DNA identification of blow fly species can be a very useful tool in forensic entomology. One of the potential benefits that mitochondrial DNA (mtDNA) has offered in the field of forensic entomology is species determination. Conventional identification methods have limitations for sibling and closely related species of blow fly and stage and quality of the specimen used. This could be overcome by DNA-based identification methods using mitochondrial DNA which does not demand intact or undamaged specimens. Mitochondrial DNA is usually isolated from whole blow fly and legs. Alternate sources for mitochondrial DNA isolation namely, egg, larva, puparium and empty puparium were explored in this study. The sequence of DNA obtained for each sample for every life cycle stage was 100% identical for a particular species, indicating that the egg, 1st instar, 2nd instar, 3rd instar, pupa, empty puparium and adult from the same species and obtained from same generation will exhibit similar DNA sequences. The present study also highlighted the usefulness of collecting all life cycle stages of blow fly during crime scene investigation with proper preservation and subsequent molecular analysis. Molecular identification provides a strong basis for species identification and will prove an invaluable contribution to forensic entomology as an investigative tool in Malaysia.
Estimation of post-mortem interval (PMI) is crucial for time of death determination. The advent of DNA-based identification techniques forensic entomology saw the beginning of a proliferation of molecular studies into forensically important Calliphoridae (Diptera). The use of DNA to characterise morphologically indistinguishable immature calliphorids was recognised as a valuable molecular tool with enormous practical utility. The local entomofauna in most cases is important for the examination of entomological evidences. The survey of the local entomofauna has become a fundamental first step in forensic entomological studies, because different geographical distributions, seasonal and environmental factors may influence the decomposition process and the occurrence of different insect species on corpses. In this study, calliphorids were collected from 13 human corpses recovered from indoors, outdoors and aquatic conditions during the post-mortem examination by pathologists from the government hospitals in Malaysia. Only two species, Chrysomya megacephala and Chrysomya rufifacies were recovered from human corpses. DNA sequencing was performed to study the mitochondrial encoded COI gene and to evaluate the suitability of the 1300 base pairs of COI fragments for identification of blow fly species collected from real crime scene. The COI gene from blow fly specimens were sequenced and deposited in GenBank to expand local databases. The sequenced COI gene was useful in identifying calliphorids retrieved from human corpses.
Epilepsy is a common neurological disorder characterized by seizures which result in distinctive neurobiological and behavioral impairments. Not much is known about the causes of epilepsy, making it difficult to devise an effective cure for epilepsy. Moreover, clinical studies involving epileptogenesis and ictogenesis cannot be conducted in humans due to ethical reasons. As a result, animal models play a crucial role in the replication of epileptic seizures. In recent years, non-mammalian models have been given a primary focus in epilepsy research due to their advantages. This systematic review aims to summarize the importance of non-mammalian models in epilepsy research, such as in the screening of anti-convulsive compounds. The reason for this review is to integrate currently available information on the use and importance of non-mammalian models in epilepsy testing to aid in the planning of future studies as well as to provide an overview of the current state of this field. A PRISMA model was utilized and PubMed, Springer, ScienceDirect and SCOPUS were searched for articles published between January 2007 and November 2017. Fifty-one articles were finalized based on the inclusion/exclusion criteria and were discussed in this review. The results of this review demonstrated the current use of non-mammalian models in epilepsy research and reaffirmed their potential to supplement the typical rodent models of epilepsy in future research into both epileptogenesis and the treatment of epilepsy. This review also revealed a preference for zebrafish and fruit flies in lieu of other non-mammalian models, which is a shortcoming that should be corrected in future studies due to the great potential of these underutilized animal models.
