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

Azmiera N, Mariana A, Heo CC

Trop Biomed, 2019 Dec 01;36(4):1099-1104.
PMID: 33597479

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

Azmiera N, Mariana A, Pimsler ML, Heo CC

J Med Entomol, 2020 09 07;57(5):1354-1363.
PMID: 32440683 DOI: 10.1093/jme/tjaa086

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.
Colonization of Rabbit Carcasses by Drain Fly Larvae, Psychoda sp. (Diptera: Psychodidae): The First Report

Azmiera N, Low VL, Heo CC

Acta Parasitol, 2021 Jun;66(2):706-709.
PMID: 33389626 DOI: 10.1007/s11686-020-00313-z

INTRODUCTION: Psychoda sp. is often collected from patchy habitats such as sewers, drains and decomposing organic matters. The discovery of Psychoda sp. in forensic studies indicated that it might have noteworthy value in assisting death investigations.
PURPOSE: This study reports on the first finding of Psychoda larvae collected from decomposing rabbit carcasses placed in Cameron Highlands, Pahang, Malaysia.
METHODS: The larvae were first observed on rabbit carcasses and were collected using tweezers and carefully preserved in 70% ethanol. They were subsequently mounted on microscopy slides using Hoyer's medium and identified as Psychoda sp. morphologically. The identification was also confirmed through a DNA barcoding analysis.
RESULTS: Psychoda sp. larvae were collected on day-10 post-mortem where the rabbit carcasses were at the advanced decay stage of decomposition. The cytochrome c oxidase I (COI) gene sequences of the larvae had 90% similarity with the Psychoda spp. in the database.
CONCLUSION: The finding of these larvae on carrion may provide additional valuable insights into forensic entomology and may assist in death investigations.
Fulltext Human and animal invasive muscular sarcocystosis in Malaysia--recent cases, review and hypotheses

Tappe D, Abdullah S, Heo CC, Kannan Kutty M, Latif B

Trop Biomed, 2013 Sep;30(3):355-66.
PMID: 24189667

Sarcocystosis, an unusual parasitic zoonotic disease, is caused by coccidian/ apicomplexan protozoa in humans and animals. The parasites usually develop in a heteroxenous predator-prey life-cycle involving final (carnivore) and intermediate (omnivore/herbivore) hosts. Besides the intestinal, non-invasive form of the disease in which humans and animals are the definitive hosts for certain Sarcocystis spp., the invasive form has come to recent attention. In the latter, humans and animals serve as intermediate host harbouring sarcocysts in their muscle tissue. Already in 1991 sarcocystosis was seen as a potential emerging food borne zoonosis in Malaysia, and in 2011 and 2012 the largest cluster of symptomatic human muscular sarcocystosis world-wide was reported from Tioman Island, Pahang state. In this review, we focus on invasive sarcocystosis in humans and animals in Malaysia, review the recorded cases and epidemiology, and present hypotheses.
Fulltext New locality record of Isomyia paurogonita Fang & Fan, 1986 (Diptera: Calliphoridae) from Peninsular Malaysia and Borneo

Heo CC, Aisha S, Kurahashi H, Omar B

Trop Biomed, 2013 Mar;30(1):159-63.
PMID: 23665723 MyJurnal

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.
Fulltext On the predation of fly, Chrysomya rufifacies (Macquart) by a spider, Oxyopes sp. Latreille (Oxyopidae)

Heo CC, Mohamad AM, John J, Baharudin O

Trop Biomed, 2008 Apr;25(1):93-5.
PMID: 18600210 MyJurnal

During a forensic entomological study conducted in a palm oil plantation in Tg.Sepat, Selangor in September 2007, a spider (Arachnida), Oxyopes sp. (Oxyopidae) was found to predate on a calliphorid fly (Chrysomya rufifacies). The female spider laid a silk thread, or "drag line", behind it as it moved. This spider bites its prey by using a pairs of chelicerae, and injecting venom into the fly. The fly was moving its wing trying to escape, however, it succumbed to the deadly bite.
Fulltext Insect succession on a decomposing piglet carcass placed in a man-made freshwater pond in Malaysia

