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Report on the occurrence of synanthropic derived form of Chrysomyamegacephala (Diptera: Calliphoridae) from Royapuram fishing harbour, Chennai, Tamil Nadu, India
Mr Paulchamy Ramaraj; Dr Chellappa Selvakumar; Mr Arumugam Ganesh; Dr Sundaram Janarthanan
Biodiversity Data Journal , iss. p.

AbstractEdit

AbstractThe occurrence of dipteran fly, Chrysomyamegacephala (Fabricius, 1794) is reported for the first time from Royapuram fishing harbour (Chennai), Tamil Nadu, South East India. The fully grown third instar larvae of Chrysomyamegacephala were collected from decaying fishes near Royapuram fishing harbour. This site is found to be the regular breeding site for Chrysomyamegacephala. Larvae were reared under laboratory condition and freshly emerged adult flies from pupae were collected and identified by morphological features and molecular tools. Molecular identification through generation of DNA barcoding using mitochondrial COI gene of Chrysomyamegacephala is appended.

IntroductionEdit

Calliphoridae is a cosmopolitan group of calyptrate flies comprising nearly 1500 recognized species worldwide ([1]). The Indian representatives of this family belong to 63 species under 18 genera ([2]; [3], [4]). Chrysomya is one of the most significant genera of Calliphoridae with great medical, veterinary and forensic importance. Ten species of the genus Chrysomya have been recorded in India ([5]).

Chrysomyamegacephala (Fabricius, 1794) is commonly found in cadavers in many parts of the world ([6], [7], [8]), and is used in forensic investigations to determine post-mortem intervals ([9], [10], [11], [12], [13]). It is a tremendously abundant species, dispersed rapidly throughout the Oriental, Australasian, Afrotropical, Neotropical, Nearctic and Palearctic zoogeographical regions, ([14], [15]), and used as fish bait in northern and south-west India ([16], [17]). It is known to breed in human faeces, decomposed meat and fish ([18], [19]) as well as discarded organic materials. This dipteran fly, therfore, serves as a potential vector of many diseases due to its close association with human dwellings. In general, Chrysomyamegacephala is considered important in medical, veterinary and forensic sciences ([20], [21], [13], [14], [22]).

Presently, three forms of Chrysomyamegacephala are recognized, namely, the normal form (nf), the synanthropic derived form (sdf) and the recently reported feral derived form (fdf) ([23]). The normal form is confined to forests of South Pacific Islands (from Bismarck Archipelago to Western Samoa) while synanthropic derived form has spread around the world from Papua New Guinea. The recently reported feral derived form (fdf) in the forests of the Himalaya, India is a morphological intermediate between normal and synanthropic derived forms ([23]). Synanthropic derived form of Chrysomyamegacephala is common around human dwellings enabling direct contact with humans. It occurs on dead fish, sweets, carcasses, human excrement and fruits. The adult flies are vectors of a few infectious diseases of the digestive tract and reported to carry Morganella sp. (Enterobacteriales: Enterobacteriaceae), which causes summer diarrhoea ([24]). It is also associated with many microbial pathogens including polio virus. Besides, this fly has been reported as vectors of enteric pathogens in malnourished individuals living under unsanitary conditions ([9]). The larvae of Chrysomyamegacephala are parasitic on semi-dried fish, causing a major problem in the fish industry of Southeast Asia ([25], [17]). It is a secondary myiasis producer in man and in domestic animals ([24]).

However, the occurrence of the synanthropic derived form of Chrysomyamegacephala has not been documented in the State of Tamil Nadu, India. In this context, the present study reports for the first time the synanthropic derived form of Chrysomyamegacephala from Royapuram fishing harbour, Chennai, Tamil Nadu, South East India and provides key characters based on morphological features and molecular analysis.

Materials and methodsEdit

Collection, rearing and morphological identification

A colony of Chrysomyamegacephala was established from numerous second and third instar larvae collected from decaying fishes of Royapuram fishing harbour in North Chennai, Tamil Nadu, South India (Fig. 1). The colony of Chrysomyamegacephala used in this study was successfully cultured in the insectarium at the Unit of Entomology, Department of Zoology, University of Madras. Colonies were maintained at 28 °C and R.H. of 80%. Freshly emerged adults from pupae were morphologically identified using the keys provided by [26]. Voucher specimens were deposited at Zoological Survey of India, Southern Regional Centre, Chennai, Tamil Nadu, India (I/D 28: 3 male and I/D 29: 3 female).

Morphological descriptionEdit

The procedure adopted to identify the morphological features and terminology used in the description were based on the previous reports of [27], [28], [24].

Molecular identificationEdit

Genomic DNA was extracted from a single morphologically identified adult male and female species of Chrysomyamegacephala (sdf) after removing the gut region as per the standard phenol/chloroform extraction protocol ([29]).The extracted DNA was quantified using spectrophotometer and diluted to 50ng/µl prior to PCR amplification. The mitochondrial cytochrome-c oxidase subunit I (COI) gene was amplified using the primer pair LCO1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO2198 (5’ TAAACTTCAGGGTGACCAAAAAATCA-3’) under standard PCR conditions ([30]). The amplified PCR product was purified from agarose gel using PCR product purification kit (Qiagen). The purified PCR products were used for sequencing with Big Dye on ABI 3730 automated DNA sequencer. BLAST analysis was performed using these sequences to identify matches from the database.

