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Machine learning analysis of wing venation patterns accurately identifies Sarcophagidae, Calliphoridae and Muscidae fly species

Ling MH, Ivorra T, Heo CC, Wardhana AH, Hall MJR, Tan SH, et al.

Med Vet Entomol, 2023 Dec;37(4):767-781.
PMID: 37477152 DOI: 10.1111/mve.12682

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.

Matched MeSH terms: Sarcophagidae*
Identification of medically and forensically relevant flies using a decision treelearning method

Tanajitaree C, Sanit S, Sukontason KL, Sukontason K, Somboon P, Anakkamatee W, et al.

Trop Biomed, 2023 Mar 01;40(1):80-87.
PMID: 37356007 DOI: 10.47665/tb.40.1.019

Blow flies, flesh flies, and house flies can provide excellent evidence for forensic entomologists and are also essential to the fields of public health, medicine, and animal health. In all questions, the correct identification of fly species is an important initial step. The usual methods based on morphology or even molecular approaches can reach their limits here, especially when dealing with larger numbers of specimens. Since machine learning already plays a major role in many areas of daily life, such as education, business, industry, science, and medicine, applications for the classification of insects have been reported. Here, we applied the decision tree method with wing morphometric data to construct a model for discriminating flies of three families [Calliphoridae, Sarcophagidae, Muscidae] and seven species [Chrysomya megacephala (Fabricius), Chrysomya rufifacies (Macquart), Chrysomya (Ceylonomyia) nigripes Aubertin, Lucilia cuprina (Wiedemann), Hemipyrellia ligurriens (Wiedemann), Musca domestica Linneaus, and Parasarcophaga (Liosarcophaga) dux Thomson]. One hundred percent overall accuracy was obtained at a family level, followed by 83.33% at a species level. The results of this study suggest that non-experts might utilize this identification tool. However, more species and also samples per specimens should be studied to create a model that can be applied to the different fly species in Thailand.

Matched MeSH terms: Sarcophagidae*
Fulltext A new record of Iranihindia martellata (Senior-White,1924) (Diptera: Sarcophagidae) from peninsular Malaysia and female identification using both morphology and DNA-based approaches

Tan SH, Mohd Aris E, Kurahashi H, Mohamed Z

Trop Biomed, 2010 Aug;27(2):287-93.
PMID: 20962727

Iranihindia martellata (Senior-White, 1924) is recorded from peninsular Malaysia for the first time. Male and female specimens in the recent collections of forensically important sarcophagid flies were examined and identified based on morphology and DNA sequencing analysis. Male genitalia offer unambiguous species identification characteristics in the traditional taxonomy of flesh flies but the female flies are very similar to one another in general morphology. Female of I. martellata was determined by DNA sequencing (COI and COII) and PCR-RFLP (COI) analysis. Identified females were carefully examined and compared with the morphologically similar species, Liopygia ruficornis (Fabricius, 1794). Female genitalia are re-described and illustrated in this paper.

Matched MeSH terms: Sarcophagidae/anatomy & histology*; Sarcophagidae/classification*
Two new records of flies causing myiasis from Saudi Arabia with a survey of flies parasitizing goats and sheep in Jazan Region

Alahmed AM, Nasser MG, Sallam MF, Dawah H, Kheir S, AlAshaal SA

Trop Biomed, 2020 Jun 01;37(2):499-512.
PMID: 33612819

Despite the medical, veterinary and forensic relevance of myiasis-causing flies, knowledge of their diversity in Saudi Arabia is limited especially in the southern region. Therefore, a survey of myiasis-causing flies in the Jazan region was carried out using Red Top Fly Catcher traps baited with either decomposing beef liver or a lure composed primarily from fishmeal during the period April 2013-March 2014. Twelve known species were identified and recorded in this study, seven species of them belonging to Calliphoridae (Chrysomya, Lucilia, and Hemipyrella) and five species belonging to Sarcophagidae (Sarcophaga). Two of these species were recorded for the first time for Saudi Arabia, namely Hemipyrella pulchra (Wiedemann, 1830) and Sarcophaga (Liosarcophaga) exuberans Pandellé, 1896. Images of the species recorded are also provided for the first time. The results expand the knowledge of geographical distribution, fauna, and habitat of the myiasis-causing flies in Saudi Arabia. Biological information and world-wide geographical distribution of these species are included together with some taxonomic remarks.