Larvae of a gall midge were found feeding on the fungal rust Maravalia pterocarpi (Pucciniomycetes: Pucciniales: Chaconiaceae) infesting the economically important sua tree Dalbergia tonkinensis (Fabaceae) on Hainan Island, China. The adults, pupae and larvae were collected, their morphology was studied and a segment of the Cytochrome Oxidase unit I (COI) mitochondrial gene was sequenced. The gall midge proved to be a species new to science that belongs to the genus Mycodiplosis (Diptera: Cecdiomyiidae). Comparison of the sequence to published Cecdiomyiidae sequences revealed that, despite being undescribed and unnamed, it was previously found in east and south-east Asia to feed on several rust species: Puccinia coronata (Pucciniomycetes: Pucciniales: Pucciniaceae) that develops on Lolium multiflorum (Poaceae), Puccinia sp. on Zea mays (Poaceae), Puccinia arachidis on Arachis hypogaea (Fabaceae) and Puccinia allii on Allium fistulosum (Amaryllidaceae). The new species is described and named here Mycodiplosis puccinivora Jiao, Bu Kolesik. It occurs in China, Japan, Thailand, Bangladesh and possibly Malaysia and Australia. In Hainan it has four to five generations per year.
Meliponiculture, the keeping of domesticated stingless bees such as Geniotrigona thoracica (Smith, 1857) (Hymenoptera: Apidae), is an increasingly popular agricultural industry in Malaysia. This study reports the soldier fly (Diptera: Stratiomyidae) species of the genus Hermetia colonizing stingless bee colonies in Malaysia. The larvae were reared in the laboratory to the adult stage and identified through molecular and morphological approaches. Hermetia illucens (Linnaeus, 1758) and Hermetia fenestrata de Meijere, 1904 (Diptera: Stratiomyidae) were identified from the sample provided. Earlier records of stratiomyids in stingless bee nests were misidentified as H. illucens. This paper represents the first identified record of H. fenestrata colonizing a "spoiled" stingless bee colony. In addition, adult and larval morphological differences between both species and the roles of both species in bee nest decomposition are discussed.
The larval development of Chrysomya albiceps and Lucilia sericata is a well-known and valuable tool for estimating the postmortem interval (PMI). The third instar larvae of Ch. albiceps are facultative predators of the larvae of other necrophagous dipterans, and this behaviour is undoubtedly an important factor to consider. Both species are found together during human autopsies in Spain and other parts of Europe, where they cohabit, so a precise knowledge of their larval coexistence data is essential. The aim of this study is to investigate their coexistence and the intraguild predation of Ch. albiceps on L. sericata under experimental conditions and in real case reports. To analyse intra- and interspecific competition, four densities were used under controlled abiotic conditions [25ºC, 60-70% RH and 12:12 (D:N)]. The experimental data were compared with data corresponding to their coexistence in natural conditions, so annual activity and forensic case reports for both species were studied. The results indicate that the mortality of immature specimens in both species, the preimaginal developmental time, and adult size were affected by competition. In natural conditions, adult coexistence during the spring-summer period was confirmed. The simultaneity of the two species colonising human corpses is frequent in the studied area, with Ch. albiceps being the dominant species, and L. sericata the relevant species for estimating the minimum postmortem interval.
This case study reports on two unrecorded Coleopteran species found together on a human corpse in Malaysia. The mummified human remains were discovered in a house in Selangor, Malaysia. The pathologist confirmed that the death was due to a traumatic chest injury. Maggots, beetles, and fly pupal casings were found mainly on the front part of the body. Empty puparia of Diptera were collected during the autopsy and identified later as the muscid Synthesiomyia nudiseta (van der Wulp, 1883) (Diptera: Muscidae). Also, the insect evidence received included larvae and pupae of Megaselia sp. (Diptera: Phoridae). According to the insect development data, the minimum postmortem period was estimated by the time to reach the pupal developmental stage (in days). The entomological evidence included the first record of Dermestes maculatus De Geer, 1774 (Coleoptera: Dermestidae) and Necrobia rufipes (Fabricius, 1781) (Coleoptera: Cleridae), which have not previously been recorded on human remains in Malaysia.