Heo CC, Mohamad AM, John J, Baharudin O

Trop Biomed, 2008 Apr;25(1):23-9.
PMID: 18600201

This entomological study was conducted in a man-made freshwater pond in a palm oil plantation in Tanjung Sepat, Selangor from 23 July 2007 by using pig (Sus scrofa) as a carcass model. A 1.5 month old piglet (5 kg), which died of asphyxia after being accidentally crushed by its mother, was thrown into a pond. Observation was made for ten days; one visit per day and climatological data were recorded. On the first two days, the piglet carcass sunk to the bottom of the pond. The carcass floated to the surface on the third day but no fly activities were seen. The blow fly, Chrysomya megacephala and Chrysomya rufifacies started to oviposit on the fourth day. Other than adult flies, a spider (Arachnida) was also observed on the carcass. Bubbles accumulated at the mouthpart, and the abdomen was greenish black. A lot of blow fly eggs were seen on the body surface on the fifth day (floating decay), along with first and second instars C. megacephala crawling under the piglet's skin. On the sixth day, adult blow fly, C. megacephala,and C. rufifacies,and muscid flies, Ophyra spinigera and Musca domestica were observed on to the carcass. High numbers of first and second instars of flies were observed wandering around the body surface with C. megacephala larvae being the predominant species. Two prominent maggot masses occurred on seventh and eighth days. Bloated deterioration stage began on day eighth exposing rib bones, humerus bones and intestines. Carcass was partially sinking and the maggot masses were at the water level. On day ninth, the carcass was partially sinking and three maggot masses were observed on the exposed surface. There were very few adult flies, including a scarab beetle was sighted on the carcass at this stage. The carcass along with the maggots sunk on day tenth, leaving an oily layer on the water surface.
A Case of Zoonotic Ancylostoma ceylanicum Infection in a Suburban Area of Selangor, Malaysia

Heo CC, Rafiz AR, Ngui R

Acta Parasitol, 2021 Oct 06.
PMID: 34617193 DOI: 10.1007/s11686-021-00478-1

INTRODUCTION: The animal hookworm, Ancylostoma ceylanicum, is a dominant hookworm species of dogs and cats. However, it has increasingly been found infecting humans in Southeast Asia.
PURPOSE: We report an autochthonous case of A. ceylanicum in a suburban area of Selangor, Malaysia. A 66-year-old Indian lady who is an avid gardener presented with chronic diarrhea of 4 months' duration.
METHODS: The patient was examined clinically and colonoscopy was performed. Adult parasites obtained via colonoscopy were subjected to microscopy and molecular investigations.
RESULTS: Clinical examinations were unremarkable, and blood investigation revealed normochromic normocytic anemia. Stool occult blood was positive but negative for ova, cyst and adult parasites. Colonoscopy performed showed multiple diverticulae and worm infestation from the terminal ileum to sigmoid colon. Morphological examination on the adult worms showed the specific characteristics of Ancylostoma species. Molecular investigations further confirmed the nematode as Ancylostoma ceylanicum. She was treated with albendazole 400 mg daily for 3 days with symptomatic improvements sustained 3 months later. It is suspected that the patient had ingested or contacted soil contaminated with filariform larvae while gardening.
CONCLUSION: Information on the A. ceylanicum infection in humans, especially in urban and suburban areas, is limited, necessitating further epidemiological and clinical studies.
Fulltext Dermatitis caused by Paederus fuscipes Curtis, 1840 (Coleoptera: Staphilinidae) in student hostels in Selangor, Malaysia

Heo CC, Latif B, Hafiz WM, Zhou HZ

Southeast Asian J. Trop. Med. Public Health, 2013 Mar;44(2):197-205.
PMID: 23691629

