A number of methods have been used for the detection of the presence of microsarcocysts in animals, but little information exists on the value between the various methods. This study therefore examined for Sarcocystis spp. using three different methods in 105 samples of skeletal muscle collected from goats slaughtered in an abattoir in Selangor, Malaysia from January to February 2014. Three methods were used, direct light microscopy of squashed fresh muscle tissues; histological examination of fixed, sectioned, and hematoxylin and eosin (H&E)-stained samples of muscle; and molecular identification by polymerase chain reaction (PCR). Of the 105 tissue samples, 55 (52.38 %) were positive by light microscopy (LM), 46 (43.8 %) by histology, and 95 (90.48 %) by PCR. Only 29 (27.6 %) and 5 (4.76 %) samples were positive and negative, respectively, by all three methods. The cysts were elongated to a spindle shape with a mean size of 393.30 × 81.6 μm and containing banana-shaped bradyzoites of size 12.32 × 2.08 μm. The wall of the cyst was radially striated with a thickness of 2.83 μm. Samples were tested for the presence of Sarcocystis-specific 18S rRNA and were identified as Sarcocystis capracanis. Of the three methods used, the PCR test appears to be the most useful method for the diagnosis of sarcocystosis especially for species identification.
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.
A Trypanosoma screening was conducted on 130 pools comprising 1,241 ticks, collected from 674 selected farm ruminants in Peninsular Malaysia. Of these, nine pools were tested positive for Trypanosoma. Subsequent BLAST searches revealed that the 18S rRNA gene sequences were closely related to Trypanosoma rhipicephalis isolate Chaco CB, with percentage similarities ranging from 95.56% to 99.84%. Phylogenetic analysis showed that three of the nine sequences formed a clade with Trypanosoma rhipicephalis. The remaining six Trypanosoma sequences formed a distinct clade, separate from T. rhipicephalis and other Trypanosoma species, with genetic distances of 4.34% and 4.33-4.58%, respectively. This study marks the first report of tick-associated Trypanosoma in Malaysia and underscores significant research gaps regarding trypanosome interactions with tick hosts in the region.
To shed light on the importance of tick-borne diseases, especially in farm animals that often contact with farm workers, this study aimed to identify ticks and tick-borne pathogens in ruminants in Malaysia. Accordingly, specimen collection was conducted across Peninsular Malaysia yielded a total of 1241 ticks collected from 674 farm ruminants. Among these, four tick species were identified, with Rhipicephalus microplus being the most prevalent, constituting 99.03 % of the total tick population. Analysis of 130 tick pools revealed three positives for Borrelia. BLAST analyses of the flaB and 16S rRNA genes revealed high similarities to Borrelia theileri, ranging from 98.78 to 100 % for flaB and 99.23-99.45 % for 16S rRNA. These results align with the phylogenetic trees, where sequences from both genes clustered together with B. theileri, further supporting this identification. No Rickettsia and Bartonella bacteria were detected. This study represents the first occurrence of B. theileri in R. microplus in Malaysia.
Rhipicephalus haemaphysaloides is endemic in South and Southeast Asia, and it is known to parasitise domestic animals such as cattle, horses, sheep, and dogs. Despite infestation on multiple hosts, little attention has been paid to its morphological and molecular variation. The present study describes local morphological abnormalities for the first time in R. haemaphysaloides from Malaysia, and highlights the presence of three cytochrome c oxidase subunit I (COI) operational taxonomic units with 0.46-9.81% distances within the so-called R. haemaphysaloides in Malaysia, Thailand, and Pakistan.
A total of 68 adult hard ticks were recovered from 5 cases of infestation involving pet rabbits (Oryctolagus cuniculus domesticus) in Malaysia. The ticks were later identified as the tropical brown dog tick, Rhipicephalus linnaei (Audouin, 1826). Pathogen screening showed 2 out of the 68 Rh. linnaei ticks (2.94% infection rate) were positive for Rickettsia. BLAST analyses revealed that both samples were identical to Rickettsia conorii subsp. raoultii. This paper reports the first detection of R. conorii subsp. raoultii in Rh. linnaei, also highlighting the potential zoonotic threat posed by ticks infesting exotic pets such as rabbits.
A partially skeletonized human corpse was found in bushes in Selangor, Malaysia in June 2020. Entomological evidence was collected during the autopsy and sent to the Department of Medical Microbiology and Parasitology, Faculty of Medicine, Universiti Teknologi MARA (UiTM) for minimum postmortem interval (PMImin) analysis. Standard protocols were applied when processing preserved and live insect specimens of both larval and pupal stages. Entomological evidence revealed that the corpse was colonized by Chrysomya nigripes Aubertin, 1932 (Diptera: Calliphoridae) and Diamesus osculans (Vigors, 1825) (Coleoptera: Silphidae). Chrysomya nigripes was chosen as the PMImin indicator as this fly species is an earlier colonizer compared to D. osculans beetle larvae which their presence is the indicative of late stage of decomposition. For the present case, the pupae of C. nigripes were the oldest insect evidence collected and based on the available developmental data, the estimated minimum PMI was established between 9 and 12 days. It is noteworthy to highlight that this is the first record of D. osculans colonization on a human corpse.