The estimation of the postmortem interval (PMI) is an essential information in death investigations. It is necessary to know the developmental data of the most important necrophagous insect species in every geographical area. Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) is one of the most common species associated with human body decomposition, especially in the tropics, so a precise knowledge of its life cycle is crucial. However, despite its ubiquity, developmental data in a range of tropical temperatures is scarce. For this reason, the aim of this study is to provide the developmental data of the blowfly, C. megacephala, in a range of tropical temperatures and to apply these data in forensic entomology. Four temperatures were examined (i.e., 27.0°C 29.5°C, 32.0°C and 34.5°C) and the time of developments from egg to adult were recorded. To build the growth curves, five larvae per day were measured with a digital caliper. Accumulated degree-days were calculated and the isomorphen diagram for this species was plotted. As we predicted, the results showed that the life cycle of this species was affected by the increasing temperature. The implications of these new data for determining the minimum PMI in forensic entomology were discussed.
A partially skeletonized human corpse was found in bushes in Selangor, Malaysia in June 2020. Entomological evidence was collected during the autopsy and sent to the Department of Medical Microbiology and Parasitology, Faculty of Medicine, Universiti Teknologi MARA (UiTM) for minimum postmortem interval (PMImin) analysis. Standard protocols were applied when processing preserved and live insect specimens of both larval and pupal stages. Entomological evidence revealed that the corpse was colonized by Chrysomya nigripes Aubertin, 1932 (Diptera: Calliphoridae) and Diamesus osculans (Vigors, 1825) (Coleoptera: Silphidae). Chrysomya nigripes was chosen as the PMImin indicator as this fly species is an earlier colonizer compared to D. osculans beetle larvae which their presence is the indicative of late stage of decomposition. For the present case, the pupae of C. nigripes were the oldest insect evidence collected and based on the available developmental data, the estimated minimum PMI was established between 9 and 12 days. It is noteworthy to highlight that this is the first record of D. osculans colonization on a human corpse.
A key to the common Calliphorid flies of Peninsular Malaysia is presented. Illustrations of the genitalia of some rare species, list of new localities, altitudes and other ecological data are also presented. The following species were recorded for the first time from Peninsular Malaysia; Catapicephala sinica, C. kurahashii, Taninanina javanica, Hemipyrellia tagaliana, Lucilia sinensis, Blaesoxipha kasterni, Boettcherisca javanica, Parasarcophaga misera, P. orchidea, P. albicephs, Sarcosolomonia crinita, Thyrsocnema bornensis, Sarcorohdendorfia antilope and Lioproctia pattoni.
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.
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.
This study was carried out in Agricultural Park, Teluk Cempedak and Bukit Pelindung at Kuantan, Pahang in October 2007. These three areas were different in ecological characteristic, Agricultural Park is a lowland region in Kuantan rural area, Teluk Cempedak is Kuantan’s most famous beach, and Bukit Pelindung is a reserved rainforest which is 200 meters from the sea level. Fly specimens were collected using four different kinds of baits: dry prawn, salted fish, pork and mango. Each of these baits was placed in a plastic container and exposed for one hour to attract flies. Within 5 minutes, flies started swarming around the baits. The flies were more attracted to the pork and salted fish compared to the other two baits. Fifty one flies, one moth (Lepidoptera) and one wasp (Hymenoptera) were collected. In Agricultural Park, two Lucilia cuprina, one Chrysomya megacephala and one Sarcophaga sp. were collected. For Teluk Cempedak beach, there were two Sarcophagids, 31 Chrysomya megacephala, five Musca domestica, one Lucilia cuprina and one moth were caught. Flies collected from Bukit Pelindung included five C.megacephala, two Sarcophagids, one Musca domestica and one wasp. Most C.megacephala were attracted to the pork and salted fish.
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.
