Journal of Threatened Taxa | www.threatenedtaxa.org | 26 January 2022 | 14(1): 20503–20510

 

ISSN 0974-7907 (Online) | ISSN 0974-7893 (Print) 

https://doi.org/10.11609/jott.7638.14.1.20503-20510

#7638 | Received 31 August 2021 | Final received 26 November 2021 | Finally accepted 03 January 2022

 

 

Morphological characterization and mt DNA barcode of a tiger moth species, Asota ficus (Fabricius, 1775) (Lepidoptera: Noctuoidea: Erebidae: Aganainae) from India

 

Aparna Sureshchandra Kalawate 1 , K.P. Dinesh 2  & A. Shabnam 3

 

1,2,3 Zoological Survey of India, Western Regional Centre, Vidya Nagar, Sector-29, P.C.N.T. (PO), Rawet Road, Akurdi, Pune, Maharashtra 411044, India

1 devarpanento@gmail.com (corresponding author), 2 kpdinesh.zsi@gmail.com, 3 shabnamansari9113@gmail.com

 

 

 

Editor: Mandar Paingankar, Government Science College Gadchiroli, Maharashtra, India. Date of publication: 26 January 2022 (online & print)

 

Citation: Kalawate, A.S., K.P. Dinesh & A. Shabnam (2022). Morphological characterization and mt DNA barcode of a tiger moth species, Asota ficus (Fabricius, 1775) (Lepidoptera: Noctuoidea: Erebidae: Aganainae) from India. Journal of Threatened Taxa 14(1): 20503–20510. https://doi.org/10.11609/jott.7638.14.1.20503-20510

 

Copyright: © Kalawate et al. 2022. Creative Commons Attribution 4.0 International License.  JoTT allows unrestricted use, reproduction, and distribution of this article in any medium by providing adequate credit to the author(s) and the source of publication.

 

Funding: The work is based on the annual research programme of Zoological Survey of India, WRC, Pune ( Ministry of Environment

& Forests, Govt. of India).

 

Competing interests: The authors declare no competing interests.

 

Acknowledgements: Authors are grateful to the director, Zoological Survey of India, Kolkata and the officer-in-charge, Zoological Survey of India, Western Regional Centre, Pune for encouragement and research facilities. We are grateful to Dr. R.M. Sharma, retired scientist and the officer-in-charge, Zoological Survey of India, Western Regional Centre, Pune for critically going through the manuscript. Due acknowledgements to the survey team members of Zoological Survey of India, Western Regional Centre, Pune for collection efforts. The help of Ms. Mehrun Raje in the wet lab studies are acknowledged. Authors are thankful to the anonymous reviewers and the subject editor for their valuable suggestions and constructive criticism on the earlier version of the manuscript.

 

 

 

Abstract: The members of the genus Asota are widely distributed from Africa, India, Sri Lanka, Myanmar, and Malayan regions to the Australian region containing 55 described species. Asota ficus (Fabricius, 1775) is one among the nine species of the genus described from India having a wide range of distribution. The present study includes the first mitochondrial DNA barcode generated from India for A. ficus with a valid voucher describing external morphological characters together with the male and female genitalia. Discussions pertain to the utility of DNA barcodes for studies on moths in India with a comment on the identity of other sequences showing shallow genetic divergence with our sequences.

 

Keywords:  Arctiinae, Ficus, genitalia study, Hypsa, Lepidopterism, Maharashtra, Mitragyna, molecular study, mt COI, Ricinus.

 

 

 

The subfamily Aganainae Boisduval, 1833 was earlier considered as family Aganaidae or Hypsidae (Inoue et al 1982). Later studies considered it as subfamily Hypsinae of Arctiidae (Seitz 1914; Daniel 1943) or subfamily Aganainae of Noctuidae (Holloway 1988; Scoble 1992; Kitching & Rawlins 1998). Until molecular studies, the familial position was unstable, later on phylogenetic studies placed the subfamily Aganainae under the family Erebidae (Fibiger & Lafontaine 2005; Zahiri et al. 2012). Aganainae includes 109 species of 11 genera worldwide (Zahiri et al. 2012; Bayarsaikhan et al. 2016).