A basic tenet of forensic entomology is development data of an insect can be used to predict the time of colonization (TOC) by insect specimens collected from remains, and this prediction is related to the time of death and/or time of placement (TOP). However, few datasets have been evaluated to determine their accuracy or precision. The black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) is recognized as an insect of forensic importance. This study examined the accuracy and precision of several development datasets for the black soldier fly by estimating the TOP of five sets of human and three sets of swine remains in San Marcos and College Station, TX, respectively. Data generated from this study indicate only one of these datasets consistently (time-to-prepupae 52%; time-to-eclosion 75%) produced TOP estimations that occurred within a day of the actual TOP of the remains. It is unknown if the precolonization interval (PreCI) of this species is long, but it has been observed that the species can colonize within 6 d after death. This assumption remains untested by validation studies. Accounting for this PreCI improved accuracy for the time-to-prepupae group, but reduced accuracy in the time-to-eclosion group. The findings presented here highlight a need for detailed, forensic-based development data for the black soldier fly that can reliably and accurately be used in casework. Finally, this study outlines the need for a basic understanding of the timing of resource utilization (i.e., duration of the PreCI) for forensically relevant taxa so that reasonable corrections may be made to TOC as related to minimum postmortem interval (mPMI) estimates.
The use of drones has rapidly evolved over the past decade involving a variety of fields ranging from agriculture, commercial and becoming increasingly used in disaster management or humanitarian aid. Unfortunately, the evidence of its use in mass disasters is still unclear and scarce. This article aims to evaluate the current drone feasibility projects and to discuss a number of challenges related to the deployment of drones in mass disasters in the hopes of empowering and inspiring possible future work. This research follows Arksey and O'Malley framework and updated by Joanna Briggs Institute Framework for Scoping Reviews methodology to summarise the results of 52 research papers over the past ten years, from 2009 to 2020, outlining the research trend of drone application in disaster. A literature search was performed in Medline, CINAHL, Scopus, individual journals, grey literature and google search with assessment based on their content and significance. Potential application of drones in disaster are broad. Based on articles identified, drone application in disasters are classified into four categories; (1) mapping or disaster management which has shown the highest contribution, (2) search and rescue, (3) transportation and (4) training. Although there is a significant increase in the number of publications on use of drone in disaster within the last five years, there is however limited discussion to address post-disaster healthcare situation especially with regards to disaster victim identification. It is evident that drone applications need to be further explored; to focus more on drone assistance to humans especially in victim identification. It is envisaged that with sufficient development, the application of drones appears to be promising and will improve their effectiveness especially in disaster management.
Signal fly, Scholastes sp. (Diptera: Platystomatidae) was observed associated with animal carcasses in Malaysia. The first observation was on a monkey carcass, which was killed by using a handgun and immediately placed in a forested area in Gombak, Selangor while the second observation was on a pig that died of natural causes and whose carcass was placed in an oil palm plantation in Tanjung Sepat, Selangor. Both animal carcasses were visited by Scholastes sp. flies during the fresh decomposition period. However, the role Scholastes flies in the decomposition process remains unknown. In this paper, we report the occurrence of Scholastes sp. on animal carcasses in Malaysia for the first time.
In medical, veterinary and forensic entomology, the ease and affordability of image data acquisition have resulted in whole-image analysis becoming an invaluable approach for species identification. Krawtchouk moment invariants are a classical mathematical transformation that can extract local features from an image, thus allowing subtle species-specific biological variations to be accentuated for subsequent analyses. We extracted Krawtchouk moment invariant features from binarised wing images of 759 male fly specimens from the Calliphoridae, Sarcophagidae and Muscidae families (13 species and a species variant). Subsequently, we trained the Generalized, Unbiased, Interaction Detection and Estimation random forests classifier using linear discriminants derived from these features and inferred the species identity of specimens from the test samples. Fivefold cross-validation results show a 98.56 ± 0.38% (standard error) mean identification accuracy at the family level and a 91.04 ± 1.33% mean identification accuracy at the species level. The mean F1-score of 0.89 ± 0.02 reflects good balance of precision and recall properties of the model. The present study consolidates findings from previous small pilot studies of the usefulness of wing venation patterns for inferring species identities. Thus, the stage is set for the development of a mature data analytic ecosystem for routine computer image-based identification of fly species that are of medical, veterinary and forensic importance.
