Field studies integrating pollination investigations with an assessment of floral scent composition and thermogenesis in tropical aroids are rather few. Thus, this study aimed to investigate the pollination biology of nine species belonging to Schismatoglottis Calyptrata Complex Clade. The flowering mechanism, visiting insect activities, reproductive system, thermogenesis and floral scent composition were examined. Anthesis for all species started at dawn and lasted 25-29 h. Colocasiomyia (Diptera, Drosophilidae) are considered the main pollinators for all the investigated species. Cycreon (Coleoptera, Hydrophilidae) are considered secondary pollinators as they are only present in seven of the nine host plants, despite the fact that they are the most effective pollen carrier, carrying up to 15 times more pollen grains than Colocasiomyia flies. However, the number of Colocasiomyia individuals was six times higher than Cycreon beetles. Chaloenus (Chrysomelidae, Galeuricinae) appeared to be an inadvertent pollinator. Atheta (Coleoptera, Staphylinidae) is considered a floral visitor in most investigated species of the Calyptrata Complex Clade in Sarawak, but a possible pollinator in S. muluensis. Chironomidae midges and pteromalid wasps are considered visitors in S. calyptrata. Thermogenesis in a biphasic pattern was observed in inflorescences of S. adducta, S. calyptrata, S. giamensis, S. pseudoniahensis and S. roh. The first peak occurred during pistillate anthesis; the second peak during staminate anthesis. Inflorescences of all investigated species of Calyptrata Complex Clade emitted four types of ester compound, with methyl ester-3-methyl-3-butenoic acid as a single major VOC (volatile organic compound). The appendix, pistillate zone, staminate zone and spathe emitted all these compounds. A mixed fly-beetle pollination system is considered an ancestral trait in the Calyptrata Complex Clade, persisting in Sarawak taxa, whereas the marked reduction of interpistillar staminodes in taxa from Peninsular Malaysia and especially, Ambon, Indonesia, is probably linked to a shift in these taxa to a fly-pollinated system.
The major fruit fly attractant component in the floral fragrance of Bulbophyllum cheiri (fruit fly orchid) is methyl eugenol (ME). In the lowland rain forest of Malaysia, the solitary and nonresupinate flowers of the fruit fly orchid attract only males of the ME-sensitive fruit fly species (Bactrocera carambolae, B. papayae. and B. umbrosa. During the morning, the fruit fly orchid flower is visited by many fruit flies, which can sometimes cover the whole flower. The number of visitors dwindles in the afternoon. Headspace analysis of the flower shows a high ME peak in the morning, a small one between 12:00 and 14:00 hr, and no detectable ME peak after 14:00 hr. The process of pollination in the wild is initiated by attraction of fruit flies to floral ME. The flower, with the aid of its specialized hinged see-saw lip (labellum), temporarily traps (< 1 min) a fruit fly pollinator between its lip and column. Just prior to this, the fly is rewarded by the opportunity to feed on the floral attractant found on surfaces of petals, sepals, and lip. The pollinaria borne by two wild B. papayae males (caught on and near the fruit fly orchid flower) are identical in morphology and structure with those obtained from the flower. Many of the B. papayae males (17 of 22 analyzed) attracted to the fruit fly orchid already possessed both ME metabolites, trans-coniferyl alcohol and 2-allyl-4,5-dimethoxyphenol, in their rectal glands. indicating that they had previously consumed ME. In this orchid-fruit fly association, both organisms gain direct reproductive benefits: the orchid flower gets pollinated without having to offer nectar, while the fruit fly boosts its pheromone and defense system, as well as its sexual competitiveness by feeding on the ME produced by the flower.
