Cx. quinquefasciatus is a common nuisance mosquito widely distributed in tropical and subtropical areas. This mosquito is also a vector of urban filariasis. Control with chemicals has been hampered by the development of resistance against chemical insecticides and rising problems of environmental contamination associated with them. Therefore, it is important to adopt more integrated mosquito management approaches that include sustainable, non chemical solutions. The mermithid nematode Romanomermis iyengari is one of several natural control alternatives to synthetic pesticides for mosquito suppression. This study evaluated the effectiveness of the nematode R. iyengari for control of Cx. quinquefasciatus. The nematode R. iyengari was mass-produced, and pre-parasitics (J2) were used for laboratory and field experiments. In laboratory experiments, two concentrations of pre-parasitics (5 and 10 J2 per larva) were tested against L1, L2 and L3 instars larvae of Cx. quinquefasciatus. Infected larvae were observed daily to determine their mortality rate and the number of postparasitic nematodes emerging from dead larvae. In field experiments, 1000, 2000 and 3000 J2/m2 were sprayed in separate natural Cx. quinquefasciatus breeding sites. After treatment, the larval mosquito density in the breeding sites was assessed every 5 days. Laboratory results showed that all tested Cx. quinquefasciatus instars larvae were susceptible to nematode infection. The mortality rates observed for each larval stage indicated that the concentration of 10 J2 kills larvae faster, and that the L1 larvae died earlier than older larvae. The average number of post-parasitic nematodes emerging per larva increases with increasing nematode concentration; also more post-parasitic nematodes emerged from the L2 larvae. Field data showed that, in breeding site treated with 3000 J2 per square meter, larval mosquito reduction reached 97% after nematode application. The dosage of 1000 J2 per square meter did not reduce the larval density. The insect parasitic nematode R. iyengari could be easily used as component of integrated mosquitoes control program in lymphatic filariasis endemic countries.
Chronic exposure to mercury (Hg) can lead to cumulative impairments in motor and cognitive functions including alteration in anxiety responses. Although several risk factors have been identified in recent year, little is known about the environmental factors that either due exposure toward low level of inorganic mercury that may led to the developmental disorders. The present study investigated the effects of embryonic exposure of mercury chloride on motor function and anxiety-like behavior. The embryo exposed to 6 different concentrations of HgCl2 (7.5, 15, 30, 100, 125, 250nM) at 5hpf until hatching (72hpf) in a semi-static condition. The mortality rate increased in a dose dependent manner where the chronic embryonic exposure to 100nM decreased the number of tail coiling, heartbeat, and swimming activity. Aversive stimulus was used to examine the effects of 100nM interferes with the development of anxiety-related behavior. No elevation in both thigmotaxis and avoidance response of 6dpf larvae exposed with 100nM were found. Biochemical analysis showed HgCl2 exposure affects proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. These results showed that implication of HgCl2 on locomotor and biochemical defects affects motor performance and anxiety-like responses. Yet, the potential underlying mechanisms these responses need to be further investigated which is crucial to prevent potential hazards on the developing organism due to neurotoxicant exposure.
The control of mosquitoes is threatened by the appearance of insecticide resistance and therefore new control chemicals are urgently required. Here we show that inhibitors of mosquito peptidyl dipeptidase, a peptidase related to mammalian angiotensin-converting enzyme (ACE), are insecticidal to larvae of the mosquitoes, Aedes aegypti and Anopheles gambiae. ACE inhibitors (captopril, fosinopril and fosinoprilat) and two peptides (trypsin-modulating oostatic factor/TMOF and a bradykinin-potentiating peptide, BPP-12b) were all inhibitors of the larval ACE activity of both mosquitoes. Two inhibitors, captopril and fosinopril (a pro-drug ester of fosinoprilat), were tested for larvicidal activity. Within 24 h captopril had killed >90% of the early instars of both species with 3rd instars showing greater resistance. Mortality was also high within 24 h of exposure of 1st, 2nd and 3rd instars of An. gambiae to fosinopril. Fosinopril was also toxic to Ae. aegypti larvae, although the 1st instars appeared to be less susceptible to this pro-drug even after 72 h exposure. Homology models of the larval An. gambiae ACE proteins (AnoACE2 and AnoACE3) reveal structural differences compared to human ACE, suggesting that structure-based drug design offers a fruitful approach to the development of selective inhibitors of mosquito ACE enzymes as novel larvicides.