We report a series of dermatitis cases caused by the staphilinid beetles, Paederusfuscipes Curtis, among university students staying in the residential college in Puncak Alam, Selangor, Malaysia from 1 January to 31 December 2010. A total of 360 cases (6.0%) were recorded in the Student Health Center throughout the year; the majority of patients stayed at a hostel near an oil palm plantation. Skin symptoms included erythema, edema, vesicular papules, painful blisters, burning sensation, pruritus, hyper pigmentation and peeling of skin. The commonly involved sites were the face, neck, shoulders and arms. Most students noticed the symptoms upon awakening in the morning. The patients were treated with fusidic acid cream and the symptoms resolved within 5 days. These beetles are nocturnally active and enter the room whenever a light source is available. The unintentional crushing of these beetles during sleep causes the release of its hemolymph (paederin) which is the cause of the dermatitis.
Fulltext Human pentastomiasis caused by Armillifer moniliformis in Malaysian Borneo

Latif B, Omar E, Heo CC, Othman N, Tappe D

Am J Trop Med Hyg, 2011 Nov;85(5):878-81.
PMID: 22049042 DOI: 10.4269/ajtmh.2011.11-0404

We report a case of visceral pentastomiasis caused by Armillifer moniliformis in a 70-year-old aboriginal farmer from rural Malaysian Borneo. The patient complained of upper abdominal pain, jaundice, and loss of weight. Radiological investigations and subsequent histopathological examination revealed an adenocarcinoma of the pancreas with an adjacent liver nodule containing a nymph of A. moniliformis. This report constitutes the first documented human pentastomid infection in the whole of Malaysia after nearly 40 years, and it is the third description from Malaysian Borneo. Cases of human and animal pentastomiasis in Malaysia are discussed.
Commentary on article: Aerobic microbe community and necrophagous insects associated with decomposition of pig carrion poisoned with lead by Aneyo et al. (DOI: https://doi.org/10.1016/j.legalmed.2019.101638)

Zaini NA, Ivorra T, Azmiera N, Natasha NA, Heo CC

Leg Med (Tokyo), 2021 02;48:101818.
PMID: 33271415 DOI: 10.1016/j.legalmed.2020.101818
Soil chemistry dynamics of Sus scrofa carcasses with and without delayed Diptera colonization

Heo CC, Tomberlin JK, Aitkenhead-Peterson JA

J Forensic Sci, 2021 May;66(3):947-959.
PMID: 33290606 DOI: 10.1111/1556-4029.14645

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

Heo CC, Latif B, Silahuddin SA, Nazni WA, Omar B

Trop Biomed, 2015 Mar;32(1):151-9.
PMID: 25801265

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.
Fulltext High prevalence of muscular sarcocystosis in cattle and water buffaloes from Selangor, Malaysia

Latif B, Vellayan S, Heo CC, Kannan Kutty M, Omar E, Abdullah S, et al.

Trop Biomed, 2013 Dec;30(4):699-705.
PMID: 24522140 MyJurnal

The prevalence of sarcocystosis in cattle and water buffaloes from peninsular Malaysia was investigated in abattoirs in Selangor state, February, 2011, to March, 2012. Fresh muscle samples were collected from the tongue, heart, oesophagus, diaphragm and skeletal muscles of 102 cattle and 18 water buffaloes. Each sample was initially screened by light microscopy and then fixed for further histopathological analysis. Out of 120 animals examined, 49 (40.8%) harboured the microscopic type of Sarcocystis spp. The positivity rate for cattle was 36.2% and for water buffaloes 66.7%. In cattle, the organs highly infected were the skeletal muscles and diaphragm (27% each), followed by tongue and esophagus (24.3% each), and the heart (8%). In water buffaloes, the heart was most often infected (66.7%), followed by the oesophagus (50%) and skeletal muscle (33.3%); no sarcocysts were detected in the tongue and diaphragm. The shape of the sarcocyst was fusiform to oval with a mean cyst size of 151.66 x 75.83 μm and wall thickness of 2.47 μm in cattle, and 114 x 50.81 μm cyst size and the wall thickness of 1.11 μm in water buffaloes, consistent with Sarcocystis cruzi and Sarcocystis levinei, respectively. Remaining tissue from cattle was subjected to parasite specific 18S rRNA gene PCR and Sarcocystis cruzi was confirmed, at least exemplarily. The peripheral metrocytes and the banana-shaped bradyzoites (15.23 x 2.2 μm in cattle and 11.49 x 2.45 μm in water buffalo hosts) were easily recognized. In conclusion, a high positivity rate was found in Malaysian meat-producing animals with possible implications for meat consumption and human health.
Fulltext An observation on the decomposition process of gasoline-ingested monkey carcasses in a secondary forest in Malaysia