Methyl eugenol (ME), is converted into two major phenylpropanoids, 2-allyl-4,5-dimethoxyphenol and trans-coniferyl alcohol, following consumption by the male fruit fly Bactrocera papayae. Chemical analysis of wild male B. papayae rectal glands, where the compounds are sequestered, revealed the presence of ME metabolites in varying quantities. These phenylpropanoids are shown to be involved in the fruit fly defense both in no-choice and choice feeding tests against the Malayan spiny gecko, Gekko monarchus. After being acclimatized to feeding on fruit flies, geckos consumed significantly fewer ME-fed male flies than controls that consumed all the ME-deprived male flies offered throughout a two-week period. Diagnosis of dissected livers from geckos that consumed ME-fed male flies revealed various abnormalities. These included discoloration and hardening of liver tissue, whitening of the gallbladder, or presence of tumor-like growths in all geckos that consumed ME-fed male flies. Control geckos fed on ME-deprived male flies had healthy livers. When given an alternative prey, geckos preferred to eat untreated house flies, Musca domestica to avoid preying on ME-fed fruit flies.
The performance of newly developed trapping systems for the Old World screw-worm fly, Chrysomya bezziana has been determined in field trials on cattle farms in Malaysia. The efficacy of non-sticky traps and new attractants to trap C. bezziana and non-target flies was compared with the standard sticky trap and Swormlure. The optimal trap was a modified LuciTrap(®) with a new attractant mixture, Bezzilure-2. The LuciTrap/Bezzilure-2 caught on average 3.1 times more C. bezziana than the sticky trap with Swormlure (P<0.05) and provided selectivity for C. bezziana against Chrysomya megacephala and Chrysomya rufifacies with factors of 5.9 and 6.4, respectively. The LuciTrap also discriminates with factors of 90 and 3.6 against Hemipyrellia sp. and sarcophagid flesh flies respectively, compared to the sticky trap. The LuciTrap/Bezzilure-2 system is recommended for screwworm fly surveillance as it is more attractive and selective towards C. bezziana and provides flies of better quality for identification than the sticky trap.
Signal fly, Scholastes sp. (Diptera: Platystomatidae) was observed associated with animal carcasses in Malaysia. The first observation was on a monkey carcass, which was killed by using a handgun and immediately placed in a forested area in Gombak, Selangor while the second observation was on a pig that died of natural causes and whose carcass was placed in an oil palm plantation in Tanjung Sepat, Selangor. Both animal carcasses were visited by Scholastes sp. flies during the fresh decomposition period. However, the role Scholastes flies in the decomposition process remains unknown. In this paper, we report the occurrence of Scholastes sp. on animal carcasses in Malaysia for the first time.
Phylogenetic, genealogical and population relationships of Chrysomya bezziana, the Old World screwworm fly (OWSF), were inferred from DNA sequences of mitochondrial cytochrome b (cyt b), nuclear elongation factor-1α (EF-1α) and nuclear white eye colour (white), using sequences of Chrysomya megacephala and Chrysomya rufifacies as outgroups. Cyt b (717bp, 754 specimens), EF-1α (361bp, 256 specimens) and white (577bp, 242 specimens) were analysed from up to two African and nine Asian countries, including 10 Indonesian islands. We show that OWSF occurs as distinctive African and Asian lineages based on cyt b and white, and that there is a marked differentiation between Sumatran and Javan populations in Indonesia, supported by the genealogy and analysis of molecular variance of cyt b alone. Four cyt b sub-lineages are recognised in Asia: only 2.1 occurs on the Asian mainland, from Yemen to Peninsular Malaysia; only 2.2, 2.3 and 2.4 occur in central Indonesia; 2.4 predominates on New Guinea; and 2.1 co-occurs with others only on Sumatra in western Indonesia. This phylogeography and the genetic distances between cyt b haplotypes indicate pre-historic, natural dispersal of OWSF eastwards into Indonesia and other Malesian islands, followed by vicariant evolution in New Guinea and central Indonesia. OWSF is absent from Australia, where there is surveillance for importation or natural invasion. Judged by cyt b haplotype markers, there is currently little spread of OWSF across sea barriers, despite frequent shipments of Australian livestock through Indonesian seas to the Middle East Gulf region. These findings will inform plans for integrated pest management, which could be applied progressively, for example starting in East Nusa Tenggara (central Indonesia) where OWSF has regional cyt b markers, and progressing westwards to Java where any invasion from Sumatra is unlikely. Cyt b markers would help identify the source of any re-emergence in treated areas.