Many Aganainae moths are large, brightly coloured, aposematic, with bare lower frons and long upturned labial palps having long and slender third segment; vein M2 in forewing arises closer to the origin of M3 than M1, in the lower part of the discal cell; Cu appearing four-branched; vein M2 in the hindwing is present so Cu appears four-branched (Holloway 1988; Zahiri et al. 2012). The larvae have single subventral seta on the mesothoracic and metathoracic segments. The subfamily exhibits a sister relationship with Arctiinae with a strongly supported pairing (Zahiri et al 2011).

Moths from this subfamily are pests on plant species of Apocynaceae, Asclepiadaceae, Moraceae (Holloway 1988; Common 1990; Bayarsaikhan et al. 2016), and lactiferous families that contain cardenolides (Bayarsaikhan et al. 2016). They feed on poisonous plants, and hence are often aposematic day flyers (Kitching & Rawlins 1998; Bayarsaikhan et al. 2016).

The genus Asota Hübner, [1819] was erected by Jacob Hubner in 1819 considering Phalaena javana (Cramer, [1780]) from Java as type species. So far, 55 species are known from this genus including nine from India. The Asota species reported from India are: caricae (Fabricius, 1775); plana (Walker, 1854); canaraica (Moore, 1878); egens (Walker, 1854); ficus (Fabricius, 1775); heliconia (Linnaeus, 1758); paphos (Fabricius, 1787); producta (Butler, 1875); sericea (Moore, 1878). A. ficus was placed under the genus Hypsa as Hypsa ficus by Hampson (1892) under the family Hypsidae: section-II. Hampson (1892) divided the genus Hypsa under two sections on the basis of structure of antennae. In Section-I the antennae of males are fasciculated with short cilia. The fasciculated male antennae, long cilia and the long 3rd segment of palpi forms the section-II. Caterpillar of A. ficus is recorded feeding mainly on castor and ficus.

The genus Asota is responsible for Lepidopterism, a disease caused by the adult or the caterpillar of moths or butterflies (Wills et al. 2016).  In Kerala India, it was reportedly caused by the tiger moth A. caricae (Anonymous 2016). The fever caused by Lepidopterism mimics the symptoms of the mosquito borne infectious diseases like chikungunya and dengue. The adult moths, while emerging from the pupae, extricate the scales on their body and secretes fluids (Anonymous 2016)  which lead to the high fever either when in contact with the human skin or due to inhalation. As per Wills et al. (2016), allergic reactions are due to the presence of poisonous chemicals like histamines, imidazole and peptides.

DNA barcoding is a quick and reliable nucleotide-based identification technique across the animal kingdom, founded on the mitochondrial Cytochrome oxidase I gene (mt COI) by Hebert’s group in 2003. The ability of COI sequences to discriminate closely allied species based on restricted intraspecific mitochondrial DNA divergence and utilizing it as an aid to resolve the alpha diversity of species in diverse taxonomic groups including Lepidoptera has been validated (Hebert et al. 2003b). These species-specific signatures, identified as DNA barcodes help to delimit the problematic taxa (Hebert et al. 2003a) also in cases where identification is not possible with the traditional taxonomic techniques alone. DNA barcode not only provides a boon to taxonomic research but also serves as a form of comprehensive, widely accessible system for identification and validation of species. Hence, in the present study an attempt has been made to develop a DNA barcode for the species A. ficus from Maharashtra along with its morphological description (adult together with external genitalia); the utility of mt DNA barcodes in the Indian moth studies are discussed.

 

Materials and Methods

Moth specimens were collected using a light trap having mercury vapour lamp as a light source of 160 W. It was hung in the middle of the white sheet installed in the field during the night. Moth specimens that were captured were euthanized by ethyl acetate vapours. Then they were transported to the laboratory in insect packets (made of butter paper) for further analysis.