The present study aims to investigate the susceptibility status of Aedes albopictus, Culex quinquefasciatus, and Cx. vishnui collected from a pig farm in Tanjung Sepat, Selangor, toward 11 insecticides representing the classes of organochlorines, carbamates, organophosphates, and pyrethroids. The results of a World Health Organization adult mosquito bioassay revealed that Ae. albopictus, Cx. quinquefasciatus, and Cx. vishnui exhibited different susceptibility toward various insecticides. Overall, pyrethroids were able to induce rapid knockdown for all test mosquito species. The pyrethroids lambdacyhalothrin and etofenprox were able to cause high mortality (> 80%) of all 3 species. The findings of the present study will benefit local authorities in selecting appropriate dosage of insecticides to be used in mosquito control in this area.
Nymphal and larval Amblyomma cordiferum ticks, a relatively rare species, were collected from the Malaysian house rat (Rattus rattus diardii) in Peninsular Malaysia. Redescription and molecular analysis of nymphs and larvae, based on the 12S rRNA, 16S rRNA, and COI genes, revealed divergence from A. cordiferum in Taiwan, suggesting that the Taiwanese tick specimens may represent a different taxon. Molecular analysis of the pathogens in these specimens revealed three sequences of Rickettsia closely related or identical to Rickettsia raoultii (99.71-100%), two sequences of relapsing fever Borrelia identical to Borrelia theileri, and four sequences of Bartonella identical to Bartonella phoceensis. This study also identifies a new host record for A. cordiferum in R. r. diardii and reports the first detection of Rickettsia, Borrelia, and Bartonella in this tick species.
An undescribed relapsing fever group Borrelia species was detected in a male Haemaphysalis semermis tick infesting a rural cat in an indigenous population in Pahang National Park, Peninsular Malaysia. The 16 S rRNA gene sequence revealed close similarity of this variant to several undescribed Borrelia species and Borrelia theileri, with genetic distances ranging from 0.58 to 0.72%. Furthermore, the flaB, gyrB, and the concatenated 16 S rRNA + flaB + gyrB sequence analyses demonstrated that this variant is distinctly separated from multiple undescribed Borrelia species, Borrelia miyamotoi, and B. theileri, with genetic distances ranging from 3.41 to 7.00%. This study not only reports the first Borrelia found in H. semermis but also suggests that it forms a distinct clade within the relapsing fever group in Peninsular Malaysia.
Manual ground searches and cadaver dogs are traditional methods for locating remains, but they can be time- and resource-intensive, resulting in the decomposition of bodies and delay in victim identification. Therefore, thermal imaging has been proposed as a potentially useful tool for detecting remains based on their temperature. This study investigated the potential of a novel search technique of thermal drones to detect surface remains through the detection of maggot mass temperatures. Two trials were carried out at Selangor, Malaysia, each utilizing 12 healthy male Oryctolagus cuniculus European white rabbits and DJI Matrice 300 RTK drone China, equipped with a thermal camera; Zenmuse H20T to record the thermal imaging footage of the carcasses at various heights (15, 30, 60-100 m) for 14 days for each trial. Our results demonstrated that the larval masses and corresponding heat emissions were at their largest during the active decay stage; therefore, all the carcasses were observable in thermal images on day 5 and remained until day 7. Statistical analyses showed that (1) no statistically significant differences in thermal images between clothed and unclothed subjects (p > 0.05); (2) 15 m above ground level was proven to be the optimal height, as it showed the greatest contrast between the carcass heat signature and the background (p
Disaster Victim Identification (DVI) following mass fatality events is critical in bringing closure to the victims' family members and their loved ones. However, post-disaster environments are typically unfavourable and pose difficulties for the execution of DVI procedures. Delays in collecting, transporting, and processing the samples may cause DNA to degrade, adversely impacting the identification process. Therefore, this study aimed to demonstrate the co-utilisation of RapidHIT ID (RHID) and DJI Matrice 300 (M300) for onsite DNA analysis using buccal swab samples. 40 samples (two replicates) were collected from 20 "victims" and another 40 (two replicates) from 20 corresponding "relatives". The first replicates were processed using RHID (n = 40) and the second replicates were analysed via conventional technologies (n = 40). This paper observed the genotyping success rate, kinship matching, concordance, comparing different sample collectors, storage interval, and time taken for both procedures. Results of this study showed that RHID could generate DNA profiles for all the samples (n = 40) with 90% of them showing full profiles, and managed to process samples that have been stored for up to six months. The drone-assisted procedure exhibits less time to obtain and analyse the samples but can still produce DNA profiles concordant with the conventional method (p > 0.05). In conclusion, RHID is sufficient to generate interpretable DNA profiles in harsh environments, and transporting samples by M300 drone can reduce the exposure time to process more quality DNA for DVI.