Measuring wild pollinator services in agricultural production is very important in the context of sustainable management. In this study, we estimated the contribution of native pollinators to mango fruit set production of two mango cultivars Mangifera indica (L). cv. 'Sala' and 'Chok Anan'. Visitation rates of pollinators on mango flowers and number of pollen grains adhering to their bodies determined pollinator efficiency for reproductive success of the crop. Chok Anan failed to produce any fruit set in the absence of pollinators. In natural condition, we found that Sala produced 4.8% fruit set per hermaphrodite flower while Chok Anan produced 3.1% per flower. Hand pollination tremendously increased fruit set of naturally pollinated flower for Sala (>100%), but only 33% for Chok Anan. Pollinator contribution to mango fruit set was estimated at 53% of total fruit set production. Our results highlighted the importance of insect pollinations in mango production. Large size flies Eristalinus spp. and Chrysomya spp. were found to be effective pollen carriers and visited more mango flowers compared with other flower visitors.
A number of species are affected by Sex-Ratio (SR) meiotic drive, a selfish genetic element located on the X-chromosome that causes dysfunction of Y-bearing sperm. SR is transmitted to up to 100% of offspring, causing extreme sex ratio bias. SR in several species is found in a stable polymorphism at a moderate frequency, suggesting there must be strong frequency-dependent selection resisting its spread. We investigate the effect of SR on female and male egg-to-adult viability in the Malaysian stalk-eyed fly, Teleopsis dalmanni. SR meiotic drive in this species is old, and appears to be broadly stable at a moderate (approx. 20%) frequency. We use large-scale controlled crosses to estimate the strength of selection acting against SR in female and male carriers. We find that SR reduces the egg-to-adult viability of both sexes. In females, homozygous females experience greater reduction in viability (sf = 0.242) and the deleterious effects of SR are additive (h = 0.511). The male deficit in viability (sm = 0.214) is not different from that in homozygous females. The evidence does not support the expectation that deleterious side effects of SR are recessive or sex-limited. We discuss how these reductions in egg-to-adult survival, as well as other forms of selection acting on SR, may maintain the SR polymorphism in this species.
Cosmopolitan scuttle fly, Megaselia scalaris (Loew) (Diptera: Phoridae) is one of the commonest forensic species recorded colonizing human corpse indoors and in concealed environment. The occurrence of this species in such environments provides a higher evidential value to assist estimation of postmortem interval (PMI) compared to other forensically important dipterans. However, developmental and size data of M. scalaris are still lacking and they are derived from a limited range of thermal values. The objective of this study is to develop the growth model of M. scalaris by emphasizing the size range of larvae and puparia at different constant temperatures. This species was reared in six replicates at eight varying constant temperatures ranging from 23 to 36 °C and cow's liver was provided as food source. Larvae and puparia were sampled at set time intervals and measured by their length and weight. Because interpretation of forensic entomological evidence is subject to application of different techniques, development of M. scalaris is expressed herein by using developmental table, length/morphological stage diagrams and linear/nonlinear estimation methods. From the findings, it is very important to highlight that sexual dimorphism of M. scalaris during post feeding larva and pupa stage could be observed based on size and developmental periods. Mean length and weight ratios of male to female puparia are approximately 0.8 and 0.3-0.5, respectively, indicating sexual dimorphism of this species. Developmental period in female are 4.0-11.4 h (post feeding larval stage), 3.7-24.0 h (pupal stage), and 3.0-20.1 h (total developmental period) longer in male. Due to this dimorphism, PMI estimation using M. scalaris post feeding larva or puparium specimens must be carried out carefully by to avoid inaccuracy and misinterpretation.
