In forensic entomology, breeding of fly larvae in a controlled laboratory environment using animal tissue is a common technique to obtain insect developmental time for the estimation of postmortem interval. Previous studies on growth media are mostly on the effect of different diets on fly development. However, the interaction effects between temperature and food type used have not been explored. The objective of this study was to compare the use of cow's liver agar and raw liver on the development of a forensically important fly, Megaselia scalaris (Loew). This study also determined the interaction between different temperatures and different food types on the growth of this species. A total of 100 M. scalaris eggs were transferred into each of the two media mentioned above. Liver agar was prepared by adding dried ground liver into nutrient agar, whilst raw liver was naturally prepared from the same animal source. This experiment was conducted at 27, 30 and 33 °C in an incubator in a continuously dark condition. Length and weight of larvae, puparia and adult samples were determined. Total developmental times for larvae feeding on liver agar at each temperature were approximately 7-15 h slower than those feeding on raw liver. Survival rates were almost equal in both diets but were lower at 33 °C. Mean larva length in both diets did not differ significantly at all temperatures, but larvae feeding on liver agar had lower mean weight values than those in raw liver at 30 and 33 °C. The effect of temperature was significant in female puparia weight and male adult weight whereas the effect of diet types was significant in both male and female puparia size and weight. Interaction effects of temperature and food type on M. scalaris puparium size and adult weight were significant, indicating that puparium size and adult weight depended on both food type and temperature. This experiment highlighted the use of cow's liver agar as an alternative diet to breed M. scalaris in the laboratory and the importance of considering the interaction effect between temperatures and food types when deciding the most suitable medium in fly larva rearing.
Methods are described for the laboratory colonization of Mansonia uniformis, Ma. indiana and Ma. bonneae in Malaysia. Gravid females oviposited in 500 ml beakers with a layer of water covered with small leaves of Salvinia. Newly hatched larvae were set up in a basal medium of guinea pig dung and water or liver powder, yeast powder and water. Larvae attached to aquatic plants or 'Keaykolour' ruffia snow white paper. The cultures with paper gave better yields than those with plants. Production of Ma. uniformis was higher than the other two species. Twelve generations of Ma. uniformis and 11 generations of Ma. indiana and Ma. bonneae were monitored in the laboratory.
A burned human remain was found outdoor (5º 27' N, 100º 16' E) in Penang Island. The deceased was last seen alive on 23 April 2010 at 2230 h and was found burned on 24 April 2010 at 1920 h. Larval aggregation of second instar Chrysomya megacephala was observed on the chest of the deceased.
Detailed studies of larval development of Octolasmis angulata and Octolasmis cor are pivotal in understanding the larval morphological evolution as well as enhancing the functional ecology. Six planktotrophic naupliar stages and one non-feeding cyprid stage are documented in details for the first time for the two species of Octolasmis. Morphologically, the larvae of O. angulata and O. cor are similar in body size, setation patterns on the naupliar appendages, labrum, dorsal setae-pores, frontal horns, cyprid carapace, fronto-lateral gland pores, and lattice organs. Numbers of peculiarities were observed on the gnathobases of the antennae and mandible throughout the naupliar life-cycle. The setation pattern on the naupliar appendages are classified based on the segmentation on the naupliar appendages. The nauplius VI of both species undergoes a conspicuous change before metamorphosis into cyprid stage. The cyprid structures begin to form and modify beneath the naupliar body towards the end of stage VI. This study emphasises the importance of the pedunculate barnacle larval developmental studies not only to comprehend the larval morphological evolution but also to fill in the gaps in understanding the modification of the naupliar structures to adapt into the cyprid life-style.
The larval growth of Liosarcophaga dux Thompson (Diptera: Sarcophagidae) was studied under varying indoor room temperatures in Malaysia. Five replicates were established. The immature growth of this species from first instar until adult emergence was 307.0+/-3.0 hours. The mean larval length measured for second instar, third instar, post-feeding stage and puparia were 6.5+/-0.5 mm (n=10), 11.8+/-3.7 mm (n=31), 12.7+/-0.8 mm (n=16), and 9.5+/-0.5 mm (n=15), respectively.
