitment and ultimately sustainable fisheries management. ?
The final instar larva of Acrogomphus jubilaris Lieftinck, 1964, is described and figured for the first time based on exuviae from four male and one female larvae collected in Sarawak, Malaysian Borneo. The adults of A. jubilaris are very rarely encountered. The larvae, however, are surprisingly common in forest streams in Borneo. It is compared with A. malayanus Laidlaw, 1925 and A. walshae Lieftinck, 1935, and notes on behavior, distribution and habitat are included. A map including all known records of A. jubilaris is provided.
A study on insect succession of monkey carcass in a forested area in Ulu Gombak, Selangor, Malaysia was conducted from 9 May to 18 June 2007. The third instar of the housefly, Musca domestica (Linnaeus) (Diptera: Muscidae) were only found on dry stage of a decomposed (Day-33) monkey carcass (Macaca fascicularis Raffles). This observation revealed that M. domestica maggots were found together with other muscid fly maggots, Hydrotaea (=Ophyra) spinigera (Stein) (Diptera: Muscidae) on dry stage of a carcass. However, the role of M. domestica on forensic entomological study remains unknown. This study recorded the first finding of M. domestica maggots on primate carcass in Malaysia.
The presence of the Afro-Australian genus Conochironomus Freeman, 1961 (Diptera: Chironomidae) in Asia has been recognised only informally. An unpublished thesis included Conochironomus from Singapore, and the genus has been keyed from Malaysia without named species. Here, the Sumatran Conochironomus tobaterdecimus (Kikuchi & Sasa, 1980) comb. n. is recorded from Singapore and Thailand. The species is transferred from Sumatendipes Kikuchi & Sasa, 1980, rendering the latter a junior synonym (syn. n.) of Conochironomus Freeman. Conochironomus nuengthai sp. n. and Conochironomus sawngthai sp. n. are described as new to science, based on adult males from Chiang Mai, Thailand. All species conform to existing generic diagnoses for all life stages, with features from male and female genitalia, pupal cephalic tubercles and posterolateral 'spurs' of tergite VIII providing evidence for species distinction. Some larvae are linked to C. tobaterdecimus through molecular barcoding. Variation in other larvae, which clearly belong to Conochironomus and are common throughout Thailand, means that they cannot be segregated to species. Larval habitats include pools in river beds, urban storage reservoirs, drains with moderately high nutrient loadings, and peat swamps. Endochironomus effusus Dutta, 1994 from north-eastern India may be a congener but may differ in adult morphology, thereby precluding formal new combination until discrepancies can be reconciled. Many problems with vouchering taxonomic and molecular material are identified that need to be rectified in the future.
Human gnathostomiasis is a food-borne zoonosis caused by a tissue nematode of the genus Gnathostoma. The disease is highly endemic in Asia, including Thailand. The freshwater swamp eel (Monopterus albus), the second intermediate host of the gnathostome nematode, has an important role in transmitting the infection in Thailand. Surveys on the infective larvae of Gnathostoma spinigerum based on morphological features in freshwater swamp eels have been performed continuously and reported in Thailand. However, there is still limited molecular data on intra-species variations of the parasite. In this study, a total of 19 third-stage larvae of morphologically identified G. spinigerum were collected from 437 liver samples of freshwater swamp eels purchased from a large wholesale market in Bangkok, Thailand. Molecular characterization based on mitochondrial cytochrome c oxidase subunit I (COI) sequences was performed to elucidate their genetic variations and phylogenetic relationship. Among the 19 infective larvae recovered from these eels, 16 were sequenced successfully. Phylogenetic analyses inferred from the partial COI gene showed the presence of three distinct COI haplotypes. Our findings confirm the presence of G. spinigerum as the main species in Thailand.
Although the microbiome of blood-feeding insects serves an integral role in host physiology, both beneficial and pathogenic, little is known of the microbial community of black flies. An investigation, therefore, was undertaken to identify culturable bacteria from one of Malaysia's most common black flies, Simulium tani Takaoka and Davies, using 16S rDNA sequencing, and then evaluate the isolates for antibiotic resistance and virulence genes. A total of 20 isolates representing 11 bacterial species in four genera were found. Five isolates showed β-hemolysis on Columbia agar, and virulence genes were found in three of these isolates. Some degree of resistance to six of the 12 tested antibiotics was found among the isolates. The baseline data from this study suggest rich opportunities for comparative studies exploring the diversity and roles of the microbiome of S. tani and other Southeast Asian black flies.