The study on biodiversity of forensically important Diptera in the tropical rain forest in Malaysia is scarce. Thus, a preliminary survey was conducted at a jungle fringe near Kampung Bahagia Bukit Lagong, Sungai Buloh, Selangor. A rat carcass was offered to attract carrion flies and we collected an adult female calliphorid, Hypopygiopsis fumipennis (Walker, 1856) during the fresh stage of carcass decomposition. The female fly was allowed to oviposit on chicken liver in a container and the resulting larvae were reared to the adult stage. Along the developmental process, several individuals from each instar were collected and preserved in 70% ethanol and then processed on the slides. We recorded the duration of development for each instar and described its larval features for the first time. The third instar larvae of H. fumipennis showed accessory oral sclerite present, anterior spiracle with 13-15 papillae, intersegmental spines mostly unicuspid with pointed end, and posterior spiracles heavily sclerotized with inter-slit projections. Some larval differences between H. fumipennis and Hypopygiopsis violacea were noted.
Analysis on fly artifacts produced by forensically important blowfly, Chrysomya megacephala (Fabricius) (Diptera:Calliphoridae), revealed several unique patterns. They can be divided into fecal spots, regurgitation spots and swiping stains. The characteristics of fecal spots are round with three distinct levels of pigmentation; creamy, brownish and darkly pigmented. Matrix of the spots appears cloudy. The round spots are symmetrical and non-symmetrical, delineated by irregular and darker perimeter which only visible in fairly colored fecal spots. Diameter of these artifacts ranged from 0.5 mm to 4 mm. Vomit or regurgitation spots are determined by the presence of craters due to sucking activity of blowflies and surrounded by thickly raised and darker colored perimeter. The size of these specks ranged from 1 mm to 2 mm. Matrix of the spots displays irregular surface and reflective under auxiliary microscope light. Swiping stains due to defecation by flies consists of two distinguishable segments, the body and tail. It can be seen as a tear drop-like, sperm-like, snake-like and irregular tadpole-like stain. The direction of body and tail is inconsistent and length ranged between 4.8 mm to 9.2 mm. A finding that should be highlighted in this observation is the presence of crater on tadpole-like swiping stain which is apparent by its raised border characteristic and reflective under auxiliary microscope light. The directionality of this darkly brown stain is random. This unique mix of regurgitation and swiping stain has never been reported before. Highlighting the features of artifacts produced by flies would hopefully add our understanding in differentiating them from blood spatters produced from victims at crime scenes.
Insects found associated with corpse can be used as one of the indicators in estimating postmortem interval (PMI). The objective of this study was to compare the stages of decomposition and faunal succession between a partially burnt pig (Sus scrofa Linnaeus) and natural pig (as control). The burning simulated a real crime whereby the victim was burnt by murderer. Two young pigs weighed approximately 10 kg were used in this study. Both pigs died from pneumonia and immediately placed in an oil palm plantation near a pig farm in Tanjung Sepat, Selangor, Malaysia. One pig was partially burnt by 1-liter petrol while the other served as control. Both carcasses were visited twice per day for the first week and once thereafter. Adult flies and larvae on the carcasses were collected and later processed in a forensic entomology laboratory. Results showed that there was no significant difference between the rate of decomposition and sequence of faunal succession on both pig carcasses. Both carcasses were completely decomposed to remain stage after nine days. The species of flies visiting the pig carcasses consisted of blow flies (Chrysomya megacephala, Chrysomya rufifacies, Hemipyrellia ligurriens), flesh fly (Sarcophagidae.), muscid fly (Ophyra spinigera), soldier fly (Hermetia illucens), coffin fly (Phoridae) and scavenger fly (Sepsidae). The only difference noted was in the number of adult flies, whereby more flies were seen in the control carcass. Faunal succession on both pig carcasses was in the following sequence: Calliphoridae, Sarcophagidae, Muscidae, Phoridae and lastly Stratiomyidae. However, there was overlap in the appearance of members of these families. Blowflies continued to oviposit on both carcasses. Hence postmortem interval (PMI) can still be estimated from the partially burnt pig carcass.