In the laboratory, the specimens were stretched, pinned and stored in entomological boxes filled with preservatives. For morphological studies the specimens were studied under Leica EZ4E stereomicroscope. The map of the collection locality was prepared using open free QGIS software. The details of the collection locality are given under the material examined and is also shown in Figure 1. Identification of the specimens was done as per Hampson (1892). Male and female genitalia were studied following Robinson (1976). The identified specimens are deposited at the National Zoological Collections of the Zoological Survey of India, Western Regional Centre, Pune, Maharashtra, India (ZSI/WRC).

DNA extraction was performed using DNeasy blood and tissue kit (Qiagen) using leg and abdomen of a dried specimen. DNA quantitation was performed by HS dsDNA assay kit on Qubit 2.0 fluorometer. Mitochondrial COI (mt COI) gene was amplified using universal primer pair, LCO1490 and HCO2198 (Folmer et al. 1994) in 25 µL reaction volume constituted by 12.5 µL of Master Mix (Promega), 10 pmol of each forward and reverse primer, 50 ng of template DNA along with Nuclease free water up to Q.S. Thermal cycling profile performed as per Kalawate et al. (2020a). Amplification of the desired gene was confirmed by gel electrophoresis stained by SYBR safe DNA gel stain (Invitrogen), visualized under UV by gel documentation system. Purification of the amplified product was done by Invitrogen’s Pure Link PCR Purification Kit. The purified PCR product was sequenced bi-directionally by Sanger’s method on ABI 377 (Applied Biosciences) sequencer.

Both the forward and reverse sequences generated in the current studies were verified manually for corrections. Initially 838 mt COI gene sequences available for the genus Asota were downloaded from the GenBank and were aligned using MEGA 5.2 software (Tamura et al 2011). MEGA 5.2 (Tamura et al. 2013) was used for calculating uncorrected pairwise genetic distances. Initial tree was built (using MEGA 5.2) including all reported species with molecular data for the genus Asota, comprising 235 sequences excluding identical sequences from the same locality for a single species/subspecies. Since mt COI is not a good candidate gene for phylogenetic studies (Cameron et al. 2004; Lafontaine & Schmidt 2010) and our initial single gene phylogenetic tree ended up in polytomies without proper phylogenetic relationships, we considered presenting the phylogenetic tree comprising all the sequences of A. ficus available on the GenBank with the sequences generated by us and the probable sister species A. speciosa treating species Neochera inops as an outgroup. The phylogenetic inferences drawn are only to show the monophyly of all the sequences of A. ficus. Maximum likelihood tree was generated using RaxML (Silvestro & Michalak 2012) with thorough bootstrap of 1,000 replicates under the GTR+GAMMA+I model and the final consensus tree was visualized by Fig Tree v1.4.0. Sequences generated in the studies are submitted to the GenBank (OL630456.1 & OL630457.1).

 

Result and Discussions

Taxonomic account

Superfamily Noctuoidea Latreille, 1809

Family Erebidae Leach, [1815]

Subfamily Aganainae Boisduval, 1833

Genus Asota Hübner, [1819]

Asota Hübner, [1819], Verz. bek. Schmett. (11): 164.

Type Species: Phalaena javana (Cramer, [1780])

 

Asota ficus (Fabricius, 1775)

Noctua ficus Fabricius,1775, Syst. Ent.: 595.

Lacides ficus, Moore,188, Lep. Ceylon, 2(1): 53, pl. 100, f. 2.

Hypsa ficus, Hampson,1892, Fauna Brit. India, Moths, 1: 504.

Type Locality. India.

Material examined/source: 01 male, Saptashringigadh, Nashik, Maharashtra, India (20.23N, 73.54E; 1,000 m), 06 November 2016, coll. A.S. Kalawate (ZSI/WRC/L-1482); 01 female, Ambegaon, Pune, Maharashtra, India (19.13N, 73.73E; 730 m), 23 June 2017, coll. A.S. Kalawate & party (ZSI/WRC/L-1780); 02 male, Bhaskaracharya Forest Rest house, Gautala, Jalgaon, Maharashtra, India (20.34N, 75.14E; 711 m), 27 September 2019, coll. P.S. Bhatnagar & party (ZSI/WRC/L-2069).