Taxon treatmentsEdit

ChrysomyamegacephalaEdit

Fabricius, 1794

AB910389

AB910390

SPLID033-14

SPLID013-13

MaterialsEdit

  1. Type status:Other material. Occurrence: catalogNumber: I/D 28: 3 male and I/D 29: 3 female; recordedBy: Dr. S. Janarthanan; individualCount: 25; sex: 12 male, 13 female; disposition: Zoological Survey of India, Southern Regional Centre, Chennai, Tamil Nadu, India; Taxon: kingdom: Animalia; phylum: Arthropoda; class: Insecta; order: Diptera; family: Calliforidae; genus: Chrysomya; specificEpithet: megacephala; taxonRank: species; vernacularName: Oriental latrine fly; taxonomicStatus: accepted; Location: country: India; stateProvince: Tamil Nadu; municipality: Chennai; verbatimLocality: Royapuram fishing horbour; verbatimElevation: 2 m; verbatimLatitude: 13°07'44.73 N; verbatimLongitude: 80°17'52.70 E; Identification: identifiedBy: P. Ramaraj & C. Selvakumar; dateIdentified: 29 Jan 2014; identificationReferences: Senior-White et al., 1940; identificationRemarks: Eye facets of upper two-thirds greately enlarged and sharply demarcated from small facets of lower third; Event: samplingProtocol: Hand picking; eventDate: 2013-12-22; Record Level: institutionCode: University of Madras, Chennai

DiagnosisEdit

Larva: Fully mature third instar muscoid-shaped (11 mm length), composed of 12 segments with pointed anterior and blunt posterior end (Fig. 2a). Cephaloskeleton present in anterior end; dorsal cornu longer than ventral cornua; accessory sclerite, parastomal sclerite present; anterodorsal process apparent (Fig. 2b). Anterior spiracle possessed eight to 12 marginal branches (Fig. 2c). Dorsal cuticular spines between the prothorax and mesothorax arranged single, each having one to three dark pointed tips (Fig. 2d). Inner dorsal tubercle moderate between middle dorsal tubercle and outer dorsal tubercle, with latter tubercle largest; a pair of the posterior spiracles clearly visible at the caudal end (Fig. 2e). Peritreme incomplete and heavily pigmented as a dark structure encircling three relatively straight spiracular openings (slits), with middle slits appearing slightly bent inward (Fig. 2f).

PupaEdit

Larger in size than larvae; cylindrical in shape (Fig. 3a). A pair of anterior spiracles located laterally on the prothorax at the anterior tip (Fig. 3b). A pair of posterior spiracles located prominently on the contracted surface of caudal region. Each spiracle more or less an oval shaped peritreme, encircling three straight spiracular slits (Fig. 3c).

AdultEdit

Male (Fig. 4a): Body length 10-11 mm.

Head: Eye facets of upper two-thirds greatly enlarged and sharply demarked from small facets of lower third (Fig. 4c). Antenna short, postpedicel four times length of pedicel, pedicel dark brown, third brownish; arista dark brown, long plumose; palpi yellowish and slender (Fig. 5a). Thorax: Prostigmatic bristles present; anterior and posterior thoracic spiracles dark brown. Wings: Stem-vein with a row of setulae on upper posterior side; upper surface of lower lobe of squama hairy; upper squama white, partly dark margined; lower squama brown (Fig. 5b). Abdomen: Genital tergites bluish with black hairs; inner forceps elongated, outer forceps reduced (Fig. 5c); anterior paramere wide, slightly curved anteriorly; posterior paramere bifurcated at end; acrophallus wide at end and with a wide stakled projection posteriorly (Fig. 5d).

Female: (Fig. 4b). Body length 10-11 mm.

Head: Eyes separated by one-quarter total width of head; facets uniformly small; frons clearly separated; widest part of frons more than width of ocellar triangle; frontal reddish to black, with small hairs on the upper part; frontal bristles short and weak; parafrontalia slightly narrower than width of frons, covered with golden tomentum, but appearing black towards vertex in certain lights; parafacilia yellowish brown with silvery pollen and white hairs (Fig. 4d).

Molecular identification and DNA barcodingEdit

The 651 bp mitochondrial cytochrome c oxidase subunit I (COI) sequences of Chrysomyamegacephala (sdf) were 100% identical with available NCBI database sequences of Chrysomyamegacephala using BLAST analysis. These mtCOI gene sequences of Chrysomyamegacephala were submitted to NCBI database (Accession No. [AB910389 AB910389]-male; [AB910390 AB910390]-female) and DNA barcodes were generated for both the sexes based on their COI sequences using Barcode of Life Database (BOLD system; Process ID SPLID013-13 and SPLID033-14).

DistributionEdit

South India – Tamil Nadu (Chennai); Kerala (Calicut); North India - West Bengal and rapidly throughout the continent.