It is widely believed that most orchid flowers attract insects by using deception or chemical rewards in the form of nectar. Flowers of Bulbophyllum vinaceum produce a large array of phenylpropanoids that lure tephritid fruit fly males and also act as floral reward, which the flies subsequently convert to pheromone components. The major floral volatile components identified are methyl eugenol (ME), trans-coniferyl alcohol (CF), 2-allyl-4,5-dimethoxphenol (DMP), and trans-3,4-dimethoxycinnamyl acetate, whereas the minor components are eugenol, euasarone, trans-3,4-dimethoxy cinnamyl alcohol, and cis-coniferyl alcohol. Among the various floral parts, the lip (which is held in a closed position up against the sexual organs) has the highest concentration of the major compounds. An attracted male fly normally lands on one of the petals before climbing up onto and forcing the "spring loaded" floral lip into the open position, hence exposing the floral sexual organs. The architecture and location of chemical attractants of the lip compel the fly to align itself along the lip's longitudinal axis in a precise manner. As the fly laps up the compounds and moves towards the base of the lip, it passes the point of imbalance causing the lip to spring back to its normal closed position. The fly is catapulted headfirst into the column cavity, and its dorsum strikes the protruding sticky base of the hamulus and adheres to it. The momentum of the fly and the structural morphology of the long stiff hamulus act to pry out the pollinia from its anther cover. Hence, the pollinarium (pollinia + hamulus) is detached from the flower and adhered to the fly's dorsum. In this unique mutualistic association, both species receive direct reproductive benefits--the flower's pollinarium is transported for cross pollination, and the fly is offered a bouquet of phenylpropanoids (synomone) that it consumes, converts, and/or sequesters as sex pheromonal components, thus enhancing sexual attraction and mating success.
A body of an unknown adult female was found within a shallow burial ground in Malaysia whereas the skull was exposed and visible on the ground. During autopsy examination, nine insect larvae were recovered from the interior of the human skull and subsequently preserved in 70% ethanol. The larvae were greyish in appearance, each with a posterior elongated breathing tube. A week after the autopsy, more larvae were collected at the burial site, and some of them were reared into adults. Adult specimens and larvae from the skull and from the burial site were sequenced to obtain DNA barcodes. Results showed all adult flies reared from the burial site, as well as the larvae collected from the skull were identified as Eristalinus arvorum (Fabricius, 1787) (Diptera: Syrphidae). Here, we report the colonization of E. arvorum larvae on a human corpse for the first time.
In Malaysia, maggot debridement therapy (MDT) utilizes maggots of Lucilia cuprina (Wiedemann) to debride necrotic tissue from wound surface, reduce bacterial infection and therefore, enhance wound healing process. To evaluate the sterility of the sterile maggots produced after sterilization process before delivering onto patient wounds. Sterility of sterile maggots is crucial in ensuring the safe usage of MDT and patient's health. Eggs of L. cuprina collected from a laboratory colony were divided into treated group (sterilized) and control group (non-sterilized). Treated group underwent sterilization while eggs from control group were allowed to hatch without sterilization. Sodium hypochlorite and formaldehyde were the main disinfectants used in this sterilization process. Scanning electron microscope (SEM) was used to examine and ascertain the sterility of sterile maggots. SEM results showed that all sterilized L. cuprina eggs and maggots achieved sterility and all were cleared from bacterial contamination. In contrast, all non-sterilized eggs and maggots were found to be colonized by microorganisms. Sterilization method employed to sterilize eggs and maggots used in Malaysia MDT was proven successful and MDT is safe to be used as wound management tools.
Piophila casei (Linnaeus) (Diptera: Piophilidae) is reported from human cadavers in two separate forensic cases for the first time in Malaysia. Both bodies were found indoors. The first case, was that of a male of unknown nationality and age and also contained maggots of the muscid Ophyra spinigera (Stein). The second case was a female Chinese whose body also contained other species of maggots but these were not identifiable.