Morphological description: Adult (Image 1A,B). Wing expanse: 55 mm in male and 63 mm in female. Antennae of male fasciculated, cilia long; 3rd joint of palpi long, grey in colour, tipped with black. Head, thorax and abdomen orange-yellow; tegulae with yellow base and a black spot. Abdomen with series of black spots. Orange basal patch on forewing extending along costa and in cell to two-third length of cell, an orange spot encircled with black on the costa, and streaks in cell and on inner margin, two black spots on costa and in cell, one on inner margin, and two lines across interno-median interspace; rest of the wing olive-brown, the veins are striped with yellow. Hind wing bright orange-yellow; black spot at end of cell and series of irregular sized and placed black spots at submarginal area. Male and female are similar in external morphology except antennae. In male they are, fasciculated with long cilia and very short cilia in female.

Male genitalia (Image 1C). Uncus long, highly sclerotised broad till middle and then narrowing down, apex pointed recurved. Tegumen longer than the uncus, moderately sclerotised with broad arms, inverted v-shaped; valvae symmetrical, weakly sclerotised, setosed, costa strongly produced into a long process, harpe with a pointed process; vinculum longer than tegumen, u-shaped; juxta elongated; Aedeagus (Image 1D) long, relatively thin, apical portion dentate ventrally. Vesica membranous with single, long cornutus.

Female genitala (Image 1E). Corpus bursae oblong, membranous; ductus bursae long, membranous; ostium bursae simple, sclerotized; posterior and anterior apophyses are of equal length, sclerotized; papilla analis oval, heavily sclerotized with setae.

Distribution: India (throughout including Maharashtra), China, Japan, Malaysia, Myanmar, Nepal, Sri Lanka, Taiwan, and Thailand.

Host plants. Ricinus communis, Ficus carica, F. hispida, F. racemosa, F. pumila, F. infectoria, F. religiosa, and Mitragyna diversifolia (ICAR-NBAIR 2020).

DNA barcode studies: In the GenBank a total of 22 sequences of mt COI are available for A. ficus (Table 1), of which nine sequences are from India. Within India, these sequences are from the states of Assam, Maharashtra and Tamil Nadu (all are unpublished data as per GenBank). The current study forms the first published record of DNA barcode for the species A. ficus from India with assigned voucher numbers.

In the preliminary phylogenetic tree generated for the studies, all the mt DNA barcodes formed a monophyletic clade for the species A. ficus (Figure 2) showing genetic distance variance from 0.6% to 1.3%. The clade comprising A. speciosa and A. comorana showed sister relationship with the clade of A. ficus, wherein genetic distance between the species A. ficus and A. comorana was 2.9% and A. ficus and A. speciose was 3.4%. In the present study A. comorana is nested within A. speciosa which suggests either one of the species was wrongly identified ending up in mislabelled sequences or synonymy of these two taxa. Further studies are necessary to resolve the identity and validity of the species A. comorana as the genetic distance between the species A. speciosa and A. comorana is too shallow (0.6–1.7 %).

Evolutionary distances are fundamental in molecular reconstructions including phylogenetic analysis (Nei & Kumar 2000). The nucleotide substitution method is widely used to calculate a reliable genetic difference between pairs of sequences (Nei & Kumar 2000). Since there are limitations with the mt COI gene (Cameron et al. 2004; Hebert & Gregory 2005; Lafontaine & Schmidt 2010), we suggest further studies to comment on the phylogenetic relationships among the species of the genus Asota. Nuclear DNA (n DNA) studies are advocated (Zahiri et al. 2012) to study ancient evolutionary divergence for resolving deeper nodes above species level, having slower mutation rate than mt DNA.