We access the molecular diversity of the black fly Simulium nobile De Mejiere, using the universal cytochrome c oxidase subunit I (COI) barcoding gene, across its distributional range in Southeast Asia. Our phylogenetic analyses recovered three well-supported mitochondrial lineages of S. nobile, suggesting the presence of cryptic species. Lineage A is composed of a population from Sabah, East Malaysia (Borneo); lineage B represents the type population from Java, Indonesia; and lineage C includes populations from the mainland of Southeast Asia (Peninsular Malaysia and Thailand). The genetic variation of lineage C on the mainland is greater than that of lineages A and B on the islands of Borneo and Java. Our study highlights the value of a molecular approach in assessing species status of simuliids in geographically distinct regions.
A standardized larval dipping method was used to determine the infestation rates of Culex and other species of mosquitoes in stagnant water at 20 residential areas. This study also examined the associations between Culex distribution and various habitat characteristics across all states in Malaysia. Identification of 7,848 specimens yielded 6 species dominated by Culex quinquefasciatus (82.74%), followed by Cx. vishui (14.39%), Cx. gelidus (2.70%), Lutzia fuscanus (0.11%), Armigeres subalbatus (0.05%), and Anopheles separatus (0.01%). The Culex larvae occurred in stagnant water with pH ranging from 6.4 to 8.2; conductivity, 139.7 to 6635.2 micros/cm; salinity, 0.07 to 3.64 ppt; total dissolved solids, 0.09 to 4.27g/liter; and dissolved oxygen, 5.11 to 8.11 mg/liter. The mean number of Culex larvae was positively correlated with pH, conductivity, salinity, and total dissolved solids. In contrast, the elevation and dissolved oxygen were found negatively correlated with mean number of Culex larvae. This study documented baseline information on the habitat characteristics of Culex species for the 1st time at different residential areas in Malaysia. The findings of this study will be a timely reminder to local authorities that effective control measures should be monitored regularly in order to reduce the nuisance of these mosquitoes and the risks of disease transmission.
Mosquitoes are principal vectors of major vector-borne diseases. They are widely found throughout urban and rural areas in Malaysia. They are responsible for various vector-borne diseases such as dengue, malaria, filariasis and encephalitis. A total of 158 mosquito larvae specimens were collected from the National Zoo, Malaysia, from 11 types of breeding habitats during the study period from end of May 2007 to July 2007. Aedes albopictus was the predominant species (35.4%), followed by Tripteroides aranoides (26.6%), Lutzia halifaxii (11.4%), Aedes alboscutellatus (10.1%), Aedes caecus (8.9%), Armigeres spp. (4.4%), Malaya genurostris (2.5%) and Culex vishnui (0.6%). It is important to have a mosquito free environment in a public place like the zoo. Routine larval surveillance should be implemented for an effective mosquito control program in order to reduce mosquito population.
The megophryid frogs Leptobrachella brevicrus, Leptolalax dringi and Megophrys dringi are species exclusively known from highly localised areas in isolated mountain ranges on Borneo. The tadpoles and adults in this study were collected at the shared type locality for the three species in Gunung Mulu National Park, Sarawak, Malaysia (Borneo). The species identities of larvae were determined via comparison to syntopic adults using DNA barcoding techniques based on partial 16S rRNA mitochondrial gene sequences. The genetic data supported the status of the three taxa as valid species. Descriptions of colouration in life and after preservation, external morphological features, morphometric measurements and ecological notes in comparison to congeneric species are supplied. The tadpoles of L. brevicrus and L. dringi show similar adaptations to a fossorial lifestyle. These include an elongated, vermiform body, a relatively long tail and small eyes. Both were found in the gravel beds of a small mountain stream. In contrast, the larvae of M. dringi are adapted to occupying and feeding at the surface of pools within the stream.
The occurrence of Setaria digitata in a horse is reported for the first time in Malaysia. An 8-year-old Thoroughbred cross mare was referred to the University Veterinary Clinic with the primary complaint of corneal opacity and excessive eye discharge. After initial treatment with Terramycin eye ointment, corneal opacity cleared partially to reveal a moving thread-like cylindrical worm in the anterior chamber of the eye. The parasite was successfully removed surgically, and examination under the light microscope revealed that the isolated worm (length = 45 mm) was a 5th stage larva of S. digitata based on morphological criteria. Confirmation of the species of the worm was through molecular methods. The 12S rRNA gene was PCR-amplified, and the purified amplicon was directly sequenced. Phylogenetic analyses revealed that the isolated roundworm showed 100% sequence similarity with that of S. digitata in NCBI GenBank database (Accession no.: KY284626.1). This report is the first confirmed case of equine ocular setariasis by S. digitata in Malaysia. The current study provides evidence that S. digitata is an etiological agent of ocular infection and its presence in Malaysia.