Matched MeSH terms: Sarcophagidae/classification*
Random forests for predicting species identity of forensically important blow flies (Diptera: Calliphoridae) and flesh flies (Diptera: Sarcophagidae) using geometric morphometric data: Proof of concept

Khang TF, Mohd Puaad NAD, Teh SH, Mohamed Z

J Forensic Sci, 2021 May;66(3):960-970.
PMID: 33438785 DOI: 10.1111/1556-4029.14655

Wing shape variation has been shown to be useful for delineating forensically important fly species in two Diptera families: Calliphoridae and Sarcophagidae. Compared to DNA-based identification, the cost of geometric morphometric data acquisition and analysis is relatively much lower because the tools required are basic, and stable softwares are available. However, to date, an explicit demonstration of using wing geometric morphometric data for species identity prediction in these two families remains lacking. Here, geometric morphometric data from 19 homologous landmarks on the left wing of males from seven species of Calliphoridae (n = 55), and eight species of Sarcophagidae (n = 40) were obtained and processed using Generalized Procrustes Analysis. Allometric effect was removed by regressing centroid size (in log10 ) against the Procrustes coordinates. Subsequently, principal component analysis of the allometry-adjusted Procrustes variables was done, with the first 15 principal components used to train a random forests model for species prediction. Using a real test sample consisting of 33 male fly specimens collected around a human corpse at a crime scene, the estimated percentage of concordance between species identities predicted using the random forests model and those inferred using DNA-based identification was about 80.6% (approximate 95% confidence interval = [68.9%, 92.2%]). In contrast, baseline concordance using naive majority class prediction was 36.4%. The results provide proof of concept that geometric morphometric data has good potential to complement morphological and DNA-based identification of blow flies and flesh flies in forensic work.

Matched MeSH terms: Sarcophagidae/anatomy & histology*
Larval growth of Liosarcophaga dux Thompson (Diptera: Sarcophagidae) under uncontrolled indoor temperatures in Malaysia

Kumara TK, Hassan AA, Salmah MR, Bhupinder S

Southeast Asian J Trop Med Public Health, 2013 Mar;44(2):182-7.
PMID: 23691627

The larval growth of Liosarcophaga dux Thompson (Diptera: Sarcophagidae) was studied under varying indoor room temperatures in Malaysia. Five replicates were established. The immature growth of this species from first instar until adult emergence was 307.0+/-3.0 hours. The mean larval length measured for second instar, third instar, post-feeding stage and puparia were 6.5+/-0.5 mm (n=10), 11.8+/-3.7 mm (n=31), 12.7+/-0.8 mm (n=16), and 9.5+/-0.5 mm (n=15), respectively.

Matched MeSH terms: Sarcophagidae/growth & development*
Fulltext Review of forensically important entomological specimens in the period of 1972 - 2002

Lee HL, Krishnasamy M, Abdullah AG, Jeffery J

Trop Biomed, 2004 Dec;21(2):69-75.
PMID: 16493401

Forensic entomological specimens received by the Unit of Medical Entomology, IMR., from hospitals and the police in Malaysia in the last 3 decades (1972 - 2002) are reviewed. A total of 448 specimens were received. From these, 538 identifications were made with the following results: Eighteen species of cyclorrphaga flies were identified consisting of Chrysomya megacephala (Fabricius) 215 cases (47.99%), Ch. rufifacies (Masquart) 132 (29.46%), Ch. villeneuvi Patton 10 (2.23%), Ch. nigripes Aubertin 7 (1.56%), Ch. bezziana Villeneuve 4 (0.89%), Ch. pinguis (Walker) 1 (0.22%), Chrysomya sp. 47 (10.49%), Sarcophaga sp. 28 (6.25%), Lucilia sp. 21 (4.69%), Hermetia sp. 15 (3.35%), He. illucens (Linnaeus) 1 (0.22%), Hemipyrellia ligurriens (Wiedemann) 3 (0.67%), Hemipyrellia sp. 2 (0.45%), Ophyra spinigera 1 (0.22%), Ophyra sp. 6 (1.34%), Calliphora sp. 24 (5.36%), Synthesiomyia nudiseta (Wulp) 1 (0.22%) and Eristalis sp. 1 (0.22%). Other non - fly insect specimens are Pthirus pubis (Linnaeus) (Pubic louse) 2 (0.45%) and Coleoptera (Beetles) 1 (0.22%). Ch. megacephala and Ch. rufifacies were the commonest species found in cadavers from different ecological habitats. Sy. nudiseta is an uncommon species, thus far found only on cadavers from indoors. Sy. nudiseta is reported for the second time in Peninsular Malaysia. A total of 329 cases (73.44%) had a single fly infestation, 109 cases (24.33%) had double fly infestation and 10 cases (2.23%) had triple fly infestation. Five cases (1.12%) had eggs and 3 cases (0.67%) had larval stages that were not identifiable. No arthropods were retrieved from cadavers in 8 cases (1.79%). In conclusion, although large number of fly species were found on human cadavers, the predominant species are still those of Chrysomya.

