Mammal diversity assessments based on DNA derived from invertebrates have been suggested as alternatives to assessments based on traditional methods; however, no study has field-tested both approaches simultaneously. In Peninsular Malaysia, we calibrated the performance of mammal DNA derived from blowflies (Diptera: Calliphoridae) against traditional methods used to detect species. We first compared five methods (cage trapping, mist netting, hair trapping, scat collection, and blowfly-derived DNA) in a forest reserve with no recent reports of megafauna. Blowfly-derived DNA and mist netting detected the joint highest number of species (n = 6). Only one species was detected by multiple methods. Compared to the other methods, blowfly-derived DNA detected both volant and non-volant species. In another forest reserve, rich in megafauna, we calibrated blowfly-derived DNA against camera traps. Blowfly-derived DNA detected more species (n = 11) than camera traps (n = 9), with only one species detected by both methods. The rarefaction curve indicated that blowfly-derived DNA would continue to detect more species with greater sampling effort. With further calibration, blowfly-derived DNA may join the list of traditional field methods. Areas for further investigation include blowfly feeding and dispersal biology, primer biases, and the assembly of a comprehensive and taxonomically-consistent DNA barcode reference library.
Considering that crimes against animals such as illegal killing and cruelty have been alarmingly increasing and since burning is one of the common ways for disposing cadavers, ability to estimate minimum postmortem interval (PMI) using entomological data merits consideration. Chrysomya megacephala and Chrysomya rufifacies are common necrophagous species recovered from cadavers in many countries including Malaysia. Specific studies focusing on the oviposition and developmental patterns of both species on cadavers manifesting different levels of burn as described by the Crow-Glassman Scale (CGS) remain scarce. In four replicates, rabbit carcasses were burned to CGS levels #1, #2 and #3 by varying the amount of petrol used and duration of burning. Oviposition by C. megacephala and C. rufifacies was delayed by one day in the case of carcasses burned to the CGS level #3 (p<0.05) when compared with that of controls. Such delay in oviposition was not observed in the CGS level #1 and #2 carcasses. No significant differences (p>0.05) in the duration of development were found between control and burned carcasses. These findings deserve consideration while estimating minimum PMI since burning as a mean for disposing animal and human cadavers is gaining popularity.
Hermetia illucens larvae by nature are a decomposer which fed on organic wastes. This study explores the potential of producing biodiesel using lipids from H. illucens larvae. Three types of organic wastes (sewage sludge, fruit waste and palm decanter cake from oil palm mill) were selected based on considerable generation and disposal concern in the area of study as well as lack of investigations as feed for Hermetia illucens larvae in current literatures. Growth rate of the larvae was determined with studying the changes in the biomass per day. H. illucens larvae fed with fruit waste and palm decanter cake have shown growth rates of 0.52±0.02 and 0.23±0.09 g d(-1), respectively. No positive sign of growth were observed in the larvae fed with treated sewage sludge (-0.04±0.01 g d(-1)). Biodiesel as fatty acid methyl ester (FAME) was synthesized by transesterification of the larvae lipid using sulphuric acid as catalyst in methanol. FAME produced was ascertained using ATR-FTIR spectroscopy and GC-MS. The main compositions of fatty acid were found to be C12:0, C16:0 and C18:1n9c. Fatty acid composition of C12:0 fed with fruit waste, sewage sludge and palm decanter was found to be most abundant in the larvae lipid. The amount of C12:0 obtained was 76.13%, 58.31% and 48.06%, respectively. In addition, fatty acid of C16:0 was attained at 16.48% and 25.48% fed with sewage sludge and palm decanter, respectively. Based on the findings, FAME derived from larvae lipids is feasible to be used for biodiesel production.
Megaselia scalaris (Loew) is a cosmopolitan polyphagous small fly with the ability of exploiting variety of ecological niches. Different life history stages act as detritivore, parasite, and parasitoid of wider spectrum of plant and animal matter under natural and laboratory conditions. Here, for the first time we present the opportunistic parasitism of M. scalaris on Otobius megnini, which act as a vector of Q fever and is capable of causing paralysis, toxic conditions, otoacariasis and otitis in humans and other animals. Tick samples from the ear canals of 14 thoroughbred horses were brought to the laboratory and several days later, larvae of M. scalaris were found feeding on immature stages of O. megnini. When the development was completed pupae were found attached to adult ticks and all nymphs were found dead. This context reveals the capability of M. scalaris surviving on O. megnini and the risk of their invading ear canals of horses.
