A forensic entomological study was conducted using monkey carcasses (Macaca fascicularis Raffles) that were placed in either an outdoor or indoor environment at a coastal area in Tanjung Sepat, Selangor, Malaysia during May until August 2008. We collected pupae of Chrysomya rufifacies (Marquart) from the carcasses and kept them individually. The emergence of 13 parasitic microhymenopteran, from one of the pupae occurring within a week were identified as Exoristobia philippinensis Ashmead (Hymenoptera: Encyrtidae). Another observation was made whereby a pupa of C. rufifacies was predated by a muscid larva, Ophyra spinigera (Stein). The larva squeezed into the pupa and consumed the contents. This paper report C. rufifacies as a new host record for E. philippinensis in Malaysia and highlighted the predatory behavior of O. spinigera larva in natural environment.
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.
A study on insect succession of monkey carcass in a forested area in Ulu Gombak, Selangor, Malaysia was conducted from 9 May to 18 June 2007. The third instar of the housefly, Musca domestica (Linnaeus) (Diptera: Muscidae) were only found on dry stage of a decomposed (Day-33) monkey carcass (Macaca fascicularis Raffles). This observation revealed that M. domestica maggots were found together with other muscid fly maggots, Hydrotaea (=Ophyra) spinigera (Stein) (Diptera: Muscidae) on dry stage of a carcass. However, the role of M. domestica on forensic entomological study remains unknown. This study recorded the first finding of M. domestica maggots on primate carcass in Malaysia.
Hypopygiopsis violacea, a species of fly of forensic importance, was recovered from a corpse and described for the first time. The morphological structures of the second and third instar larvae of four specimens were examined using light microscope. Observations were focused on three main morphological characters: cephalopharyngeal skeleton, anterior and posterior spiracles. Cephalopharyngeal skeleton of second instar larva is darkly pigmented and without accessory sclerite below the mouth hook. The anterior spiracles of second and third instar larvae have 8-9 papillae each, arranged in a single row. The posterior spiracle of second instar larva has two spiracular slits with no thickening of peritreme. This differentiates it from the third instar, whereby the latter has three slits for each posterior spiracle. Cephalopharyngeal skeleton of third instar larva is heavily pigmented. An accessory sclerite is found below the hook part of third instar larva but is absent in second instar. Peritreme of the posterior spiracle of third instar larva is thick almost complete encircling a button. The intersegmental spines of the cuticular surface are dome-shaped and unicuspid. Third instar larva of this species is large with size approximately 15 mm long. These findings provide important identification features of immature stages of Hy. violacea which could be useful in forensic entomology.
A human corpse at an advanced stage of decomposition was found in a house in the residential area of Bukit Mertajam, Penang, Malaysia. Entomological specimens were collected during the post-mortem and the live specimens were subsequently reared at room temperature. The time of death was estimated to have been 14 days previous to the discovery of the body based on the police investigation. Both adult and larvae of the beetle Dermestes ater (De Geer) were found to be infesting the corpse and from the stage of decomposition of the body and the estimated time of death it would appear that infestation may have begun at a relatively early stage of decomposition.
This study was carried out in an oil palm plantation in Bandar Baharu, Kedah using monkey carcasses and focuses in documenting the decomposition and dipteran colonization sequences in 50 days. This is the first study of Diptera associated with the exploitation of carcasses conducted in the north of peninsular Malaysia during the dry and wet seasons thereat. During the process of decomposition in both seasons, five phases of decay were recognized namely fresh, bloated, active decay, advance decay and dry remain. In this decomposition study, biomass loss of carcass occurred rapidly during the fresh to active decay stage due to the colonization and feeding activity of the Diptera larvae. The duration of the fresh and bloated stages of decay were the same in wet and dry seasons but later stages of decay were markedly shorter during the wet season. Twenty one species of adult Diptera were identified colonizing carcasses in the study period. Among the flies from the family Calliphoridae, Chrysomya megacephala Fabricius and Chrysomya nigripes Aubertin were recognized as the earliest arrivals on the first day of exposure. Adult Ch. nigripes was abundant for approximately two weeks after placement of the carcasses. By comparing the percentages of adults collected during the study period, the calliphorids abundance in percentages in wet season was 50.83%, but in dry season, the abundance was only about 35.2%. In contrast, the percentage of Sphaeroceridae in wet season was only 3.33%, but in the dry season, the abundance was 20.8%. Dipteran in family Phoridae, Piophilidae, Sepsidae, Drosophilidae and Dolichopodidae colonized the carcasses for a long period of time and were categorized as long term colonizers.