In India, generation of mt COI DNA barcodes for moths is still in a stage of infancy. Recently, Kalawate et al (2020a) have reported the palearctic moth species Olepa schleini Witt et al. 2005 from India with a description of subspecies based on the DNA barcode studies and morphological variations. Additionally, Kalawate et al. (2020b) described three new species along with a subspecies and provided the description of multiple morphotypes of Olepa from India. These studies clearly endorse the utility of DNA barcodes in identification of palearctic species from India (Kalawate et al. 2020a). This technique further avoids  taxonomic inflation by describing morphologically different looking morphotypes as a new species (Kalawate et al. 2020b). Further, DNA barcode studies are expected to alleviate identification of morphologically variant species and uncover the cryptic diversity prevailing within the taxonomic groups. Multigene phylogenetic analysis is warranted to decipher the phylogenetic relationships across the members of the family which are wide spread in distribution range.

 

 

Table 1. Details of the mt COI GenBank accession numbers of Asota utilised in the construction of ML phylogenetic tree.

 

GenBank Accession No.

Locality

Species name as per NCBI

Publication details as per NCBI

1

GU662348.1

Thailand: Chiang Mai

Asota ficus

Unpublished

2

OL630456.1

India: Maharashtra, Nasik, Saptashrungigadh.

Asota ficus

Current study

3

OL630457.1

India: Maharashtra , Jalgaon

Asota ficus

Current study

4

HQ990842.1

Pakistan

Asota ficus

Unpublished

5

KC499430.1

India: Tamil Nadu, Kalkad

Asota ficus

Unpublished

6

MG783922.1

India: Maharashtra

Asota ficus

Unpublished

7

KC499429.1

China: Yunnan

Asota ficus

Unpublished

8

KJ013139.1

India: Assam,

Asota ficus

Unpublished

9

KX860794.1

Pakistan: Punjab

Asota ficus

Ashfaq et al. (2017)