Paraquat poisoning is commonly associated with suicide or homicide in Malaysia. In a case involving advanced body decomposition, pathological analysis regarding the cause of death may become difficult or almost impossible. Insects serve as common alternative matrix for poison detection in forensic analysis. Paraquat detection via secondary bioaccumulation in fly larvae tissue has never been reported. In this study, tissues from blowfly larvae collected from a rabbit carcass with paraquat poisoning were analyzed for secondary bioaccumulation. Larvae samples were collected and analyzed using liquid-liquid extraction. The detection was performed via reduction of quaternary ammonium presence in paraquat and analyzed using gas chromatography-mass spectrometry (GC-MS) with selected ion monitoring mode (SIM mode). GC-MS showed the elution of reduced paraquat was at retention time 12.8 min. Blowfly larvae tissue has proven useful as a secondary detector in paraquat-related deaths.
In forensic entomology, larval rearing usually includes the presence of biological contaminants including scuttle flies (Diptera: Phoridae). Scuttle flies are recognized as forensically important insects and have been reported causing nuisance and contamination in laboratory environments. This paper reports for the first time the finding of multiple scuttle fly species affecting colonies of third instar larvae of the Oriental latrine blowfly, Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae), reared indoors at the Forensic Science Simulation Site, Universiti Kebangsaan Malaysia. Adult scuttle flies were discovered inside a rearing container after the emergence of adult C. megacephala., The scuttle fly species are Megaselia scalaris (Loew), M. spiracularis Schmitz and Puliciphora borinquenensis (Wheeler). Notes on the life history and biology of these species are discussed herein.
The distributions of flies are not only confined to ground level but can also be at higher altitudes. Here, we report three forensic cases involving dipterans in high-rise buildings in Kuala Lumpur, Malaysia. Case 1 involved a corpse of adult female found at the top floor of a fifteen-story apartment. Case 2 dealt with a body of a 75-year-old female discovered in a bedroom on the eleventh floor of an eighteen-story building, while Case 3 was a 52-year-old male found in his fifth floor shop house. Interestingly, entomological analysis revealed that all corpses were infested with similar Dipterans: Megaselia scalaris (Loew) (Diptera: Phoridae), Synthesiomyia nudiseta (Wulp) (Diptera: Muscidae) and sarcophagid (Diptera: Sarcophagidae). The first two species were commonly associated with corpses found indoors at ground level. We noted the additional occurrence of blowflies Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) and Chrysomya rufifacies Macquart (Diptera: Calliphoridae) larvae in Case 2 and Case 3, respectively. Findings from this study are significant as they demonstrate that certain groups of fly can locate dead bodies even in high-rise buildings. Forensic entomofauna research on corpses found at high elevation is scarce and our study has highlighted the peculiarity of the fly species involved in Malaysia.
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.
The stages of decomposition and the faunal succession on rabbit carcasses in three different habitats, namely jungle, rural, and highland areas, were studied. Three New Zealand White rabbit (Oryctolagus cuniculus) carcasses weighing ∼2 kg were sampled daily until the decomposition process was completed. Representative specimens of adult flies, larvae, pupa, and mites were collected from the carcasses and processed in the laboratory. There were differences in decomposition rate and faunal succession between the carcasses. The fastest rate of decomposition was recorded in rural area, and the slowest rate of decomposition was recorded in highland area. The carcasses exhibited the same pattern of colonization by adult flies, but the dominant species of larvae and adult flies on each carcass in specific habitats were different. The primary species of flies recorded in jungle were Chrysomya megacephala F., Chrysomya rufifacies (Macquart), Chrysomya chani Kurahashi, Chrysomya villenuevi Patton, Chrysomya nigripes Aubertin, Chrysomya pinguis (Walker), Hemipyrellia ligurriens (Wiedemann), Hemipyrellia tagaliana (Bigot), Hypopyiopsis fumipennis (Walker), Hypopygiopsis violacea (Macquart), and Hydrotaea spinigera Stein represented by both adults and larvae. Musca domestica L., Atherigona sp., Lioproctia pattoni (Senior-White), Lioproctia saprianovae Pape & Bänziger, and Seniorwhitea princeps (Wiedemann) were represented by adults only. The biodiversity of flies in the rural area were C. megacephala, C. rufifacies, H. ligurriens, Fannia canicularis L., Hydrotaea chalcogaster (Wiedemann), and Hyd. spinigera represented by both adults and larvae, meanwhile M. domestica, Atherigona sp., Boettcherisca peregrina (Robineau-Desvoidy), Parasarcophaga taenionota Wiedemann, Parasarcophaga scopariiformis Senior-White, and S. princeps were represented by adults only. The species of flies collected in the highland area were Lucilia porphyrina (Walker), C. megacephala, C. rufifacies, C. villenuevi, C. pinguis, H. ligurriens, Hyd. spinigera, Hyd. chalcogaster, F. canicularis, and Boettcherisca highlandica Kurahashi & Tan represented by both adults and larvae, whereas C. nigripes, Chrysomya thanomthini Kurahashi & Tumrasvin, M. domestica, Atherigona sp., Parasarcophaga albiceps Meigen, P. taenionota, Sepsidae, Phoridae, and Millichidae were represented by adults only. Faunal succession followed the sequence of dominant flies, i.e., Calliphoridae, Sarcophagidae, Muscidae, Sepsidae, and lastly Stratiomyidae for jungle, or Sepsidae for rural and highland studies. Mites, from suborders Mesostigmata, Prostigmata, Astigmatina, and Oribatida, were also recovered throughout decomposition, which could be used for future implementation in forensic investigations. The data obtained from this study could provide more accurate indicators for local forensic scientists in solving criminal cases especially on the determination of time and primary location of death.