The mitochondiral DNA region encompassing the cytochrome oxidase subunit I (COI) and cytochrome oxidase subunit II (COII) genes of two Malaysian blow fly species, Chrysomya megacephala (Fabricius) and Chrysomya rufifacies (Macquart) were studied. This region, which spans 2303bp and includes the COI, tRNA leucine and partial COII was sequenced from adult fly and larval specimens, and compared. Intraspecific variations were observed at 0.26% for Ch. megacephala and 0.17% for Ch. rufifacies, while sequence divergence between the two species was recorded at a minimum of 141 out of 2303 sites (6.12%). Results obtained in this study are comparable to published data, and thus support the use of DNA sequence to facilitate and complement morphology-based species identification.
Myospila pudica pudica (Stein, 1915) (Diptera: Muscidae) was recorded for the first time in Malaysia during a forensic entomological study conducted at a forested area of Forensic Science Simulation Site, Faculty of Allied Health Sciences Universiti Kebangsaan Malaysia, Bangi, Selangor. This species can be differentiated from other species of its genus by having R1 setulose on dorsal surface and R4+5 more or less setulose dorsally and ventrally. The legs, including tarsi, are testaceous yellow and palpi blackish. Lateral and ventral surface of scutellum bare below the level of bristles and the third antennal segment is brownish yellow. Other features including the diverging of inner margin of lower squama from scutellum margin. This is also the first report on the occurence of M. pudica pudica (Stein, 1915) on animal carcass.
Ovitrap surveillance was initiated for eight continuous weeks to determine the distribution and abundance of Aedes sp. mosquitoes in the University of Malaya campus, Kuala Lumpur, and the impact of meteorological conditions on the Aedes populations. Two study areas within the campus were selected: Varsity Lake and Seventh Residential College. The abundance of Aedes populations in Varsity Lake was indicated by ovitrap index (OI) which ranged from 60.00%-90.00%. The mean number of larvae per ovitrap of Aedes albopictus in Varsity Lake ranged from 11.23+/-2.42-43.80+/-6.22. On the other hand, the outdoor OI for Seventh Residential College ranged from 73.33%-93.33%, respectively, while the mean number larvae per ovitrap for this area ranged from 19.33+/-4.55-35.27+/-5.46, respectively. In addition, the indoor OI of Seventh Residential College ranged from 0.00%-30.00%, while the mean number of larvae per ovitrap for Ae. albopictus ranged from 0-5.90+/-3.55. There was no significant difference (p>0.05) of Ae. albopictus population between Varsity Lake and Seventh Residential College. The studies showed a correlation between OI and mean number of larvae per ovitrap for outdoor Ae. albopictus populations in Varsity Lake and Seventh Residential College (r=0.794). There was also a correlation between the mean larvae number per ovitrap of Ae. albopictus obtained from eight weeks indoor ovitrap surveillance in Seventh Residential College with rainfall (r=0.584). However, there was no correlation between the mean larvae number per ovitrap of Ae. albopictus in both study areas with temperature and relative humidity. Aedes aegypti mosquitoes were found neither indoor nor outdoor in both study areas. This study indicated that the principal dengue vector in the university campus was most likely Ae. albopictus.
Larvae of the Synthesiomyia nudiseta (Wulp) were collected from a decomposed human corpse at the Department of Forensic Medicine, Penang Hospital. A colony of this species was established and the eggs were collected for rearing. The developmental times, rearing temperatures, and relative humidity were recorded twice daily from the time the eggs collected until adult emergence. An average of 5 larvae were randomly collected from the rearings twice daily, warm-water killed and preserved in Kahle's solution. The larval instar stages were determined by observing the number of posterior spiracular slits and the length of the preserved larvae were measured. When the larval life cycle was completed, the accumulated developmental times were calculated. A total of 8 replicates were carried out. The temperature of the rearing room was 28.5+/-1.5 degrees Celcius while the relative humidity was within 67-85%. The total developmental time for S. nudiseta was 322+/-19 hours (13.4+/-0.8 days).