Rumiza AR, Khairul O, Zuha RM, Heo CC

Trop Biomed, 2010 Dec;27(3):373-83.
PMID: 21399577

This study was designed to mimic homicide or suicide cases using gasoline. Six adult long-tailed macaque (Macaca fascicularis), weighing between 2.5 to 4.0 kg, were equally divided into control and test groups. The control group was sacrificed by a lethal dose of phenobarbital intracardiac while test group was force fed with two doses of gasoline LD50 (37.7 ml/kg) after sedation with phenobarbital. All carcasses were then placed in a decomposition site to observe the decomposition and invasion process of cadaveric fauna on the carcasses. A total of five decomposition stages were recognized during this study. This study was performed during July 2007. Fresh stage of control and test carcasses occurred between 0 to 15 and 0 to 39 hours of exposure, respectively. The subsequent decomposition stages also exhibited the similar pattern whereby the decomposition process of control carcasses were faster than tested one. The first larvae were found on control carcasses after 9 hours of death while the test group carcasses had only their first blowfly eggs after 15 hours of exposure. Blow flies, Achoetandrus rufifacies and Chrysomya megacephala were the most dominant invader of both carcasses throughout the decaying process. Diptera collected from control carcasses comprised of scuttle fly, Megaselia scalaris and flesh fly, sarcophagid. We concluded that the presence of gasoline and its odor on the carcass had delayed the arrival of insect to the carcasses, thereby slowing down the decomposition process in the carcass by 6 hours.
Fulltext On the occurrence of Musca domestica L oviposition activity on pig carcass in peninsular Malaysia

Heo CC, Mohamad AM, Jeffery J, Kurahashi H, Omar B

Trop Biomed, 2008 Dec;25(3):252-3.
PMID: 19287365

A forensic entomological study conducted in an oil palm plantation in Tanjung Sepat, Selangor, Malaysia on 3 August 2007 revealed that a housefly, Musca domestica Linnaeus oviposited its eggs on a freshly dead pig. This finding indicated that housefly might play an important role in forensic investigation in determining post-mortem interval (PMI), although it was not yet found in human corpses or any animal carrion. This preliminary paper presented a first record of Musca domestica eggs found on animal carcass in the country.
Juvenile hormone: Production, regulation, current application in vector control and its future applications

Nur Aliah NA, Ab-Rahim S, Moore HE, Heo CC

Trop Biomed, 2021 Sep 01;38(3):254-264.
PMID: 34362868 DOI: 10.47665/tb.38.3.066

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

Nur Aliah NA, Heo CC, Noor Shafini M, Mohd Hafizi M

Trop Biomed, 2019 Sep 01;36(3):640-653.
PMID: 33597486

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.
Ticks and tick-borne pathogens in animals and humans in the island nations of Southeast Asia: A review

Sharifah N, Heo CC, Ehlers J, Houssaini J, Tappe D

Acta Trop, 2020 Sep;209:105527.
PMID: 32447029 DOI: 10.1016/j.actatropica.2020.105527

Ticks are blood-feeding ectoparasites and major vectors of pathogens that cause infectious diseases in humans and animals worldwide including mammals, birds and reptiles. Despite the growing scientific effort in the 20th century, there is still limited information on ticks and tick-borne pathogens in Southeast Asia, especially concerning medical, veterinary, socioeconomic and agricultural aspects in the island nations. This review provides an overview of the current state of knowledge of ticks and their pathogens in the island nations of Southeast Asia and peninsular Malaysia. We aim to stimulate further research studies on ticks and tick-borne pathogens of human and veterinary importance in this geographical region.
Soft ticks (Acari: Argasidae) in the island nations of Southeast Asia: A review on their distribution, associated hosts and potential pathogens

Kazim AR, Houssaini J, Ehlers J, Tappe D, Heo CC

Acta Trop, 2021 Nov;223:106085.
PMID: 34389326 DOI: 10.1016/j.actatropica.2021.106085

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.

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