Most tropical mammal species are threatened or data-deficient. Data collection is impeded by the traditional monitoring approaches which can be laborious, expensive and struggle to detect cryptic diversity. Monitoring approaches using mammal DNA derived from invertebrates are emerging as cost- and time-effective alternatives. As a step towards development of blowfly-derived DNA as an effective method for mammal monitoring in the biodiversity hotspot of Peninsular Malaysia, our objectives were (i) to determine the persistence period of amplifiable mammal mtDNA in blowfly guts through a laboratory feeding experiment (ii) to design and test primers that can selectively amplify mammal COI DNA mini-barcodes in the presence of high concentrations of blowfly DNA. The persistence period of amplifiable mammal mtDNA in blowfly guts was 24 h to 96 h post-feeding indicating the need for collecting flies within 24 h of capture to detect mammal mtDNA of sufficient quantity and quality. We designed a new primer combination for a COI DNA mini-barcode that did not amplify blowfly DNA and showed 89% amplification success for a dataset of mammals from Peninsular Malaysia. The short (205 bp) DNA mini-barcode could distinguish most mammal species (including separating dark taxa) and is of suitable length for high-throughput sequencing. Our new DNA mini-barcode target and a standardized trapping protocol with retrieval of blowflies every 24 h could point the way forward in the development of blowfly-derived DNA as an effective method for mammal monitoring.
The species of Billaea Robineau-Desvoidy (Diptera, Tachinidae) from the eastern Palearctic and Oriental regions are revised. Ten described species are recognized, viz. B. atkinsoni (Baranov) (new records for Japan, Laos, Malaysia, Pakistan and Thailand), B. ficorum (Townsend), B. fortis (Rondani), B. impigra Kolomiets (new record for China), B. kolomyetzi Mesnil, B. malayana Malloch, B. morosa Mesnil, B. robusta Malloch, B. steini (Brauer et Bergenstamm) and B. triangulifera (Zetterstedt) and nine species are described as new to science, B. brevicauda Zhang et Shima sp. nov. (China), B. carinata Zhang et Shima sp. nov. (China), B. chinensis Zhang et Shima sp. nov. (China and Vietnam), B. flava Zhang et Wang sp. nov. (China), B. kurahashii Zhang et Shima sp. nov. (Laos and Thailand), B. micronychia Zhang et Shima sp. nov. (China and Japan; previously misidentified from China as B. irrorata (Meigen)), B. papei Zhang et Shima sp. nov. (Malaysia), B. setigera Zhang et Shima sp. nov. (China) and B. verticalis Shima et Zhang sp. nov. (China). Billaea fasciata (Townsend, 1928) is treated as a junior synonym of B. ficorum (Townsend, 1916), syn. nov. Billaea irrorata is no longer recorded from the eastern Palearctic. A key to 19 species of Billaea from the eastern Palearctic and Oriental regions and 101 figures of male terminalia, bodies, heads and abdomens are given.