10

MG783907.1

India: Maharashtra

Asota ficus

Unpublished

11

KJ013146.1

India: Nameri NP

Asota ficus

Unpublished

12

JF858113.1

Pakistan

Asota ficus

Unpublished

13

HQ990838.1

Pakistan

Asota ficus

Unpublished

14

JF858114.1

Pakistan

Asota ficus

Unpublished

15

HQ990840.1

Pakistan

Asota ficus

Unpublished

16

HQ990841.1

Pakistan

Asota ficus

Unpublished

17

GU662343.1

Thailand: Chiang Mai

Asota ficus

Unpublished

18

HQ990839.1

Pakistan

Asotaficus

Unpublished

19

HQ990837.1

Pakistan

Asota ficus

Unpublished

20

MG783872.1

India: Maharashtra

Asota ficus

Unpublished

21

MG783923.1

India: Maharashtra

Asota ficus

Unpublished

22

MG783857.1

India: Maharashtra

Asota ficus

Unpublished

23

MG783890.1

India: Maharashtra

Asota ficus

Unpublished

24

MG783877.1

India: Maharashtra

Asota ficus

Unpublished

25

KR070811.1

Kenya: Kajiado North

Asota speciosa

Unpublished

26

KU722731.1

Comoros: Grande Comore

Asota comorana

Unpublished

27

KU722737.1

Comoros: Grande Comore

Asota comorana

Unpublished

28

KR085638.1

Zambia: Victoria Falls

Asota speciosa

Unpublished

29

KR085639.1

Zambia: Lusaka Ridgeway

Asota speciosa

Unpublished

30

GU662438.1

Nigeria: Laeinde

Asota speciosa

Unpublished

31

KJ013107.1

Tanzania: Mbizi forest

Asota speciosa

Unpublished

32

KR736263.1

Nigeria:Oyo

Asota speciosa

Unpublished

33

GU662439.1

Cameroon: North Province

Asota speciosa

Unpublished

34

HM395501.1

Gabon: WoleuNamiTchimble

Asota speciosa

Unpublished

35

HQ573836.1

Gabon: Ogooue-Ivindo

Asota speciosa

Unpublished

36

KR736264.1

Nigeria:Oyo

Asota speciosa

Unpublished

37

KJ013158.1

Ethiopia: Arba Minch

Asota speciosa

Unpublished

38

KJ013170.1

Laos: Nang Phoa

Neochera inops

Unpublished

39

KJ013145.1

Laos: Nang Phoa

Neochera inops

Unpublished

40

KJ013127.1

Laos: Namha protected area,

Neochera inops

Unpublished

41

KC499568.1

Indonesia: Kalimantan Barat

Neochera inops

Unpublished

42

KC499567.1

China: Hainan

Neochera inops

Unpublished

43

KF491909.1

Malaysia

Neochera inops

Unpublished

44

HQ569811.1

Thailand: Nan

Neochera inops

Unpublished

45

HQ569810.1

India: Meghalaya

Neochera inops

Unpublished

46

HQ569809.1

VietNam: Tam Dao

Neochera inops

Unpublished

47

GU662423.1

Thailand: Chiang Mai

Neochera inops

Unpublished

48

GU662331.1

Thailand: Chiang Mai

Neochera inops

Unpublished

49

JN401278.1

Japan

Neochera inops

Zahiri et al. (2012)

50

HQ569812.1

Malaysia: Sarawak

Neochera inops

Unpublished

 

 

For figures & image - - click here

 

 

References

 

Anonymous (2016). Alert issued for fever caused by tiger moth. http://timesofindia.indiatimes.com/articleshow/51869389.cms?utm_source=contentofinterest&utm_medium=text&utm_campaign=cppst assessed 01.12.2020.

Ashfaq, M., S. Akhtar, M.A. Rafi, S. Mansoor & P.D. Hebert (2017). Mapping global biodiversity connections with DNA barcodes: Lepidoptera of Pakistan. PLoS ONE 12(3): e0174749. https://doi.org/10.1371/journal.pone.0174749

Bayarsaikhan, U., N. Sol-Moon & B. Yang-Seop (2016). Review of the subfamily Aganainae (Lepidoptera, Erebidae) from Cambodia. Journal of Asia-Pacific Biodiversity 9(2): 219–229. https://doi.org/10.1016/j.japb.2016.02.010

Cameron, S.L., K.B. Miller, C.A. D’Haese, M.F. Whiting & S.C. Barker (2004). Mitochondrial genome data alone are not enough to unambiguously resolve the relationships of Entognatha, Insecta and Crustacea sensu lato (Arthropoda). Cladistics 20(6): 534–557. https://doi.org/10.1111/j.1096-0031.2004.00040.x

Common, I.F.B. (1990). Moths of Australia. E.J. Brill and Melbourne University Press, New York.128 pp.

Daniel, F. (1943). Beiträge zur Kenntnis der Arctiidae Ostasiens unter besonderer Berücksichtigung der Ausbeuten H. Höne’s aus diesem Gebiet (Lep. Het.). II Teil. Hypsinae, Micrarctiinae, Spilosominae, Arctiinae. Mitteilungen der München Entomologischen Gesellschft 33: 673–759.

Fibiger, M. & J.D. Lafontaine (2005). A review of the higher classification of the Noctuoidea (Lepidoptera) with special reference to the Holarctic fauna. Esperiana 11: 7–92.

Folmer, O., W.R. Hoeh, M.B. Black & R.C. Vrijenhoek (1994). Conserved primers for PCR amplification of mitochondrial DNA from different invertebrate phyla. Molecular Marine Biology and Biotechnology 3(5): 294–299.

Hampson, G.F. (1892). The fauna of British India including Ceylon and Burma, Moths - Volume 1. Taylor and Francis, London, 504 pp.

Hebert, P.D., S. Ratnasingham & J.R. deWaard (2003a). Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings Biological sciences 270 (Suppl. 1): S96–S99. https://doi.org/10.1098/rsbl.2003.0025

Hebert, P.D., A. Cywinska, S.L. Ball & J.R. deWaard (2003b). Biological identifications through DNA barcodes. Proceedings Biological Sciences 270(1512): 313–21. https://doi.org/10.1098/rspb.2002.2218

Hebert, P.D.N. & T.R. Gregory (2005). The promise of DNA barcoding for taxonomy. Systematic Biology 54 (5): 852–859. https://doi.org/10.1080/10635150500354886

Holloway, J.D. (1988). The Moths of Borneo, part. 6: family Arctiidae, subfamilies Syntominae, Euchromiinae, Arctiinae; Noctuidae misplaced in Arctiidae (Camptoloma, Aganaidae). SouthdeneSdnBhd, Kuala Lumpur.