Matched MeSH terms: Sarcophagidae
Fulltext Flies specimens collected from Agricultural Park, Teluk Cempedak and Bukit Pelindung in Kuantan, Pahang

Heo, Chong Chin, Mohamad Abdullah Marwi, Jeffery, John, Ismarulyusda Ishak, Baharudin Omar

Jurnal Sains Kesihatan Malaysia, 2008;6(2):93-99.
MyJurnal

This study was carried out in Agricultural Park, Teluk Cempedak and Bukit Pelindung at Kuantan, Pahang in October 2007. These three areas were different in ecological characteristic, Agricultural Park is a lowland region in Kuantan rural area, Teluk Cempedak is Kuantan’s most famous beach, and Bukit Pelindung is a reserved rainforest which is 200 meters from the sea level. Fly specimens were collected using four different kinds of baits: dry prawn, salted fish, pork and mango. Each of these baits was placed in a plastic container and exposed for one hour to attract flies. Within 5 minutes, flies started swarming around the baits. The flies were more attracted to the pork and salted fish compared to the other two baits. Fifty one flies, one moth (Lepidoptera) and one wasp (Hymenoptera) were collected. In Agricultural Park, two Lucilia cuprina, one Chrysomya megacephala and one Sarcophaga sp. were collected. For Teluk Cempedak beach, there were two Sarcophagids, 31 Chrysomya megacephala, five Musca domestica, one Lucilia cuprina and one moth were caught. Flies collected from Bukit Pelindung included five C.megacephala, two Sarcophagids, one Musca domestica and one wasp. Most C.megacephala were attracted to the pork and salted fish.

Matched MeSH terms: Sarcophagidae
Opportunistic insects associated with pig carrions in Malaysia

Heo CC, Kurahashi H, Mohamed Abdullah Marwi, Jeffery J, Baharudin Omar

Sains Malaysiana, 2011;40:1179-1186.

Flies from the family Calliphoridae, Sarcophagidae and Muscidae are usually found on human cadavers or animal carcasses. However, there are many other families of Diptera and Coleoptera that were found associated with animal carcasses, which have not been reported in Malaysia. In this paper, we report dipterans from the family Micropezidae: Mimegralla albimana Doleschall, 1856, Neriidae: Telostylinus lineolatus (Wiedemann 1830); Sepsidae: Allosepsis indica (Wiedemann 1824), Ulidiidae: Physiphora sp. and a beetle (Coleoptera: Hydrophilidae: Sphaeridium sp.) as opportunist species feeding on oozing fluid during the decomposition process. They did not oviposit on the pig carcasses, therefore, their role in estimation of time of death is of little importance. However, they could provide clues such as locality and types of habitats of the crime scene.

Matched MeSH terms: Sarcophagidae
Fulltext Scuttle Flies (Diptera: Phoridae) Inhabiting Rabbit Carcasses Confined to Plastic Waste Bins in Malaysia Include New Records and an Undescribed Species

Zuha RM, Huong-Wen S, Disney RH, Omar B

Trop Life Sci Res, 2017 Jan;28(1):131-143.
PMID: 28228921 MyJurnal DOI: 10.21315/tlsr2017.28.1.9