Larvae of the Synthesiomyia nudiseta (Wulp) were collected from a decomposed human corpse at the Department of Forensic Medicine, Penang Hospital. A colony of this species was established and the eggs were collected for rearing. The developmental times, rearing temperatures, and relative humidity were recorded twice daily from the time the eggs collected until adult emergence. An average of 5 larvae were randomly collected from the rearings twice daily, warm-water killed and preserved in Kahle's solution. The larval instar stages were determined by observing the number of posterior spiracular slits and the length of the preserved larvae were measured. When the larval life cycle was completed, the accumulated developmental times were calculated. A total of 8 replicates were carried out. The temperature of the rearing room was 28.5+/-1.5 degrees Celcius while the relative humidity was within 67-85%. The total developmental time for S. nudiseta was 322+/-19 hours (13.4+/-0.8 days).
The presence of predators can have dramatic consequences on prey communities, not only by the direct effects of consumption but also through sublethal effects. We investigated the survival rate and subsequent life history of the mosquito Culex pervigilans Bergroth under the influence of its major predator, the backswimmer Anisops wakefieldi White. We established a field experiment with various treatments: 1) control without predators, 2) free-roaming A. wakefieldi (with one, three, or nine A. wakefieldi per container), 3) caged A. wakefieldi (empty cage without predators, with one, three, or nine A. wakefieldi in each cage, and 4) A. wakefieldi cues (with cues concentrations of one, three, or nine A. wakefieldi). Cx. pervigilans eggs were then taken from these four experimental treatments and reared in two different laboratory conditions: 1) in clean water without any traces of predators, or 2) in water with the same treatments as in field. The survival rate of Cx. pervigilans was significantly reduced by the presence of predators or their cues. Even after a brief exposure to waters containing predators or residual cues, the subsequent progeny and the ontogeny of the remaining survivors were still affected. The percentage of eggs that hatched, and the resulting mosquito population, was influenced by the presence of predators or their cues. Our results suggest that sublethal effects may be carried by surviving individuals primarily through the effects of stress, perhaps by epigenetic mechanisms. We may expect to observe similar plasticity in species or populations with high temporal or spatial variability in predation.
Life tables were constructed for twelve cohorts of immature stages of the dengue vector Ae. albopictus in a wooded area of Penang, Malaysia. The development time of Ae. albopictus ranged from 6 to 10 days depending on the mean environmental temperature (r = - 0.639, p < 0.05). Total cohort mortality was correlated with total development time (r = 0.713, p < 0.05) but not temperature (r = -0.477, p > 0.05). Rainfall was correlated with neither development time (r = 0.554, p > 0.05) nor mortality (r = 0.322, p > 0.05). There was a significant difference among the total mortality that occurred in the twelve cohorts (H = 119.783, df = 11, p < 0.05). There was also a significant difference in mortality among the different stages (H = 274.00, df = 4, p < 0.05).
In the present study on the life-cycle of Blomia tropicalis, freshly laid eggs were observed until they developed into adults; the development periods between stages were recorded. The eggs took an average of 22.9 +/- 6.4 days to develop to adults. For longevity experiments, newly emerged adults were kept at 25 degrees C and observed until they died. There was no significant difference in longevities of the different sexes (p = 0.053). Production of eggs by mated females were monitored until egg production stopped and the female died. Mated females and males survived an average of 32.2 +/- 15.4 and 30.9 +/- 17.7 days respectively. The difference in longevity of the mated females, and males was not significant (p = 0.747). Longevity of the mated females was found to be significantly (p < 0.05) shorter than unmated females.