The Oriental, Australasian and Oceanian genus Caiusa Surcouf, 1920 is revised, species concepts being based on male and female genitalia. A key to males for all known species, and a key to females for all except one are given. All relevant types still in existence have been studied, complete synonymies given and the geographical distribution reconsidered. The eight species included in the genus are: Caiusa borneoensis sp. nov. (Malaysia, Thailand, Vietnam); Caiusa coomani Séguy, 1948 (China, Malaysia, Singapore, Thailand, Vietnam); Caiusa indica Surcouf, 1920 (Australia, Cambodia, India, Indonesia, Malaysia, Papua New Guinea, Philippines, Singapore, Solomon Islands, Sri Lanka, Thailand, Vietnam); Caiusa karrakerae sp. nov. (Malaysia, Thailand); Caiusa kurahashii sp. nov. (Indonesia, Japan, Philippines); Caiusa pooae sp. nov. (Thailand); Caiusa testacea Senior-White, 1923 (India, Nepal, Sri Lanka) and Caiusa violacea Séguy, 1925, stat. rev. (Cambodia, China, Laos, Malaysia, Taiwan, Thailand, Vietnam). A lectotype is designated for Caiusa indica to fix the interpretation of the name. Caiusa nigronitens Senior-White, 1923, syn. nov. and Caiusa surcoufi Bezzi, 1927, syn. nov. are established as junior synonyms of Caiusa indica. Caiusa violacea is correctly diagnosed and errors in the original description of the female holotype are pointed out. Caiusa dubiosa Villeneuve, 1927 is established as a junior synonym of C. violacea, syn. nov. Seven Caiusa species have been reared from the egg mass of various species of frogs. The reproductive mode of the eighth species, i.e., C. indica, is unknown. Five species, i.e., C. borneoensis, C. coomani, C. karrakerae, C. kurahashii and C. violacea have been reared from one or more of the foam nesting frog species Chiromantis nongkhorensis (Cochran, 1927), Polypedates leucomystax (Gravenhorst, 1927), Polypedates megacephalus Hallowell, 1861, Rhacophorus annamensis Smith, 1924, Rhacophorus dulitensis Boulenger, 1892, Rhacophorus kio Ohler & Delorme, 2005 and Rhacophorus owstoni (Stejneger, 1907) all belonging in the family Rhacophoridae in Anura. These five Caiusa species all have a specialised ovipositor tip, with small spine-like setae on the ST8 and the hypoproct, probably enabling the flies to oviposit on a foam nest with a hardened outer surface. They form a monophyletic group on account of these features of the ovipositor, unique in the Oestroidea. The sixth species, C. testacea, has been reared from a frog egg mass, the frog species being unknown. Its ovipositor structure is also unknown. The seventh species, C. pooae, has been reared once from the jelly-like egg mass of Feihyla hansenae (Cochran, 1927), also in Rhacophoridae. Caiusa pooae females do not have spine-like setae on the ovipositor, a fact correlated with the soft outer surface of the jelly-like egg mass on which a C. pooae female had oviposited. The extreme rarity of C. pooae oviposition on Feihyla hansenae egg masses may indicate that this fly perhaps has another, unknown, regular oviposition substrate. Caiusa pooae and C. indica make up a second monophyletic group within Caiusa. Caiusa indica, the most common and most widespread species of the genus, has an ovipositor structure similar to C. pooae. Its breeding substrate is unknown and it occurs both within and outside the distributional area of Rhacophoridae. Possibly both C. indica and C. pooae share a regular oviposition substrate that has still to be discovered. The holotype female of Plinthomyia emimelania Rondani, 1875 from Sarawak is established as a member of the genus Bengalia Robineau-Desvoidy, 1830, thus Plinthomyia Rondani, 1875 becomes a junior synonym of Bengalia Robineau-Desvoidy, 1830, syn. nov. It is removed from the synonymy of Phumosia Robineau-Desvoidy, 1830.
Trischidocera Villeneuve, 1915 includes two species, T. sauteri Villeneuve, 1915 (Taiwan and Malaysia) and T. yunnanensis Chao & Zhou, 1987 (China). The systematic placement of Trischidocera has been controversial. It was originally placed within the "Thryptoceratidae" (= "Actiidae"), then moved to Germariini, then considered an unplaced Tachinidae, and more recently placed in Ormiini. Here, the genus is revised, the type-species is redescribed and illustrated, and its systematic placement is discussed. The genus is removed from Ormiini and considered incertae sedis.
