Papilio+demoleus+malayanus

The Lime Butterfly Papilio demoleus

.  Wanted for crop damage and invasion of other habitats. ( Beware! my children are hungry!) 2. Introduction (caterpillar pictures)

The Lime Butterfly, // Papilio demoleus // is a commonly widespread swallowtail butterfly found in open areas that spans from South Asia to Australia(Lewis,2012; Smith& Vane-Wright,2008) The subspecies // Papilio demoleus malayanus // is found mainly in the region of Southeast Asia and can usually be sighted visiting flowers such as // Bougainville spp //. and // Ixora spp //. that are often found in the gardens of Singapore residential.

The name Lime Butterfly is often derived from its hostplant that its caterpillars/larva feeds on //Citrus spp.// plants such as Key Lime (//Citrus// //aurantifolia//). Due to the import of kumquat, another //Citrus spp.// plant during Chinese New Year season in Singapore, you can see them more often as there are more food for their caterpillars to feed on.

Although the adult lime butterfly may look pretty in our eyes, the caterpillars have a large appetite is considered as minor garden pest in Singapore. For countries with large lime plantation industry, these caterpillars can pose a serious threat in the Old World such as India and Saudi Arabia. They are aggressive pests and spread across foreign countries very quickly. Recent findings have shown that this species has spread to the New World such as Dominican Republic and Puerto Rico, threatening the livelihood of farmers that depends on lime cultivation. Therefore the lime butterflies pose as a serious threat to these countries.

Gurrero et al.,2004)

In this species page, we will try to understand more about the biology of //Papilio demoleus// in order to show the impact lime butterflies have on humans as an invasive species and an economic pest. Taxonomic information will be provided the help in identification of the butterfly through its __description__, __diagnosis__ and __distribution__ throughout the world. Lastly, __systematic information__ about the species and its related outgroups will be provided show the comparison between //Papilio demoleus// and other similar looking species, of which, some are rare and can only be found in Madagascar.

=Life Cycle=

Overview
As all butterflies go through holometabolous development or complete metamorphosis, that is there are 4 stages in the life cycle of the lime butterfly. However it is difficult to distinguish lime butterfly from other species when it is larvae or pupa stage. In Singapore, the larval and pupa stages of lime butterfly often resembles that of Common Mormon (//__Papilio polytes__//) and Great Mormon (//__Papilio memnon__//). Therefore upon spotting of these caterpillar, we can guess they belong to these 3 species. The following description of the life cycle is based on the book “Caterpillars of Singapore’s Butterflies” by Horace Tan and Khew Sin Khoon.

Egg stage
Eggs are laid singly on young leaves or on the underside of the leaves as well as the surface of twings on lime host plant( Atluri et al 2002). The eggs laid are spherical and smooth edged. Due to the tropical climate of Singapore, the time for embryo development is very short, about 2-3 days.

Larval stage

 * //<Picture of 3rd instar caterpillar)//**

During its early stages, the caterpillar is **light brown in colour with white markings**, closely resembling a bird’s droppings as camouflage. As it grows, it will moult or shed its exoskeleton. Every time it moults, it will grow from the 1st instar to the 2nd instar (see instar for definition). From the 1st to the 4th instar, it appearance does not change much other than its size. The white markings in the following 3rd instar specimen (meaning it has shed its skin 2 times) is V-shaped that covered the top and the side of its body. A row of spines can also observed on the top side of the body. As the caterpillar grows, the size of the white marking will extend and the colour will shift from light brown to dark brown or black. Refer to butterflycircle.com for more details

On the 5th instar, the caterpillar changes to yellowish green / green in colour with no spines, closely resembling the colour of its __lime host plant__. In the middle or 3rd thoracic segment, there are 2 lateral false eyespots with a yellowish brown band spanning across the segment. Base on this specimen, the pillar has brown marking on both sides of its body. It is during this latter stage that the larva can die very easily due to parsitoid wasps

Pupal stage
//The pupae take the colour of either green or brown// depending on the surface that it pupate on. Pupating on non-woody or smooth surface results in a green pupae (cite). Pupating on woody or brown surface gives a brown pupae(cite). The pupa is supported by the silk girdle across the __metathorax__ and and attachment on __cremaster__. __(see anatomy)__

Description Please refer to the description section for more information about the butterfly
Behaviour

