First observation of Afromorgus chinensis (boheman, 1858) (Coleoptera: Trogidae) on a rabbit (Oryctolagus cuniculus L., 1758) carcass and its implications in forensic entomology.

Beetles (Coleoptera) are known to constitute forensic evidence in medico-legal investigations as their presence can be used to date human remains in almost all decomposition stages. Many forensic studies focus on the successional colonization pattern of flies (Diptera); however, beetles have not so far been studied extensively for this aspect. A beetle of the genus Afromorgus Scholtz, 1986, A. chinensis (Boheman, 1858) (Scarabaeoidea: Trogidae), was found beneath a late decaying rabbit carcass at Paya Indah Wetland, Dengkil, Malaysia, for the first time. Both genus and species are already known to occur in Malaysia from literature.

A new host record of Haemaphysalis semermis (Acari: Ixodidae) from the domestic cat (Felis catus) (Carnivora: Felidae) in Orang Asli village, Pahang National Park Malaysia.

We report the presence of a male Haemaphysalis semermis collected from the domestic cat, Felis catus in an aboriginal village located in Pahang, Malaysia. This paper constitutes a new host record of this tick species, and also the first documentation of the infestation of companion animals other than domestic dogs (Canis lupus) by H. semermis in Malaysia. Additionally, we have included an updated host index of the tick species in Southeast Asia.

Two new records of chewing lice (Phthiraptera: Amblycera) from the Oriental honey buzzard [Pernis ptilorhynchus (Temminck, 1821)] and house crow (Corvus splendens Viellot, 1817) in Malaysia.

We report two new records of chewing lice from avian pets in Peninsular Malaysia: Colpocephalum apivorus Tendeiro, 1958 from an Oriental honey buzzard (Pernis ptilorhynchus (Temminck, 1821)), and Myrsidea splendenticola Klockenhoff, 1973 from an albino house crow (Corvus splendens Vieillot, 1817). The scarcity of louse records from avian pets and wild birds, and the lack of louse research in Malaysia are discussed.

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.

Rhipicephalus annulatus, R. australis or R. microplus? Discordance between morphological and genetic data among three cattle tick species.

The taxonomy of ticks of the subgenus Boophilus has been extensively debated and is often complicated by the high intraspecific variation of morphological features between species. Notably, the cattle tick Rhipicephalus microplus is a species complex consisting of Rhipicephalus annulatus, Rhipicephalus australis and the three mitochondrial clades (A-C) of R. microplus. To gain insight into the taxonomic status of this species complex, we performed morphological and molecular analyses on these cattle ticks across four states in peninsular Malaysia. We morphologically identified 60 males and 104 females of R. microplus, 298 males and 374 females of R. australis, and one R. annulatus male in our field collection, of which the latter two species have never been recorded in Malaysia. However, all three morphologically identified species were molecularly assigned as R. microplus clade A based on the barcoding cytochrome oxidase subunit I (COI) analysis. The discrepancy between morphological and genetic data highlights an urgent need for further exploration and in-depth research into the taxonomic status of these sympatric tick species.

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.

Acari community in association with delayed pig carrion decomposition.

Acari community structure and function associated with delayed pig carrion decomposition has not been examined. In this study, 18 swine carcasses were studied in central Texas, USA, during two consecutive summers (2013 and 2014). Samples of ca. 400 g soil were collected from beneath, aside, and 5 m away from each pig carcass over 180 days. Mites from soil samples were extracted using Berlese funnels and identified to order and family levels and classified according to ecological function. In total 1565 and 1740 mites were identified from the 2013 and 2014 soil samples, respectively. Significant differences were determined for mite community structure at order and family levels temporally on carrion (e.g., day 0 × day 14) regardless of treatments and between soil regions where mites were collected (e.g., soil beneath vs. soil 5 m away from carrion). However, no significant differences were found in mite community structure at the order level between pig carrion with and without delayed Diptera colonization (i.e., treatments). Analysis at the family level determined a significant difference across treatments for both summers. Ecological function of mites did not change significantly following the delayed decomposition of pig carcasses. However, detritivores and fungivores were significant indicator groups during the pig carrion decomposition process. Furthermore, 13 phoretic mite species associated with eight forensically important beetle species were documented. Data from this study indicated that the rate of nutrient flow into the soil impacted associated arthropod communities; however, detecting such shifts depends on the taxonomic resolution being applied.

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.

Soft ticks (Acari: Argasidae) in the island nations of Southeast Asia: A review on their distribution, associated hosts and potential pathogens.

Soft ticks (Acari: Argasidae) are the second major family of the blood feeding metastriates and vectors of a number of viral and bacterial pathogens for both humans and animals. Despite the growing effort on tick surveillance and studies worldwide, there is still limited information on the soft tick distribution in the island nations of Southeast Asia, especially species that are medically and veterinarily important. With the aim to provide an overview of the current status of knowledge on soft tick distribution in the island nations of Southeast Asia (Malaysia, Singapore, Brunei, Indonesia, the Philippines and Timor-Leste), this article reviews the species of soft ticks (Acari: Argasidae) and their associated hosts and pathogens, with the addition of a pictorial summary and list of tick species discovered in this region. The most prevalent soft tick genus is Carios, and the host species most associated with findings of soft ticks in this region are bats, particularly of the Pteropodidae and Vespertilionidae families. Furthermore, the only known pathogen originating from soft ticks in the island nations of Southeast Asia was the Keterah virus, which was isolated from Argas pusillus tick in Malaysia.

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.

Review of Mites Found on Various Animal Hosts and at Different Localities in Malaysia.

Mite biodiversity and distribution in Malaysia is currently understudied. Most previous works on Malaysian Acari have focused on pest organisms of medical, veterinary, and agricultural concern, with a few recent studies centered on mites in forensic contexts. Previous literatures have targeted collection sites in forest reserves and/or mountains in either Peninsular or Malaysian Borneo, though the state of Sarawak had the least publications related to mite species descriptions despite having the highest number of nature parks of any state in the country. Most publications focused on the three states Selangor, Pahang and Sabah. Most of the mite species reported were from mammals (66.3%), with fewer species from birds (21.7%), arthropods (11.2%), and reptiles (0.8%). We believe that further work on the systematic documentation of mite species throughout Malaysia is necessary as it could generate useful tools, such as the use of mites as biogeographical markers or as forensic indicators. Therefore, this review catalogs mite species that have been documented in or on animal hosts in Malaysia and serves as a foundation for future work.

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.

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.

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.

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.

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.

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.