The use of Chrysomya megacephala larvae for detecting malathion for diagnosing the cause of death was investigated. This could prove useful when the visceral organs have become liquefied during decomposition and therefore cannot be sampled. A field experiment was conducted in which C. megacephala were allowed to colonise naturally the corpses of rabbits that had died of malathion poisoning. The concentration of malathion increased gradually during the larval stages of C. megacephala reaching the maximum concentration in the third instar larvae. The concentration of malathion declined during prepupal stage and reached its lowest level among tenerals. The average malathion concentrations in C. megacephala growing in poisoned rabbit corpses left in a sunlit habitat were significantly higher (p<0.05) than those growing on poisoned rabbits left in a shaded habitat. The concentrations of malathion in the different stages of development of C. megacephala were moderately correlated (r = 0.51-0.69) with the administered doses as well as with those estimated in visceral organs. Thus, it would not be reliable to suggest the formulation of mathematical algorithms for relating the concentration of malathion found in the different stages of development of C. megacephala with those found in the visceral organs. However, in the context of forensic investigation, the qualitative detection of malathion in C. megacephala may prove useful in diagnosing the cause of death, since malathion is a common cause of accidental and suicidal deaths.
Pieris rapae is a serious pest of brassicas worldwide. We performed de novo assembly of P. rapae transcriptome by next-generation sequencing and assembled approximately 65,727,422 clean paired-end reads into 32,118 unigenes, of which 13,585 were mapped to 255 pathways in the KEGG database. A total of 6173 novel transcripts were identified from reads directly mapped to P. rapae genome. Additionally, 1490 SSRs, 301,377 SNPs, and 29,284 InDels were identified as potential molecular markers to explore polymorphism within P. rapae populations. We screened and mapped 36 transcripts related to OBP, CSP, SNMP, PBAN, and OR. We analyzed the expression profiles of 7 selected genes involved in pheromone transport and degradation by quantitative real-time PCR; these genes are sex-specific and differentially expressed in the developmental stages. Overall, the comprehensive transcriptome resources described in this study could help understand and identify molecular targets particularly reproduction-related genes for developing effective P. rapae management tools.
We screened more than 60 Malaysian plants against two species of insects and found that Melicope subunifoliolata (Stapf) T.G. Hartley (Rutaceae) showed strong feeding deterrent activity against Sitophilus zeamais Motsch. (Curculionidae) and very good larvicidal activity against Aedes aegypti L. (Diptera). One anti-insect compound, meliternatin (3,5-dimethoxy-3',4',6,7-bismethylendioxyflavone) (6) and six other minor polyoxygenated flavones were isolated from M. subunifoliolata.
Previously, we reported the presence of imported trichinellosis in a Thai worker returning from Malaysia, who presented with progressive generalized muscle hypertrophy and weakness after eating wild boar meat. This work analyzed a partial small subunit of a mitochondrial ribosomal RNA gene of Trichinella larvae isolated from the patient. The results showed complete identity with a mitochondrial RNA gene of Trichinella papuae (GenBank accession no. EF517130). This is the first report of imported trichinellosis in Thailand caused by T. papuae. It is possible that T. papuae is widely distributed in the wildlife of Southeast Asia.
In addition to causing myiasis in humans, Megaselia spiracularis Schmitz has also been reported as a forensically important fly. In this study, we presented the morphology of all larval instars and puparium of M. spiracularis using scanning electron microscopy. The first instar larva was composed of 12 segments. Antennae and maxillary palp complex were visible. Two spiracular slits could be seen at the posterior spiracle. The branch of posterior spiracular hairs was approximately equal to the palm-formed base in length. Second and third larval instars were very similar to first instar, except for the presence of anterior spiracle. The labium of the second instar larva was triangular and ventrally curved, whereas it was a bilobed structure and the tip forked in the first instar. The bubble membrance comprised of ≈40 globules presented at the third instar larvae. Puparia showed a retracted cephalic region and a pair of pupal respiratory horns on the dorsum. A comparison of the morphological features between immature stages of M. spiracularis and M. scalaris, a forensically important fly indoors in Germany, Malaysia, and China, was discussed.
