New developmental data of Chrysomya megacephala (Diptera: Calliphoridae) in tropical temperatures and its implications in forensic entomology

@#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.

The first record of parasitic mite, Leptus sp. (Acari: Erythraeidae) associated with a necrophagous fly, Chrysomya villeneuvi Patton, 1922 (Diptera: Calliphoridae)

@#This is the first documentation of parasitic mite, Leptus sp., found on a necrophagous blowfly, Chrysomya villeneuvi collected from a decomposing wild boar carcass placed in Taman Negara (National Park), Kuala Keniam, Pahang, Malaysia. Blowflies around the carcass were captured using an insect net before being examined under a stereomicroscope. Upon microscopic observation, we found a mite attached on the scutellum of C. villeneuvi adult. The mite was carefully removed and preserved in 70% ethanol subsequently. Then, the mite was cleared in lactophenol before being mounted in Hoyer’s medium. The morphological identification of the mite was conducted and Leptus sp. was identified. The species belongs to the phalangii species group and the aldonae species subgroup. This study highlights the new association of Leptus sp. and C. villeneuvi for the first time.

Juvenile hormone: Production, regulation, current application in vector control and its future applications

@#Juvenile hormone is an exclusive hormone found in insects which involves regulating various insect physiology. A total of eight juvenile hormones have been identified in insects which include JH 0, JH I, JH II, JH III, 4-methyl JH I (Iso- JH 0), JHB III, JHSB III, and MF. Corpora allata are the glands responsible for the production and synthesis of these hormones. They are involved in moulting, reproduction, polyethism, and behavioural regulations in different orders of insects. Factors such as diet temperatures, photoperiods, and plant compounds affect the biosynthesis and regulation of juvenile hormones. Juvenile hormones analogue is usually used to disrupt normal regulation of JH and this analogue is categorized as insect-growth regulators (IGRs) and is widely used in pest control as an alternative to chemical insecticides. Other applications of biosynthesis activities of this hormone have not been explored in the area of JHs. In this review, current applications of JHs with an addition of their future application will be discussed.

Preliminary observation on the lepidopteran colonization on rat and rabbit carcasses in Malaysia

@#The immature stages of necrophagous insects such as Diptera and Coleoptera play a vital ecological role in carrion decomposition. These invertebrates reduce the necromass significantly through consumption and recycle nutrients into organic forms which are readily being used by autotrophs or served as an abiotic storage in the soil ecosystem. Fly and beetle larvae are frequently encountered decomposers on ephemeral resource patches; however, lepidopterans associated with carrion decomposition is seldom reported. Here, we report colonization of Monopis sp. (Tineidae) and an unknown species of Psychidae on a rat carcass, and a Lithosiini caterpillar (Arctiidae) on a rabbit carcass in Peninsular Malaysia for the first time. The feeding behaviour and their potential forensic implications are discussed.

The First Report of Lasioderma Serricorne Infestation on Dried Fish Crackers in Malaysia (Liputan Pertama Infestasi Lasioderma Serricorne pada Keropok Ikan di Malaysia) / Jurnal Sains Kesihatan Malaysia

@#This paper is the first record of cigarette beetles collected from dried fish crackers (also known as “keropok ikan” in Malay) in Malaysia. The dead cigarette beetles were firstly isolated from a packet of dried fish crackers and were subsequently kept in 70% ethanol. The beetles were then identified as Lasioderma serricorne (Fabricius 1792) (Coleoptera: Anobiidae). They are common pests of stored products such as tobacco, flour, and cocoa beans but there is no record of this beetle infestation on dried fish crackers in Malaysia.

Age estimation of forensically important blowfly, Chrysomya megacephala (Diptera: Calliphoridae) pupae using micro-computed tomography imaging

@#Accurate estimation of the minimum post-mortem interval (minPMI) is important in the investigation of forensic cases. Various thanatological methods are being used to estimate this interval. However, entomology approach is the most reliable method for this minPMI estimation especially when death has occurred over 72 hours and involved insects or other arthropods evidence at the death scene. The current methods of age estimation are daunting and destructive especially when dealing with pupal stage. The aims of this study were to characterize the morphological changes during intra-puparial period of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) and their relation with minPMI estimation by using a high resolution micro-Computed Tomography (micro-CT). Gravid C. megacephala were collected from a rural area in Sungai Buloh, Selangor and cultured in the laboratory at 23.83±0.25°C with light: dark hour of 12:12 to initiate oviposition. The resulting larvae were reared until pupal stage. A pupa was collected at first (24 hours), second (48 hours), third (72 hours), and fourth quarter (96 hours) of the intra-puparial period. The pupal samples were placed directly into 70% ethanol for preservation. Micro-CT scanning was employed to acquire microstructural information following pupal sample staining for contrast enhancement. Eight age-informative internal morphological landmarks were mapped from the micro-CT scanning. The present study enhanced the potential value of micro-CT for the estimation of minPMI based on the internal morphological changes of C. megacephala pupae. This novel method is a promising tool for improving medico-legal investigations in forensic entomology.