ICAR-NBAIR (2020). Asota ficus. https://www.nbair.res.in/Databases/Databases/insectpests/Asota-caricae php+&cd=12&hl=en&ct=clnk&gl=in   accessed 08.xii.2020.

Inoue, H., S. Sugi, H. Kuroko, A. Kawabe & M. Owada (1982). Moths of Japan. Kodansha, Tokyo, 344–405 pp.

Kalawate, A.S., S. Pawara, A. Shabnam & K.P. Dinesh (2020a). DNA barcode reveals the occurrence of Palearctic Olepas chleini Witt et al., 2005 (Lepidoptera: Erebidae: Arctiinae) from peninsular India with morphological variations and a new subspecies. Journal of Threatened Taxa 12(9): 16143–16152. https://doi.org/10.11609/jot.5596.12.9.16143-16152

Kalawate, A.S., K.P. Dinesh & A. Shabnam (2020b). DNA barcoding unravels three new species and a subspecies of Olepa Watson, 1980 (Lepidoptera, Erebidae, Arctiinae) from India, with morphotypes. Journal of Insect Biodiversity 19(2): 44–60.  https://doi.org/10.12976/jib/2020.19.2.2

Kitching, I.J. & J. Rawlins (1998). The Noctuoidea, pp. 355–401. Kristensen, N.P. (ed.). Handbook of Zoology, Lepidoptera, Moths and Butterflies, Vol. 1. Evolution, Systematics, and Biogeography. W. de Gruyter, Berlin. 

Lafontaine, J.D. & B.C. Schmidt (2010). Annotated check list of the Noctuoidea (Insecta, Lepidoptera) of North America north of Mexico. ZooKeys 40: 1–239. https://doi.org/10.3897/zookeys.40.414

Nei, M. & S. Kumar (2000). Molecular Evolution and Phylogenetics. Oxford University Press, 333 pp.

Robinson, G.S. (1976). The preparation of slides of Lepidoptera genitalia with special reference to the Microlepidoptera. Entomologist’s Gazette 27(2): 127–132.

Scoble, M.J. (1992). The Lepidoptera. Form, Function and Diversity. Oxford University Press, Oxford, 404 pp.

Seitz, A. (1914). The Macrolepidoptera of the world. II. Division: Fauna Exotica, A. Kernen, Stuttgart, 10: 105–290 (Bombyces and Sphinges of the Indo-Australian Region). https://doi.org/10.5962/bhl.title.9400

Silvestro, D. & I. Michalak (2012). raxmlGUI: a graphical front-end for RAxML. Organisms Diversity & Evolution 12(4): 335–337.

Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei & S. Kumar (2011). MEGA5: Molecular Evolutonary Genetics Analysis using Maximum Likelihood, Evolutionary Distance and Maximum Parsimony Methods. Molecular Biology and Evolution 28(10): 2731−2739. https://doi.org/10.1093/molbev/msr121

Wills, P.J., M. Anjana, M. Nitin, R. Varun, P. Sachidanandan, T.M. Jacob, L. Madhavan, R.V. Thampah & K.K. Varma (2016). Population explosions of Tiger Moth lead to Lepidopterism mimicking infectious fever outbreaks. PLoS ONE 11(4): e0152787. https://doi.org/10.1371/journal.pone.0152787

Zahiri, R., I.J. Kitching, J.D. Lafontaine, M. Mutanen, L. Kaila, J.D. Holloway & N. Wahlberg (2011). A new molecular phylogeny offers hope for a stable family level classification of the Noctuoidea (Lepidoptera). Zoologica Scripta 40(2): 158–173.

Zahiri, R., J.D. Holloway, I.J. Kitching, J.D. Lafontaine, M. Mutanen, & N. Wahlberg (2012). Molecular phylogenetics of Erebidae (Lepidoptera, Noctuoidea). Systematic Entomology 37(1): 102–124. https://doi.org/10.1111/j.1365-3113.2011.00607.x