The bioefficacy of nine commercial formulations of temephos against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae were evaluated in the laboratory. WHO larval bioassay with operational dosage of temephos at 1 mg/L was performed. The larval mortality was recorded every 5 minutes until complete mortality was achieved. All formulations of temephos exhibited various toxicity level against Ae. aegypti, Ae. albopictus and Cx. quinquefasciatus. Generally, larvae of Cx. quinquefasciatus was susceptible to all formulations of temephos, followed by Ae. aegypti and Ae. albopictus.
The natural and artificial mating of laboratory bred Aedes albopictus and transgenic Aedes aegypti RIDL-513A-Malaysian strain was conducted. The experiment consisted of crossmating of homologous Ae. aegypti RIDL female symbol X Ae. aegypti RIDL male symbol and reciprocal Ae. aegypti RIDL female symbol X Ae. albopictus WT male symbol. The other set comprised homologous Ae. albopictus WT female symbol X Ae. albopictus WT male symbol and reciprocal Ae. albopictus WT female symbol X Ae. aegypti RIDL male symbol. This study demonstrated that reproductive barriers exist between these two species. Cross insemination occurred between A. albopictus male and Ae. aegypti female and their reciprocals. There was 26.67% and 33.33% insemination rate in Ae. aegypti RIDL female cross-mating with A. albopictus WT male and Ae. albopictus female cross-mating with Ae. aegypti RIDL male, respectively. There was 0% hatchability in both directions of the reciprocals. There was also no embryonation of these eggs which were bleached. Although none of the female Ae. albopictus WT was inseminated in the cross-mating with Ae. albopictus WT female symbol X Ae. aegypti RIDL male symbol, a total of 573 eggs were obtained. The homologous mating was very productive resulting in both high insemination rate and hatchability rates. Generally there was a significantly higher insemination rate with artificial mating insemination of homologous than with artificial mating of reciprocal crosses. Interspecific mating between Ae. aegypti RIDL and Ae. albopictus wild type was not productive and no hybrid was obtained, indicating absence of horizontal transfer of introduced RIDL gene in Ae. aegypti to Ae. albopictus.
Larvae and adults of Culex quinquefasciatus were used for the test undertaken for malathion resistant strain (F61 - F65) and permethrin resistant strain (F54 - F58). The results showed that the LC50 for both malathion (F61 - F65) and permethrin (F54 - F58) resistant Cx. quinquefasciatus increased steadily throughout the subsequent five generations, indicating a marked development of resistance. The adult female malathion resistant strain have developed a high resistance level to malathion diagnostic dosage with a resistance ratio of 9.3 to 17.9 folds of resistance compared with the susceptible Cx. quinquefasciatus. Permethrin resistance ratio remained as 1.0 folds of resistance at every generation. It was obvious that malathion resistance developed at a higher rate in adult females compared to permethrin. Enzyme-based metabolic mechanisms of insecticide resistance were investigated based on the biochemical assay principle. From the results obtained obviously shows that there is a significant difference (p < 0.05) in esterase level in both malathion and permethrin selected strains. Female malathion selected strain has the higher level of esterase activity compared to the female permethrin selected strain at (0.8 to 1.04) alpha-Na micromol/min/mg protein versus (0.15 to 0.24) alpha-Na micromol/min/mg protein respectively. This indicated increased level of non-specific esterase is playing an important role in resistance mechanism in female malathion selected strain. Permethrin selected strain exhibited non-specific esterase activity at a very low level throughout the different life stages compared to malathion selected strain. This study suggests that life stages play a predominant role in conferring malathion and permethrin resistance in Cx. quinquefasciatus.
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.