Effects of laterite cover soil with different characteristics on survival of buried eggs, third instar larvae, and pupae of Musca domestica (L.) were studied experimentally. Soil treatments were loose dry soil, loose wet soil, compacted dry soil, and compacted wet soil (CWS). Eggs, third instar larvae, and pupae were buried under 30 cm of the different soil treatments and placed under field conditions until adults emerged. Rearing medium was provided for eggs and larvae, and control treatments of all stages were unburied immatures placed on soil surface. Egg and pupal survival to adult were significantly affected by the cover soil treatments, but third instars were more resilient. Wet soil treatments (loose wet soil and CWS) resulted in significantly reduced pupal survival, but increased survival of eggs. However, CWS significantly reduced adult emergence from buried eggs. Though emergence of house flies buried as eggs was significantly reduced, some were able to hatch and emerging first instar larvae developed to pupation. Although cover soil does not completely prevent fly emergence, it did limit development and emergence of buried house flies.
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 increasing attention on Vietnam as a biodiversity hotspot prompted an investigation of the potential for cryptic diversity in black flies, a group well known elsewhere for its high frequency of isomorphic species. We analyzed the banding structure of the larval polytene chromosomes in the Simulium tuberosum species group to probe for diversity beyond the morphological level. Among 272 larvae, 88 different chromosomal rearrangements, primarily paracentric inversions, were discovered in addition to 25 already known in the basic sequences of the group in Asia. Chromosomal diversity in Vietnam far exceeds that known for the group in Thailand, with only about 5% of the rearrangements shared between the two countries. Fifteen cytoforms and nine morphoforms were revealed among six nominal species in Vietnam. Chromosomal evidence, combined with available molecular and morphological evidence, conservatively suggests that at least five of the cytoforms are valid species, two of which require formal names. The total chromosomal rearrangements and species (15) now known from the group in Vietnam far exceed those of any other area of comparable size in the world, supporting the country's status as a biodiversity hotspot. Phylogenetic inference based on uniquely shared, derived chromosomal rearrangements supports the clustering of cytoforms into two primary lineages, the Simulium tani complex and the Southeast Asian Simulium tuberosum subgroup. Some of these taxa could be threatened by habitat destruction, given their restricted geographical distributions and the expanding human population of Vietnam.
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.
Teleosts are unique among vertebrates due to their indeterminate muscle growth, i.e., continued production of neonatal muscle fibers until death. However, the molecular mechanism(s) underlying this property is unknown. Here, we focused on the torafugu (Takifugu rubripes) myosin heavy chain gene, MYHM2528-1, which is specifically expressed in neonatal muscle fibers produced by indeterminate muscle growth. We examined the flanking region of MYHM2528-1 through an in vivo reporter assay using zebrafish (Danio rerio) and identified a 2100 bp 5'-flanking sequence that contained sufficient promoter activity to allow specific gene expression. The effects of enhanced promoter activity were observed at the outer region of the fast muscle and the dorsal edge of slow muscle in zebrafish larvae. At the juvenile stage, the promoter was specifically activated in small diameter muscle fibers scattered throughout fast muscle and in slow muscle near the septum separating slow and fast muscles. This spatio-temporal promoter activity overlapped with known myogenic zones involved in teleost indeterminate muscle growth. A deletion mutant analysis revealed that the -2100 to -600 bp 5'flanking sequence of MYHM2528-1 is essential for promoter activity. This region contains putative binding sites for several representative myogenesis-related transcription factors and nuclear factor of activated T-cell (NFAT), a transcription activator involved in regeneration of mammalian adult skeletal muscle. A significant reduction in the promoter activity of the MYHM2528-1 deletion constructs was observed in accordance with a reduction in the number of these binding sites, suggesting the involvement of specific transcription factors in indeterminate muscle growth.