Four new species of black flies are described, and three others are reported as newly recorded, based on adults reared from pupae, pupae and larvae collected in and near Tam Dao National Park, Vinh Phuc Province, Vietnam. New species include Simulium (Gomphostilbia) hongthaii sp. nov., S. (G.) tamdaoense sp. nov. (both species placed in the asakoae species-group), S. (Simulium) taythienense sp. nov. and S. (S.) xuandai sp. nov. (the two latter species placed in the striatum species-group). Newly recorded species are S. (G.) brinchangense Takaoka, Sofian-Azirun & Hashim, S. (Nevermannia) aureohirtum Brunetti and S. (S.) brevipar Takaoka & Davies. These discoveries increase the number of species of black flies known in Vietnam from 21 to 28.
Even with continuous vector control, dengue is still a growing threat to public health in Southeast Asia. Main causes comprise difficulties in identifying productive breeding sites and inappropriate targeted chemical interventions. In this region, rural families keep live birds in backyards and dengue mosquitoes have been reported in containers in the cages. To focus on this particular breeding site, we examined the capacity of bird fecal matter (BFM) from the spotted dove, to support Aedes albopictus larval growth. The impact of BFM larval uptake on some adult fitness traits influencing vectorial capacity was also investigated. In serial bioassays involving a high and low larval density (HD and LD), BFM and larval standard food (LSF) affected differently larval development. At HD, development was longer in the BFM environment. There were no appreciable mortality differences between the two treatments, which resulted in similar pupation and adult emergence successes. BFM treatment produced a better gender balance. There were comparable levels of blood uptake and egg production in BFM and LSF females at LD; that was not the case for the HD one, which resulted in bigger adults. BFM and LSF females displayed equivalent lifespans; in males, this parameter was shorter in those derived from the BFM/LD treatment. Taken together these results suggest that bird defecations successfully support the development of Ae. albopictus. Due to their cryptic aspects, containers used to supply water to encaged birds may not have been targeted by chemical interventions.
Nectar is the staple diet of adult mosquitoes in the wild, but its availability is inconsistent and can be affected by rainfall. In urban centers, Aedes vectors commonly use man-made containers as their major habitat; however, they can colonize any items replenished by rainfall. Garbage output has increased significantly in recent years, at a time when collection frequency is reducing. Such garbage usually includes organic components, some of which are sweet and can be fed upon by other animals or become can containers for rainwater. Despite evidence that Aedes larvae can thrive in containers comprised of organic waste material, which can be produced by rodents gnawing on fruits or vegetables, and that adults can survive on sweet waste fluids, the capacity of organic waste materials to accumulate rainwater and act as egg deposition sites has not been examined. It is also unknown for how long sweet extracts can sustain the life of adult vectors. Here, we investigated the abundance of sweet leftovers at garbage sites and the rainwater retention capacity of some organic materials through a field survey and laboratory bioassays. We also examined whether sweet waste fluids impact egg hatching success and longevity of Aedes aegypti. The results of this study indicated that sweet products with leftovers are highly prevalent in garbage. When exposed to rain, food items (BAFrc, banana fruit resembling container; and BSPrc, boiled sweet potato resembling container) and the packaging of sweet foods (SMIc, sweetened condensed milk can) retained water. When provided an opportunity to oviposit in cups containing BAF extract (BAFex), BSP extract (BSPex), and SMI extract (SMIex), eggs were deposited in all media. Egg maturation in the BAFex environment resulted in similar larval eclosion success to that resulting from embryo development in a water milieu. Adults maintained on sweet waste extracts had long lifespans, although shorter than that of their sugar solution (SUS)-fed counterparts. Taken together, these results indicated that sweet waste materials are useful to dengue mosquitoes, acting both as oviposition sites and energy sources.