Misotermes mindeni Disney and Neoh is a solitary endoparasitoid of Macrotermes gilvus (Hagen) (Termitidae: Macrotermitinae), exclusively parasitizing the head of major soldiers. In this study, behavioral and morphological changes in the parasitized termite host were evaluated. We also observed the larval parasitoid emerging from the host. We found that parasitism mainly occurred in termite mounds overgrown with grass and mounds that had been broken up previously for other experiments. The parasitized soldiers showed a significantly lower level of interspecific aggressiveness compared with healthy soldiers (P < 0.05). Parasitized soldiers also changed in habitat preference to one isolated chamber of the nest. This might be an adaptive strategy that facilitates parasitoid dispersal, provides protection to parasitoids, and reduces the risk of parasitism to host colony. An abnormally rounded head capsule and remarkably short mandibles are characteristics of a parasitized soldier. The older larval fly stages were found only in major soldiers. We suggest that parasitization may first start in fourth or even earlier larval termite instars. The fly larva develops in the termite soldier's head capsule and pupates inside the host's body.
The majority of true parasitoids manipulate their host's physiology for their own benefit. In this study, we documented the physiological changes that occurred in major soldiers of the subterranean termite Macrotermes gilvus (Hagen) (Isoptera: Termitidae) parasitized by the koinobiont larval endoparasitoid Misotermes mindeni Disney and Neoh (Diptera: Phoridae). We compared the metabolic rate, body water content, body water loss rate, cuticular permeability, and desiccation tolerance between parasitized and unparasitized major soldiers. The metabolic rate of parasitized hosts was significantly higher than that of unparasitized termites. Mean total body water content of parasitized major soldiers (64.73±3.26%) was significantly lower than that of unparasitized termites (71.99±2.23%). Parasitized hosts also had significantly lower total body water loss rates (5.72±0.06%/h) and higher cuticular permeability (49.37±11.26 μg/cm/h/mmHg) than unparasitized major soldiers (6.75±0.16%/h and 60.76±24.98 μg/cm/h/mmHg, respectively). Parasitized major soldiers survived almost twice as long as unparasitized termites (LT(50)=6.66 h and LT(50)=3.40 h, respectively) and they had significantly higher tolerance to water loss compared to unparasitized termites (45.28±6.79% and 32.84±7.69%, respectively). Body lipid content in parasitized hosts (19.84±6.27%) was significantly higher than that of unparasitized termites (6.17±7.87%). Finally, parasitized hosts had a significantly lower percentage of cuticular water content than unparasitized major soldiers (10.97±1.84% and 13.17±2.21%, respectively). Based on these data, we conclude that the parasitism-induced physiological changes in the host are beneficial to the parasitoids as the alterations can clearly increase the parasite's chances of survival when exposed to extreme environmental conditions and ensure that the parasitoids are able to complete their larval development successfully before the host dies.
The larval parasitoid Verticia fasciventris Malloch (Diptera: Calliphoridae) develops in the head of soldiers of the fungus-growing termite Macrotermes carbonarius (Hagen) (Isoptera: Termitidae). Morphological and behavioral changes in the host were evaluated and the termite castes and stages that were parasitized were identified. The larval emergence process is also described and possible mechanisms for the parasitoid fly's entry into the host body are discussed based on qualitative observations. Only a single larva per host was found. The mature larva pupated outside the host's body by exiting between the abdominal cerci. Parasitized soldiers possess a short and square-shaped head capsule, a pair of notably short mandibles, and a pair of 18-segmented antennae. Although parasitized soldiers were statistically less aggressive than healthy soldiers (P < 0.05), they expressed varying levels of aggression. Both minor and major soldiers can be parasitized and based on evidence from presoldiers, parasitization may begin during the precursor stages of soldiers. However, the stage at which parasitism first occurs has not been determined.