The composition of fish larvae and their diversity in different habitats are very important for fisheries management. Larval fishes were investigated in a mangrove estuary of Marudu Bay, Sabah, Malaysia from October 2012 to September 2013 at five different sites. Monthly samples of fish larvae were collected at five sampling sites by a plankton net with a mouth opening of 40.5 cm in diameter. In total, 3879 larval fish were caught in the investigated area. The mean density of ichthyoplankton at this area was 118 larvae/100 m(3). The fish larval assemblage comprised of 20 families whereas 13 families occurred at St1, 16 at St2, 16 at St3, 12 at St4 and 16 at St5. The top major families were Sillaginidae, Engraulidae, Mugilidae and Sparidae with Sillaginidae consisted 44% of total larval composition. St3 with 143 larvae/100 m(3) had the highest density amongst the stations which was due to higher abundance of Sillaginidae. Shannon-Wiener diversity index represented significant variation during monsoon and inter-monsoon seasons, peaking in the months December-January and May-June. However, Shannon-Wiener index, evenness and family richness showed significant differences among stations and months (p < 0.05).
It is important to obtain frequent measurements of the abundance, distribution, and seasonality of mosquito vectors to determine the risk of disease transmission. The number of cases of dengue infection has increased in recent years on Penang Island, Malaysia, with recurring epidemics. However, ongoing control attempts are being critically hampered by the lack of up-to-date information regarding the vectors. To overcome this problem, we examined the current situation and distribution of dengue vectors on the island. Residences throughout the urban, suburban, and rural areas were inspected through wet and dry seasons between February 2009 and February 2010. Two vectors were encountered in the survey, with Aedes aegypti present in especially high numbers mostly in urban areas. Similar observations were noted for Ae. albopictus in rural areas. The former species was more abundant in outdoor containers, while the latter showed almost equivalent abundance both outdoors and indoors. The dengue virus was active in both urban and rural areas, and the number of cases of infection was higher in areas where Ae. aegypti was predominant. The abundance of immature Ae. albopictus was positively correlated with rainfall (r2 = 0.461; P < 0.05), but this was not the case for Ae. aegypti. For both species, the size of immature populations tended to increase with increasing intensity of rain, but heavy rains resulted in population loss. In addition to updating data regarding the larval habitats and locations (outdoors and indoors), this study highlighted the importance of spatial vector control stratification, which has the potential to reduce costs in control programs.
A cryptic Bornean torrent frog of the genus Meristogenys, which is divergent genetically and morphologically from all known congeners, is described from mountain streams of western Sarawak, East Malaysia (Borneo). The species occurs sympatrically with the type species of the genus, M. jerboa, but apparently differs from it in adult coloration and larval morphology, such as keratodont formulae and glands in tail fins. Females of the new species possess much larger and fewer eggs than in sympatric M. jerboa, suggesting significantly different reproductive traits between these species. A key to larvae of known species of the genus is provided.
A new species of black fly, Simulium (Gomphostilbia) rampae, is described, based on adult male, its pupal exuviae and mature larvae collected from Doi Inthanon National Park, northern Thailand. This new species is placed in the Simulium asakoae species-group, and is characterized in the male by the high number of upper-eye facets in 17 vertical columns and 18 horizontal rows, in the pupa by the gill with a long common basal stalk, cone-shaped terminal hook, and cocoon with an anterodorsal projection, and in the larva by the medium-long postgenal cleft. A DNA analysis using COI gene supported its assignment to the S. asakoae species-group and showed its close relationship to S. (G.) udomi Takaoka & Choochote and S. (G.) chiangdaoense Takaoka & Srisuka. This is the fourth member of the S. asakoae species-group recorded from Thailand.
The final instar larva of Orthetrum borneense Kimmins, 1936, is described and figured for the first time based on exuviae from three male and six female larvae collected in Sarawak, Borneo (East Malaysia). It is compared with an early instar larva, which was matched to the adult O. borneense by DNA barcoding, and the known larvae of other species of this genus that occur in the region.