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 5-year-old boy presented with right eye pain associated with tearing and photophobia of 1-day duration. He gave a history of playing with a river crab when suddenly the crab clamped his fingers. He attempted to fling the crab off, but the crab flew and hit his right eye. Ocular examination revealed a right eye corneal ulcer with clumps of fibrin located beneath the corneal ulcer and 1.6 mm level of hypopyon. At presentation, the Seidel test was negative, with a deep anterior chamber. Culture from the corneal scrapping specimen grew Citrobacter diversus and Proteus vulgaris, and the boy was treated with topical gentamicin and ceftazidime eyedrops. Fibrin clumps beneath the corneal ulcer subsequently dislodged, and revealed a full-thickness corneal laceration wound with a positive Seidel test and shallow anterior chamber. The patient underwent emergency corneal toileting and suturing. Postoperatively, he was treated with oral ciprofloxacin 250 mg 12-hourly for 1 week, topical gentamicin, ceftazidime, and dexamethasone eyedrops for 4 weeks. Right eye vision improved to 6/9 and 6/6 with pinhole at the 2-week follow-up following corneal suture removal.
In a forensic acarology study, conducted in a secondary forest of Forensic Science Simulation Site, Universiti Kebangsaan
Malaysia (UKM) in Bangi, Selangor, six carcasses consisted of three monkeys and three rabbits were placed individually
in steel enclosures and observed for 35 days. The carcasses, the soil beneath them and pitfall traps were checked daily
for insects and mites. A species of mesostigmatid mite, Macrocheles scutatiformis was discovered and this is a first time
reported in Malaysia. There are about 94.0% of Macrocheles present inside the cages of the monkey and rabbit carcasses.
Most mites in this study were collected from soil beneath the carcasses, on a dung beetle and a Chrysomya rufifacies
larva. Our findings showed that M. scutatiformis have a potential role in forensic investigation involving dead animals
perhaps corpses especially during the dry stage of decomposition.
Scuttle flies of genus Puliciphora Dahl (Diptera: Phoridae) are recorded for the first time in Malaysia from rabbit carcasses placed in concealed environments. They consist of Puliciphora borinquenensis Wheeler ♂♀, Puliciphora obtecta Meijere ♀ and Puliciphora beckeri Meijere ♀. All species were obtained from rabbit carcasses in used luggage and garbage bin placed at Forensic Science Simulation Site, Universiti Kebangsaan Malaysia, Bangi, Selangor. The specimens were collected from the carcasses using a modified Malaise trap, an entomological aspirator and preserved in 70% ethanol. This report expands the geographical distributions of these species and their microhabitat, suggests its possible important role in forensic entomology.
Preservation of larvae retrieved from cadavers is important in ensuring the quality and integrity of entomological specimens used for the estimation of post-mortem interval (PMI). The process of killing and preserving larvae could distort the larvae leading to inaccurate estimation of PMI. In this study, the effects of killing Chrysomya megacephala larvae with hot water at different temperatures and subsequent maintenance in various preservatives were determined. Larvae not killed by hot water but preserved directly were used as control. The types of preservative used were 10% formalin, 70% ethanol and Kahle's solution. The morphological features examined were length, turgidity, curvature and coloration of larvae. Larvae killed in 80ºC hot water have shorter mean length (12.47 ± 2.86 mm) compared to those in 60ºC hot water (12.95 ± 2.69 mm). Increasing the duration of preservation in all types of preservative caused elongations of larvae treated or untreated with hot water. There were no significant changes in larval turgidity preserved in Kahle's solution compared to other two preservatives and were unaffected by the duration of storage. Larvae preserved in Kahle's solution experienced the least changes in coloration and shape compared to other preserved larvae in 70% ethanol or 10% formalin. Larvae directly immersed alive in 70% ethanol experienced the most changes in curvature, coloration and turgidity. This study suggested that killing larvae with hot water at 80ºC and preservation in Kahle's solution is the optimum method resulting in least changes in morphological features of Ch. megacephala larvae.
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.
This study investigated the influence of paraquat, a prevalent poison used by suicides, on initial oviposition and development of Chrysomya megacephala (Fabricius) using minced-beef substrates. Paraquat in lethal dose for human (40 mg/kg), two times the lethal dose (80 mg/kg) and five times the lethal dose (200 mg/kg) were mixed thoroughly with respective minced-beef substrates (1 kg each) that were decomposed in a shaded habitat fully protected from rain. Results of four replications of the above experiment revealed that the presence of paraquat neither delayed initial oviposition nor prolonged the developmental stages of C. megacephala. Therefore, estimation of postmortem interval (PMI) based on empirical baseline data obtained using animal models devoid of any poisons would still be appropriate for estimating PMI in paraquat-related deaths.