Scuttle flies (Diptera: Phoridae) are small-sized insects of forensic importance. They are well known for diversified species and habitats, but in the context of forensic entomology, scuttle flies' inhabitance of corpses remains inadequately explored. With recent reports indicating the existence of more scuttle fly species possibly inhabiting these environments, a decomposition study using animal carcasses in enclosed environments was conducted. The aim was to record the occurrence of scuttle flies on rabbit carcasses placed in sealed plastic waste bins for a 40-day period. The study was conducted as two replicates in Bangi, Selangor. Sampling was carried out at different time intervals inside a modified mosquito net as a trap. Inside the trap, adult scuttle flies were aspirated and preserved in 70% ethanol. The fly larvae and pupae were reared until their adult stage to facilitate identification. From this study, six scuttle fly species were collected, i.e., Dahliphora sigmoides (Schmitz) ♂, Gymnoptera simplex (Brues) ♀, Megaselia scalaris (Loew) ♂♀, Puliciphora borinquenensis (Wheeler) ♂, Puliciphora obtecta Meijere ♀ and Spiniphora sp. ♀. Both D. sigmoides and P. obtecta were newly recorded in Malaysia, whilst the Spiniphora sp. was considered an unknown species until it was linked to its male counterpart. The sealed waste bins were found to be accessible for the scuttle flies with delayed arrival (day 4-5). Megaselia scalaris was the primary scuttle fly species attracted to the carcass, and its occurrence could be observed between days 4-7 (replicate 1) and days 5-33 (replicate 2). This study also revealed Sarcophaga spp. (Diptera: Sarcophagidae) as the earliest species to colonize the remains and the longest to inhabit them (days 2-40). The larvae of Hermetia illucens (Linneaus) (Diptera: Stratiomyidae) and Fannia sp. (Diptera: Fanniidae) were found on the carcasses during the mid-advanced decay period. These findings expand the knowledge on the diversity of forensically important scuttle flies and coexisting dipterans in enclosed environments in Malaysia.

Matched MeSH terms: Sarcophagidae
Fulltext Determination of antibacterial activity and minimum inhibitory concentration of larval extract of fly via resazurin-based turbidometric assay

Teh CH, Nazni WA, Nurulhusna AH, Norazah A, Lee HL

BMC Microbiol, 2017 Feb 16;17(1):36.
PMID: 28209130 DOI: 10.1186/s12866-017-0936-3

BACKGROUND: Antimicrobial resistance is currently a major global issue. As the rate of emergence of antimicrobial resistance has superseded the rate of discovery and introduction of new effective drugs, the medical arsenal now is experiencing shortage of effective drugs to combat diseases, particularly against diseases caused by the dreadful multidrug-resistant strains, such as the methicillin-resistant Staphylococcus aureus (MRSA). The ability of fly larvae to thrive in septic habitats has prompted us to determine the antibacterial activity and minimum inhibitory concentrations (MICs) of larval extract of flies, namely Lucilia cuprina, Sarcophaga peregrina and Musca domestica against 4 pathogenic bacteria [Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Pseudomonas aeruginosa and Escherichia coli] via a simple and sensitive antibacterial assay, resazurin-based turbidometric (TB) assay as well as to demonstrate the preliminary chemical profile of larval extracts using gas chromatography-mass spectrophotometry (GC-MS).
RESULTS: The resazurin-based TB assay demonstrated that the L. cuprina larval extract was inhibitory against all tested bacteria, whilst the larval extract of S. peregrina and M. domestica were only inhibitory against the MRSA, with a MIC of 100 mg ml(-1). Subsequent sub-culture of aliquots revealed that the larval extract of L. cuprina was bactericidal against MRSA whilst the larval extracts of S. peregrina and M. domestica were bacteriostatic against MRSA. The GC-MS analysis had quantitatively identified 20 organic compounds (fatty acids or their derivatives, aromatic acid esters, glycosides and phenol) from the larval extract of L. cuprina; and 5 fatty acid derivatives with known antimicrobial activities from S. peregrina and M. domestica.
CONCLUSION: The resazurin-based turbidometric assay is a simple, reliable and feasible screening assay which evidently demonstrated the antibacterial activity of all fly larval extracts, primarily against the MRSA. The larval extract of L. cuprina exerted a broad spectrum antibacterial activity against all tested bacteria. The present study revealed probable development and use of novel and effective natural disinfectant(s) and antibacterial agent(s) from flies and efforts to screen more fly species for antibacterial activity using resazurin-based TB assay should be undertaken for initial screening for subsequent discovery and isolation of potential novel antimicrobial substances, particularly against the multi-drug resistant strains.

Matched MeSH terms: Sarcophagidae/chemistry

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