Detection of human Sarcocystosis using dried blood on filter papers: An Immunofluorescent Antibody Test

@#Sarcocystosis, a parasitic infection caused by a protozoa belonging to the genus Sarcocystis, is found worldwide in both and animals. Sarcocystis spp., require two animal hosts to complete their life cycle. The infection has gathered more global attention after recent outbreaks, especially amongst wester travellers to Malaysia. Other than sporadic cases and the current outbreaks, little information is available regarding human Sarcocystis infection in Malaysia. The present study aims to determine the prevalence of sarcocystosis among humans using an immunofluorescent antibody (IFA) test applied to dried blood on filter papers. A total of 200 blood samples were collected on filter papers from autopsy cases at two Malaysian hospitals: Sungai Buloh Hospital (peninsular Malaysia) and Queen Elizabeth Hospital (Malaysian Borneo). Antigens were prepared from bradyzoites harvested from positive goats’ muscle samples. Of the 200 samples, 32 (16%) had Sarcocystis antibodies that showed positive fluorescence reactions on filter papers. There was no significant difference (t-test, p value > 0.05) in prevalence rates between samples collected from autopsies at peninsular Malaysia and Malaysian Borneo. The results demonstrated that the filter paper technique can be used as one of the alternative serological tests in the diagnostic of human sarcocystosis.

First report of Histiostoma sp. (Astigmata: Histiostomatidae) phoretic on a forensically important blowfly, Chrysomya villeneuvi (Diptera: Calliphoridae)

@#This is the first record of phoretic histiostomatid mites found on a forensically important blow fly species, Chrysomya villeneuvi (Diptera: Calliphoridae), collected from decomposing rabbit carcasses placed in Bukit Lagong Forest Reserve, Sungai Buloh and MARDI Cameron Highlands, Malaysia. The blow flies frequenting around the carcasses were first captured using an insect net. After pinning, they were examined under a stereomicroscope and mites phoretic on their body were carefully removed and preserved in 70% ethanol. Mites were cleared in lactic acid before mounting on slides using Hoyer’s medium and identified under a compound microscope. The flies and their mites were identified as C. villeneuvi and deutonymphs of Histiostoma spp. (Astigmata: Histiostomatidae), respectively. This insectmite association may be useful to provide insights regarding the minimum post-mortem interval and the location of death in forensic entomological investigations.

Light microscopy and molecular identification of Sarcocystis spp. in meat producing animals in Selangor, Malaysia

One thousand and forty-five tissue samples of skeletal muscles, tongue, heart, diaphragm and esophagus were collected from 209 animals (43 sheep, 89 goats and 77 cattle) from an abattoir in Selangor between February and October, 2013. Each sample was divided into three pieces with each piece measuring 2-3 mm3. Each piece was then squeezed between two glass slides and examined microscopically at x 10 magnification for the presence of sarcocystosis. Three positive samples from each animal species were then fixed in 10% formalin for histological processing. Seven positive samples collected from each animal species were preserved at -80°C or 90% ethanol for gene expression studies. Microsarcocysts were detected in 114 (54.5%) animals by light microscopy (LM). The infection rates in sheep, goat and cattle were 86, 61.8 and 28.6% respectively. The highest rate of infection was in the skeletal muscles of sheep (64.9%) and goats (63.6%) and in the heart of cattle (63.6%). The cysts were spindle to oval in shape and two stages were recognized, the peripheral metrocytes and centrally located banana-shaped bradyzoites. 18S rRNA gene expression studies confirmed the isolates from the sheep as S. ovicanis, goats as S. capracanis and cattle as S. bovicanis. This, to the best of our knowledge, is the first molecular identification of an isolate of S. ovicanis and S. capracanis in Malaysia. Further studies with electron microscopy (EM) are required in the future to compare the features of different types of Sarcocysts spp.

Morphological descriptions on the larvae of Hypopygiopsis fumipennis (Walker, 1856) (Diptera: Calliphoridae)

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.

New locality record of Isomyia paurogonita Fang & Fan, 1986 (Diptera: Calliphoridae) from Peninsular Malaysia and Borneo

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.

A new record for Lispe orientalis Wiedemann, 1824 (Diptera: Muscidae) from peninsular Malaysia

Lispe orientalis Wiedemann, 1824 is recorded for the first time in peninsular Malaysia. Specimens were collected from a mushroom cultivation farm in Genting Highlands, Pahang (3°25’18"N 101°47’48"E). Previously, this species had been recorded from Azerbaijin, India, Russia, Tajikistan, Thailand, Turkey and South Korea. The male of Lispe orientalis can be determined by the following characteristics: body non-metallic, ashy gray, third antennal segment black, R5 cell not narrow apically, hind metatarsus normal, legs entirely black, femora with long bristle-like hairs on av and pv surfaces, hind tibia without av and pv seta and the palpi orangish in colour.