EcologyEdit

Life cycle: Second and third instar larvae of Chrysomyamegacephala (sdf) were collected from decaying fishes of Royapuram fishing harbour of North Chennai, Tamil Nadu, South India. Adults emerged in the rearing chamber 7-9 d after pupation. Mating began 2 d after emergence and oviposition occurred at 3-4 d of age and laid egg mass on pieces of spoiled chicken in a squat cup. Each egg mass contained 200-300 eggs. Eggs held at 28 °C hatched on 1 d. Second instars emerged 2 d after hatching, becoming 3rd instar 2 d later, and then pupation occurred 4-6 d.

DiscussionEdit

This study reports for the first time the occurrence of Chrysomyamegacephala (sdf) in Tamil Nadu, India and provides, therefore, key identification characters of this blowfly based on distinct morphological features of larvae, pupae and adult males and females as well as molecular barcode analysis in the adults of both sexes. Chrysomyamegacephala also known as the oriental latrine fly ([31]), and it is considered one of the most dangerous dipteran vector ([32]) imposing a heavy potential hygienic risk ([33]). This fly has been expanding its range throughout the world. In India, this species has been reported from north and south-west regions ([16], [17], [2], [23], [5]). The present study is a report on the occurrence of Chrysomyamegacephala for the first time in south-east India (coastal regions of Tamil Nadu, especially Royapuram coast) in considerable abundance on fish wastes.

Supplementary MaterialEdit

XML Treatment for ChrysomyamegacephalaEdit

File:BDJ.2.e1111-treatment1.xml

AcknowledgementsEdit

The authors thank the financial assistance received from UGC-SAP. C. Selvakumar thanks University Grants Commission, NewDelhi for the award of Dr. D. S. Kothari Post Doctoral Fellowship [No.F.4-2/2006(BSR) /13-670/2012 (BSR)].

Author contributionsEdit

All authors are equally contributed

ReferencesEdit

  1. ↑ De Carvalho and Mello-Patiu 2008 Cite error: Invalid <ref> tag; name "B670022" defined multiple times with different content
  2. ↑ 2.0 2.1 Nandi 2004 Cite error: Invalid <ref> tag; name "B670302" defined multiple times with different content
  3. ↑ Singh and Sidhu 2004 Cite error: Invalid <ref> tag; name "B670332" defined multiple times with different content
  4. ↑ Singh and Sidhu 2007 Cite error: Invalid <ref> tag; name "B670342" defined multiple times with different content
  5. ↑ 5.0 5.1 Bharti 2011 Cite error: Invalid <ref> tag; name "B669964" defined multiple times with different content
  6. ↑ Gruner et al. 2007 Cite error: Invalid <ref> tag; name "B670224" defined multiple times with different content
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  13. ↑ 13.0 13.1 Centeno et al. 2002 Cite error: Invalid <ref> tag; name "B670012" defined multiple times with different content
  14. ↑ 14.0 14.1 Smith 1986 Cite error: Invalid <ref> tag; name "B670382" defined multiple times with different content
  15. ↑ Castro and García 2009 Cite error: Invalid <ref> tag; name "B669974" defined multiple times with different content
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  17. ↑ 17.0 17.1 17.2 Wall et al. 2001 Cite error: Invalid <ref> tag; name "B670467" defined multiple times with different content
  18. ↑ Laurence 1981 Cite error: Invalid <ref> tag; name "B670234" defined multiple times with different content
  19. ↑ Laurence 1986 Cite error: Invalid <ref> tag; name "B670244" defined multiple times with different content
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  21. ↑ De Souza and Linhares 1997 Cite error: Invalid <ref> tag; name "B670046" defined multiple times with different content
  22. ↑ Wells and Kurahashi 1994 Cite error: Invalid <ref> tag; name "B670507" defined multiple times with different content
  23. ↑ 23.0 23.1 23.2 Bharti and Kurahashi 2009 Cite error: Invalid <ref> tag; name "B669954" defined multiple times with different content
  24. ↑ 24.0 24.1 24.2 Sinha and Nandi 2004 Cite error: Invalid <ref> tag; name "B670352" defined multiple times with different content
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  26. ↑ Senior-White et al. 1940 Cite error: Invalid <ref> tag; name "B670322" defined multiple times with different content
  27. ↑ Sukontason et al. 2008 Cite error: Invalid <ref> tag; name "B716584" defined multiple times with different content
  28. ↑ Siriwattanarungsee et al. 2005 Cite error: Invalid <ref> tag; name "B716611" defined multiple times with different content
  29. ↑ Murry and Thompson 1980 Cite error: Invalid <ref> tag; name "B670292" defined multiple times with different content
  30. ↑ Folmer et al. 1994 Cite error: Invalid <ref> tag; name "B670095" defined multiple times with different content
  31. ↑ Zumpt 1965 Cite error: Invalid <ref> tag; name "B670691" defined multiple times with different content
  32. ↑ Wells 1991 Cite error: Invalid <ref> tag; name "B670517" defined multiple times with different content
  33. ↑ Maldonado and Centeno 2003 Cite error: Invalid <ref> tag; name "B670254" defined multiple times with different content

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