Although indigenous climbing perch (Anabas testudineusis) is a highly valuable species, slow growth pattern during the culture period impeding its commercial success in aquaculture. In many fish species, it has been demonstrated that incubation temperature of eggs influenced the muscle development and growth rates, which persisted throughout the subsequent larval and juvenile phases. Therefore, this study aimed to investigate whether different incubation temperature of eggs prior to hatching can stimulate the muscle development, growth, and growth-related gene expression of the slow-growing indigenous species of climbing perch. The fertilized eggs of A. testudineus from an artificial breeding program were incubated under control temperature of 24 °C (IT24), 26 °C (IT26), 28 °C (IT28), and 30 °C (IT30) in 10L glass aquaria with four replicated units for each temperature treatment. After hatching, the larvae from each incubated temperature were separately reared at ambient temperature for 10 days in aquarium, 20 days in hapas, and the next 42 days in cages, totaling 72 days post-hatching (dph). The hatching rates were found significantly (P
Natural insecticides from plant origin against mosquito vectors have been the main concern for research due to their high level of eco-safety. Control of mosquitoes in their larval stages are an ideal method since Aedes larvae are aquatic, thus it is easier to deal with them in this habitat. The present study was specifically conducted to explore the larvicidal efficacy of different plant parts of Ipomoea cairica (L.) or railway creeper crude extract obtained using two different solvents; methanol and acetone against late third-stage larvae of Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae). Plant materials of I. cairica leaf, flower, and stem were segregated, airdried, powdered, and extracted using Soxhlet apparatus. Larvicidal bioassays were performed by using World Health Organization standard larval susceptibility test method for each species which were conducted separately for different concentration ranging from 10 to 450 ppm. Both acetone and methanol extracts showed 100% mortality at highest concentration tested (450 ppm) after 24 h of exposure. Results from factorial ANOVA indicated that there were significant differences in larvicidal effects between mosquito species, solvent used and plant parts (F=5.71, df=2, P<0.05). The acetone extract of I. cairica leaf showed the most effective larvicidal action in Ae. aegypti with LC50 of 101.94 ppm followed by Ae. albopictus with LC50 of 105.59 ppm compared with other fractions of I. cairica extract obtained from flower, stem, and when methanol are used as solvent. The larvae of Ae. aegypti appeared to be more susceptible to I. cairica extract with lower LC50 value compared with Ae. albopictus (F=8.83, df=1, P<0.05). Therefore, this study suggests that the acetone extract of I. cairica leaf can be considered as plant-derived insecticide for the control of Aedes mosquitoes. This study quantified the larvicidal property of I. cairica extract, providing information on lethal concentration that may have potential for a more eco-friendly Aedes mosquito control program.
A study on the chironomids (Diptera:Chironomidae) diversity at pristine ecosystem was carried out at upstream of Sungai Langat, Selangor. The study determines chironomids distribution and composition at 7 streams and river within the upstream site of Langat Catchment. Chironomid was sampled using Surber net and water quality was measured based on Malaysia WQI. The result indicated that Chironomidae was represented by three subfamilies namely Chironominae, Orthocladiinae and Tanypodinae, which comprises of 2502 individuals. Chironominae was the most dominant subfamily (1619 individuals) followed by Orthocladinae (629 individuals) and Tanypodinae (254 individuals). Polypedilum (subfamily: Chironominae) is the most dominant genus found followed by Rheocricotopus (subfamily: Orthocladiinae), Microtendipes and Cryptochironomus. Polypedilum was abundant in all stations except Sg. Langat 3 which was dominated by Rheocricotopus. Sungai Langat 3 has the highest number of individual (1113) which is (44.5%) from total chironomid individual and followed by Sg. Lolo with 468 individuals that were dominated by Polypedilum.