This study was conducted to examine the effect of malathion on the development of Chrysomya megacephala. A total of 12 adult Sprague-Dawley rats was divided into 4 groups. Each animal in the 4 groups was given orally 0 (control), 10, 25 and 50ml/kg body weight of malathion, respectively. Chrysomya megacephala larvae were then allowed to grow on the liver of carcass. Larvae development was estimated by means of weight and length, time of adult emergence and survival rate. Results indicated that for the first 6 to 30 hours, larvae from control group developed more rapidly than larvae feeding on tissue containing malathion. However, the 3 doses of malathion did not exhibit significant impact on larvae length and weight. The time required for adult emergence was significantly greater for malathion-treated colony which was 10 days compared to 7 days in control colony. Control larvae of C. megacephala had higher survival rate compared to larvae exposed to the three different doses of malathion. Analysis of the tissues indicated that all rats and fly samples were positive for malathion. Malathion concentration was highest in liver. It was concluded that the presence of malathion altered the development rate of C. megacephala and thus disrupted normal postmortem interval estimation.
Matched MeSH terms: Larva/drug effects; Larva/growth & development
Piophila casei (Linnaeus) (Diptera: Piophilidae) is reported from human cadavers in two separate forensic cases for the first time in Malaysia. Both bodies were found indoors. The first case, was that of a male of unknown nationality and age and also contained maggots of the muscid Ophyra spinigera (Stein). The second case was a female Chinese whose body also contained other species of maggots but these were not identifiable.
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.
Insects found associated with corpse can be used as one of the indicators in estimating postmortem interval (PMI). The objective of this study was to compare the stages of decomposition and faunal succession between a partially burnt pig (Sus scrofa Linnaeus) and natural pig (as control). The burning simulated a real crime whereby the victim was burnt by murderer. Two young pigs weighed approximately 10 kg were used in this study. Both pigs died from pneumonia and immediately placed in an oil palm plantation near a pig farm in Tanjung Sepat, Selangor, Malaysia. One pig was partially burnt by 1-liter petrol while the other served as control. Both carcasses were visited twice per day for the first week and once thereafter. Adult flies and larvae on the carcasses were collected and later processed in a forensic entomology laboratory. Results showed that there was no significant difference between the rate of decomposition and sequence of faunal succession on both pig carcasses. Both carcasses were completely decomposed to remain stage after nine days. The species of flies visiting the pig carcasses consisted of blow flies (Chrysomya megacephala, Chrysomya rufifacies, Hemipyrellia ligurriens), flesh fly (Sarcophagidae.), muscid fly (Ophyra spinigera), soldier fly (Hermetia illucens), coffin fly (Phoridae) and scavenger fly (Sepsidae). The only difference noted was in the number of adult flies, whereby more flies were seen in the control carcass. Faunal succession on both pig carcasses was in the following sequence: Calliphoridae, Sarcophagidae, Muscidae, Phoridae and lastly Stratiomyidae. However, there was overlap in the appearance of members of these families. Blowflies continued to oviposit on both carcasses. Hence postmortem interval (PMI) can still be estimated from the partially burnt pig carcass.
The inhibitory activity of diflubenzuron, a chitin synthesis inhibitor, on the ecdysis of Aedes sp. larvae was evaluated in earthen jars and automobile tires. Two formulations of diflubenzuron were used in this study: Dimilin(R) WP (wettable powder), 25% and Dimilin GR (granular), 2%. The equivalent rate of 25 g/ha, 50 g/ha and 100 g/ha active ingredients for both WP and GR formulations were used in this study. Generally, at the higher dosage of 100 g/ha, both formulations were more effective against Aedes mosquitoes. On the whole, the WP formulation appeared to perform better than the GR formulation in terms of residual activity.
The standard laboratory strain was found to be heterozygous for susceptibility. Hence, an attempt was made to obtain a homozygous susceptible strain in Culex quinquefasciatus (Say) using single raft sib-selection method. Lab-bred females of Cx. quinquefasciatus from insectariums, Unit of Medical Entomology were used in the experiment. After blood feeding Cx. quinquefasciatus mosquitoes laid eggs in raft form, ten rafts selected randomly for the test. Each egg raft was introduced into a plastic tray from number one to number ten. Twenty-five third stage larvae from each tray were exposed to 17.5 microl from 500mg/l malathion in a paper cup label number 1 to number ten. In the bioassay, which had 100% mortality, the respective larva in that particular tray was bred to adult stage for the following generation. Less than 7days old female mosquitoes that emerged from F(0) were used in the test. The F(0) and the subsequent adult and larval stage generations were subjected to adult and larval bioassay. After selection for about 10 generations, a homozygous susceptible strain in Cx. quinquefasciatus was obtained.