= 3. Classification =

Name

Species name //Papilio demoleus// Linnaeus 1758
 * Binomial:
 * // Papilio demoleus malayanus // Wallace, 1865

Synonyms - Papilio demoleus Linnaeus,1758
 * Princeps demoleus Linnaeus 1758
 * Papilio erithonius Cramer,1779
 * Papilio epius Fabricius,1793
 * Papilio demoleinus Oberthur,1879

- Papilio demoleus malayanus Wallace,1865
 * Papilio demoleus annamiticus Fruhstorfer,1901

Vernacular
 * Common Lime Butterfly
 * Lemon Butterfly
 * Lime Swallowtail
 * Chequered Swallow tail
 * Citrus Swallowtail || media type="custom" key="24408240" ||

3.1 Taxonavigation
Animalia Arthropoda Insecta Lepidoptera Ditrysia Papilionoidea Papilionidae Papilioninae Papilionini Papilio demoleus malayanus

Papilio demoleus group (Lime Swallowtails) Papilio demoleus group comprises the following 5 species that are considered lime swallowtails and has the same wing pattern of each other (Zakharov,2004a; Smith & Vane-Wright,2008). See for visual photo

1) Papilio demoleus Linnaeus,1758 2) Papilio morondavana Grose Smith, 1891 3) Papilio grosesmithi Rothschild,1926 4) Papilio demodocus Esper,1799 5) Papilio erithonioides Grose Smith, 1891a

Papilio demoleus subspecies The number of subspecies has decrease from 2004 to 2008. Based on the most recent paper by Smith& Vane-Wright (2008), there are five subspecies instead of six as P.d.libanius Frustorfer is synonymous to P.d. demoleus,Linnaeus.

1) P.d .malayanus Wallace, 1865 2) P.d .demoleus Linnaeus, 1758 3) P. d. sthenelus, Macleay, 1826 4) P.d. stenelinus Rothschild, 1895 5) P.d. novoguineensis Rothschild,1908 6) P.d .libanius Fruhstorfer, 1908 (= P.d.demoleus)

Differentiation of Papilio demoleus malayanus from other members of the Papilio demoleus group species and Papilio demoleus subspecies can be found in the Diagnosis section. The systematic issues subspecies will be discussed in the Taxonomy section. = 4. General Anatomy =

4.1 Adult wing anatomy
Identification of butterfly species is heavily based on morphology and in particular the wing anatomy. The venation pattern is based on Comstock-Needham system where the venation categories are based on insect instead of Lepidotpera i.e butterflies and moths (Miller,1969). Venation of the wings, starting from the anterior are as follows (Miller, 1969) :-
 * Subcosta (Sc),
 * Radius (R1 - R5),
 * Medius (M1- M3),
 * Cubitus (Cu),
 * Anal veins (1A,2A)

4.2 Adult body anatomy
= 5 General Description =

So what does the lime butterfly Papilio demoleus look like?

Butterflies are generally classified based on their wing patterns which are distinct between species. The description pertains to the species Papilio demoleus. Description of the subspecies can be found in the diagnosis section. Butterflycircle.com also provides concise information about Papilio demoleus malayanus in Singapore.

5.1 Dorsal (Upper side) Wing


1) Dorsal side of fore wings and hind wings are brownish-black in color with a chain of irregar light yellow spots along the discal, submarginal and marginal regions

2) Hind wings has a black costal eyespot near apex that gradually spreads over the blue scales shaped as a crescent. (Bascombe, 1999; Braby, 2000; Corbet, 1978)

3) In space 1b on the dorsal hind wing, there is a red tornal spot for both sexes. Different genders usually have distinctly dissimilar color composition (Bascombe, 1999; Braby, 2000; Corbet, 1978). See Differentiation of gender

5.2 Ventral(Underside) Wing
4) Forewings are brown-black in color with similar spot arrangement to the upper side.

5) Forewings are yellow/dull yellow in color with more extensive light yellow spots compared to the dorsal side

5) A series of dull-yellow spots are located on the sub apical (below the apex) of the fore wing (Bascombe, 1999; Braby, 2000; Corbet, 1978) 6) Underside of hindwings has a series of elongated central (post-discal) orange spots edged with iridescent blue and black. Costal spot near apex is orange edged with blue and black Edge of the hindwings are wavy. (Bascombe, 1999; Braby, 2000; Corbet, 1978)