Hypopygiopsis violacea, a species of fly of forensic importance, was recovered from a corpse and described for the first time. The morphological structures of the second and third instar larvae of four specimens were examined using light microscope. Observations were focused on three main morphological characters: cephalopharyngeal skeleton, anterior and posterior spiracles. Cephalopharyngeal skeleton of second instar larva is darkly pigmented and without accessory sclerite below the mouth hook. The anterior spiracles of second and third instar larvae have 8-9 papillae each, arranged in a single row. The posterior spiracle of second instar larva has two spiracular slits with no thickening of peritreme. This differentiates it from the third instar, whereby the latter has three slits for each posterior spiracle. Cephalopharyngeal skeleton of third instar larva is heavily pigmented. An accessory sclerite is found below the hook part of third instar larva but is absent in second instar. Peritreme of the posterior spiracle of third instar larva is thick almost complete encircling a button. The intersegmental spines of the cuticular surface are dome-shaped and unicuspid. Third instar larva of this species is large with size approximately 15 mm long. These findings provide important identification features of immature stages of Hy. violacea which could be useful in forensic entomology.
Chrysomya megacephala (Fabricius) dan Chrysomya rufifacies (Macquart) adalah merupakan dua spesies lalat penting yang boleh dijadikan sebagai penunjuk terbaik untuk menganggarkan selang masa kematian atau Post Mortem Interval (PMI) untuk kegunaan dalam sains forensik. Penentuan PMI adalah berdasarkan kepada saiz dan peringkat perkembangan larva. Kajian perkembangan telur, larva dan pupa lalat Ch. megacepahala dan Ch. rufifacies dijalankan di bawah suhu 27ºC, 30ºC dan 33ºC di makmal dengan menggunakan kebuk pertumbuhann serangga. Data daripada kajian digunakan untuk menghasilkan satu graf pertumbuhan dan Jam Darjah Terkumpul (ADH) bagi kedua-dua spesies. Ch. megacephala mengambil masa selama 9.15 hari pada suhu 27ºC, 8.54 hari (30ºC) dan 6.76 hari (33ºC) untuk melengkapkan satu kitar hidup. Pada C. rufifacies pula, kitar hidupnya lebih lama berbanding Ch. megacephala iaitu 9.92 hari pada suhu 27ºC, 9.13 hari (30ºC) dan 7.44 hari (33ºC). Telur bagi kedua-dua spesies menetas lebih cepat pada suhu 33ºC berbanding dua suhu yang lainnya. Nilai ADH yang rendah pada sesuatu suhu, menunjukkan sesuatu spesies lebih cepat melengkapkan suatu kitar hidup. Penemuan ini berguna dalam menganggarkan selang masa kematian bagi mayat yang dijumpai pada suhu persekitaran yang berlainan.
Megaselia scalaris (Loew) is one of the best-known diets for the swiftlet. Previous studies have addressed the problem of some mass rearing conditions for this insect; unfortunately, the details of the nutritional composition of the life stages and cost of the breeding materials were insufficiently reported, even though this information is crucial for farming the edible-nest swiftlet. We aimed to investigate the nutritional composition of the life stages of M scalaris on a cost basis using 3 common commercial breeding materials: chicken pellets (CPs), fish pellets (FPs), and mouse pellets (MPs). Modified Association of Official Analytical Chemists (AOAC) proximate and mineral analyses were carried out on the insect's third instar larvae, pupal, and adult stages to determine the nutritional composition. Regardless of the breeding materials, the adult stage of M scalaris had significantly higher crude protein than the other stages; the pupae were rich in calcium, which is required for egg production; and the third instar larvae had the highest amount of crude fat compared with the other stages. Regarding the energy content, there were no significant differences among the stages according to the breeding materials. In terms of nutritional cost, CP was the most economic breeding material and generated the highest amount of nutrients per US dollar (US $). Different life stages of M scalaris were used by the swiftlets by supplying the required nutrients, and future studies should focus on effective diet feeding methods.
Myiasis is a pathological condition in humans and animals caused by various species of dipterous larvae. Myiasis which occurs in a newborn baby is referred as neonatal myiasis. It is a rare condition and there are only a few reports to date. A case of neonatal aural myiasis in a two day old infant is reported in this paper.