Under normal circumstances, insects such as blow flies will oviposit and larvae will colonize a carcass as soon as possible. However, insect colonization on a carcass may be delayed due to the effects of wrapping, shallow burial, addition of lime derivatives to mitigate scavenging and odor, or extreme weather. The impacts of delayed insect colonization on carcass decomposition and its subsequent effect on soil chemistry profiles have not been examined to date. The objectives of this study were to determine soil chemistry dynamics associated with porcine carcasses experiencing delayed insect colonization for 7-day or 14-day. Soil chemistry profiles such as ammonium-N (NH4 -N), orthophosphate-P (PO4 -P), and dissolved organic carbon (DOC) were significantly different among treatments: insect inclusion (immediate access of blow fly colonization on porcine carcasses), 7-day insect exclusion and 14-day insect exclusion (blow fly access was delayed up to 7-day and 14-day). Furthermore, significant differences of soil chemical profiles were detected between days of decomposition and soil regions. Soil moisture, NH4 -N, PO4 -P, and DOC were significantly higher when insects were excluded from the porcine carcass suggesting loss of tissue from larval feeding reduced the mass of nutrients entering the soil. This study provides useful information for forensic science in cases where insect colonization is delayed for a period of time postmortem and soil chemistry in the cadaver decomposition island is considered for estimating postmortem interval.
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.
Isomyia paurogonita Fang & Fan, 1986 (Diptera: Calliphoridae), a rare species of the subfamily Rhiniinae (tribe Cosminini) was recorded for the first time in Malaysia. We collected one male and two females during a field trip conducted at Genting Highland, Pahang, peninsular Malaysia in May 2011. A 3-day old cow liver was offered as attractant and dipterans collected were transferred to the laboratory for specimens processing and identification. The adults of I. paurogonita were attracted to the odour and then captured by using a sweep net. Isomyia paurogonita was also recorded from two other localities in Peninsular and Malaysian Borneo, namely Gombak Utara, Selangor and Sibu, Sarawak.
This preliminary study was carried out in a palm oil plantation in Tanjung Sepat, Selangor in 17 May 2007 by using pig (Sus scrofa) as a carcass model in forensic entomological research. A 3 month old pig (8.5 kg) that died of pneumonio was placed in the field to observe the decomposition stages and the fauna succession of forensically important flies. Observation was made for two weeks; two visits per day and all climatological data were recorded. The first visitor to the pig carcass was a muscid fly, seen within a minute, and followed by ants and spiders. Within half an hour, calliphorid flies came over. On the second day (fresh), few calliphorid and sarcophagid flies were found on the carcass. Two different species of moths were trapped in the hanging net. The first larva mass occurred on the third day (bloated) around the mouthpart, with some L1 and L2 found in the eyes. Reduvid bugs and Staphylinidae beetles were recovered on the fourth day (active decay), and new maggot masses occurred in the eyes and anus. L3 larvae could be found beneath the pig carcass on the fourth day. On the fifth day (active decay), new maggot masses were found on neck, thorax, and hind legs. Advance decay occurred on the sixth day with abundant maggots covering all over the body. The main adult fly population was Chrysomya megacephala (day 2 to day 6), but the larvae population was mainly those of Chrysomya rufifacies (day 4 to day 14). The dry stage began on the eighth day. Hermetia illucens adult was caught on day-13, and a larvae mass of Chrysomya rufifacies was seen burrowing under the soil. This forensic entomological research using pig carcass model was the first record in this country.
During a forensic entomological study conducted at an oil palm plantation in Tanjung Sepat, Kuala Langat, Selangor, a Bengalia emarginata Malloch, 1927 (Diptera: Calliphoridae: Calliphorinae: Bengalini) was collected for the first time. Two adults were collected nearby the pig carcass by the first author and identified by the second. Prior to this finding, nine species of Bengalia were recorded from peninsular Malaysia or Borneo. Male of B. emarginata are different from Bengalia varicolor Fabricious by the following characters: Sternite 5 projection rounded with small identation and mid tibia double-fringed in ventral surface.