5.3 Sexual dimorphism
1) Tornal spot pattern Males – tornal spot on upper side hind wing is mostly orange with little blue scale  Females - tornal spot on upper side hind wing is partially orange with more blue scales (Braby, 2000)

2) Ventral hind wing color Males – underside of hind wing are light yellow in the discal and submarginal region  Females – underside of hind wing is dull yellow in the discal and submarginal region  In many specimens the light yellow spots are darken to dull yellow with age(Bascombe, 1999).  Of the 8 females out of 25 eggs, 2 have light yellow markings and 6 have dark yellow markings on the ventral hindwings (Bascombe, 1999).  However, gender differentiation are most reliably determined by the presence of claspers (Bascombe, 1999).



3) Shape of terminal abdomen segment Male – sharper abdomen shape with large end  Female – round abdomen shape.

= 6. Diagnosis =

This section provides a brief summary of indicators that differentiates papilio demoleus from other species of lime butterfly. There are five species in the demoleus group.

6.1 Differentiation of P. demoleus from other lime swallowtails
Based on the dichotomous keys provided by Smith & Vane-Wright(2008).There are 2 essential character states that differentiate Papilio demoleus from other lime swallowtails



P.demoleus vs P.morondavana & P.grosesmithi P. demoleus – tail on hindwing vein M3 is rudimentary or very short, less than twice as long as those of neighbouring veins P.morondavana, P.grosesmithi – hind wing vein M3 extended to form distinct tail at least 3 times as long as extensions of neighbouring veins P. demoleus vs P. erithonioides & P.demodocus P.demoleus - forewing upperside cell M1 with post-discal mark absent P.erithonioides & P.demodocus – forewing upperside cell M1 with post-discal mark present

6.2 Differentiation of P.d.malayanus from other subspecies
Differentiation of P.d.malaynus from the other subspecies based on the dichotomous keys



1) Pre-apical yellow spot on fore wing discal cell

P.d.malayanus & P.d.demoleus – Forewing discal cell with 2 separate pre-apical yellow spots, very rarely less than 0.5mm apart. (Asia and Wallacea)

P.d.sthenelus & P.d.sthenelinus & P.d.novoguineensis - Forewing discal cell with pre-apical spots confluent ; if separated, very rarely more than 0.5mm apart

2) Length of yellow postdiscal marks on cell 1A

P.d.malayanus - Yellow post discal marks in forewing cell 1A rectangular, rarely less than 6mm in width

P.d.demoleus - Yellow postdiscal mark in forewing cell 1A subquadrate usually about 4mm or less across. Rarely more than 5mm



3) Length of yellow band the hindwing discal cell

P.d.malayanus - Pale band of hindwing discal cell of male relatively ‘broad’, generally extending to root of vein CuA1

P.d.demoleus - Pale band of hindwing discal cell of male relatively ‘narrow’, generally extending to half way between the roots of veins CuA1 and CuA2 = 7. Distribution =

7.1 Distribution in the World
Papilio demoleus malayanus appear to widely occur in Southeast Asia region and a continue spread southward across the Indonesia Archipelago and the Papua Province. fujioka (1997). Recent introduction into the New World may also been observed as specimen collected from Jamaica (NAPPO,2006) Dominican Republic (Guerrero et al.,2004) Puerto Rico (Homziak & Homziak, 2006) is observed. Burma, Thailand, Cambodia, Laos, central and southern Vietnam, Sabah and the Malay Peninsula. media type="custom" key="24408172"

7.2 Distribution in Singapore
Generally, Papilio demoleus malayanus can be found in most open spaces of urban Singapore. The planting of nectar flowers around residential estate help to attract these butterflies to the areas. In old estates such as Hougang areas, many residents would plant lime such as Key lime (Citrus aurantifolia) at their backyard, providing favourable host plants for the larvae of lime butterflies to feed on. The following map shows some of my personal recording in collecting lime butterfly specimens.