Mosquito adults and larvae were collected from dengue high risk areas and transported to the laboratory for identification. Identified mosquitos were pooled according to the species, date and locality and stored at -70 degrees C. A total of 1,385 pools of Aedes albopictus and 267 pools of Ae. Aegypti were collected from major towns in 12 states in Peninsular Malaysia. Virus isolation was carried out using cell culture (C6/36 clone) of Ae. albopictus and detection of dengue virus by the peroxidase anti-peroxidase staining. All positive isolations were further re-confirmed by the reverse transcriptase-polymerase chain reaction (RT-PCR). Most of the pools were negative with PAP staining and RT-PCR. However, 11 mosquito pools were positive with PAP staining. On the other hand, samples from Terengganu, Pulau Pinang and Johor were positive using both methods.
The effect of ten microalgal chlorophytes isolated from mosquito breeding containers on the survival, larval development and adult body size of the mosquito Aedes aegypti was investigated. All larvae fed with six of the microalgal isolates died after 7 days. These isolates were found to be resistant to digestion by mosquito larvae. Delayed pupation and body size reduction of the mosquitos fed with Chlorococcum UMACC 218 and Scenedesmus UMACC 220 were observed. In contrast, larvae fed with Ankistrodesmus convolutus UMACC 101 and Chlorococcum UMACC 213 were bigger in size than those fed with normal insectory feed. The present study showed that microalgal chlorophytes have the potential to be used as larvicidal agents for mosquitos.
A large scale study was conducted to elucidate the true relationship among entomological, epidemiological and environmental factors that contributed to dengue outbreak in Malaysia. Two large areas (Selayang and Bandar Baru Bangi) were selected in this study based on five consecutive years of high dengue cases. Entomological data were collected using ovitraps where the number of larvae was used to reflect Aedes mosquito population size; followed by RT-PCR screening to detect and serotype dengue virus in mosquitoes. Notified cases, date of disease onset, and number and type of the interventions were used as epidemiological endpoint, while rainfall, temperature, relative humidity and air pollution index (API) were indicators for environmental data. The field study was conducted during 81 weeks of data collection. Correlation and Autoregressive Distributed Lag Model were used to determine the relationship. The study showed that, notified cases were indirectly related with the environmental data, but shifted one week, i.e. last 3 weeks positive PCR; last 4 weeks rainfall; last 3 weeks maximum relative humidity; last 3 weeks minimum and maximum temperature; and last 4 weeks air pollution index (API), respectively. Notified cases were also related with next week intervention, while conventional intervention only happened 4 weeks after larvae were found, indicating ample time for dengue transmission. Based on a significant relationship among the three factors (epidemiological, entomological and environmental), estimated Autoregressive Distributed Lag (ADL) model for both locations produced high accuracy 84.9% for Selayang and 84.1% for Bandar Baru Bangi in predicting the actual notified cases. Hence, such model can be used in forestalling dengue outbreak and acts as an early warning system. The existence of relationships among the entomological, epidemiological and environmental factors can be used to build an early warning system for the prediction of dengue outbreak so that preventive interventions can be taken early to avert the outbreaks.
Natural peptides in insect vectors played an important role in the control of
pathogens. Musca domestica Linnaeus and Chrysomya megacephala Fabricius were
two species of local fly chosen to detect presence of antimicrobial peptide substance.
The screening of the antimicrobial activity was carried using a spectrophotometric
method. Results were obtained much quicker and less laborious. The results showed
larva hemolymph of M. domestica lysed Bacillus subtilis and two Gram negatives,
Escherichia coli and Pseudomona. aeruginosa. The pupae hemolymph only lysed E.
coli. Whereas, the hemolymph of C. megacephala larva showed bactericidal effect
against both of the Gram positives tested, i.e. B. subtilis and Staph. aureus. and no
effect was against the Gram negatives. The pupa showed lytic activity against Staph.
aureus and P. aeruginosa. As a conclusion, the larva and pupa hemolymph of M.
domestica and C. megacephala demonstrated antibacterial activity. However, larva
hemolymph of M. domestica and C. megacephala has broader antibacterial activity
against both Gram positive and negative bacteria.