The time for the emergence of the adult usually occurred at about 0800H (Corbet and Pendle bury, 1992) and active until 1200H. Occasional sighting of the adults can also be found in the late afternoon as well. media type="custom" key="24407888" = 8. Taxonomy and Systematics =

8.1 Etymology
“Papilio” is derived from the Latin term “papilionem” and the French term “papillon”. Both term mean “butterfly, moth”. There is no clear meaning behind the species demoleus coined by Linnaeus as the name ”demoleus” refers to a greek man who had been slain by Aeneas, a Trojan involved in the war of Troy. “Malayanus”,as the name suggested refers to the Malay peninsula where Wallace establish the subspecies as a local form in Southeast Asia (Smith& Vane-Wright,2008)

8.2 Types series


The Male lectotype of Papilio demoleus malayanus is found in the British Museum of Natural History in London, United Kingdom. The specimen acquired was from habitat in Singapore. The Lectotype is a specimen from Wallace’s private collection (Smith & Vane-Wright ,2008)

8.3 Phylogeny relationships


Fig. 8.2. One of two equally parsimonious trees (TL 52533, CI50.548, RI50.554) based on combined dataset. Most parsimonious trees were different only in some outgroup rearrangements. Bayesian analysis produced tree with almost identical topology (-lnL5 17024.14) with different position of Papilio anactus shown by a circled star. Histograms above nodes show partitioned Bremer supports,numbers under nodes indicate bootstrap proportions, total Bremer support, and clade posterior probabilities from Bayesian analyse (Diagram from Zakharov et al.,(2004a) Permission not acquired but within the limits of Fair Use)

Cladistic analysis done by Zakharov et al. (2004ainvolves the molecular phylogeny and morphological characteristic to test for phylogenic relationship between the species in the P.demoleus group and outgroups of the Papilio genus. Based on the cladogram derived from the combined data (COI, COII, Wg, EF1a genes and morphology), here is a strong support between the species in the P.demoleus group, showing that P.demoleus group is monophyletic.

Shift of basal position from P.demoleus or P.demodocus to P.morondavana. The strongest evidence indicates that a clear sister relationship between P.demodocus and P.erithonioides is establish with P.demoleus likely to be their sister taxon. P.grosesmithi is the sister taxon of the 3 species and P.morondavana takes the basal position. The data no longer support that P.demodocus or P.demoleus is the most basal species. The finding urges that more emphasis should be placed on P.morondavana in its conservation effort as it appears to hold the oldest lineages in P.demoleus group.

Does Papilio demoleus group belong to the subgenus Princeps? Based on Hancock (1983), the P. demoleus group belonged to the subgenus Princeps. Yet a more recent evidence on molecular phylogeny of Papilio (Zakharov et al.,2004a) indicates that Princeps is paraphyletic and the demoleus group shown currently occupy between the 2 subdivision of Papilio genus, Menelaides and Achillides, where Menelaides is the sister taxon to P.demoleus group. Therefore P.demoleus has been shifted out from the subgenus Princeps. As the cladogram suggested that P.(Menelaides) helenus is one closet outgroup to P.demoleus group, it I further supports the claim that P.demoleus group is not part of Princeps.

8.4 Biogeography
P.demoleus and its related species may originate from Madagascar. The approximate age of divergence of P.demoleus group is estimated to be about 16.8 million years. According to estimated substitution rate for COI and COII genes(Zakharov et al, 2004b), it is likely that the ancestor of P.mordavana and the rest of the P.demoleus group diverged between 14.1 and 10.6 million years ago. Another split of the ancestor for P.grosesmithi and its sister group occurred between 13.5 and 10.3 million years ago(Zakharov et al.,2004b). Dispersal-vicariance analysis suggests that the origin and speciation of P.demoleus group occurred within Madagascar. Dispersal of P.demoleus into Australia and Asia was dated to around 10.1- 7.8 million years ago while dispersal of P.demodocus into Africa was 7.3-5.6 million years ago (Zakharov et al.,2004b). Fig. 8.3 Sampled localities and distribution of 27 unique haplotypes identified from 50 ingroup individuals of Papilio group from 825-bp fragment of the COI gene. Arrows indicate prevailing wind and currents in the Indian Ocean that are influenced by the Asiatic monsoon and trade winds (Walter 1970), including North Equatorial Current (I), Mozambique Current (II), Alguhas Stream (III), West Wind Drift Current (IV), West Australian Current (V), and South Equatorial Current (VI). (Diagram from Zakharov et al.,(2004a). Permission not acquired but within the limiits of Fair use)

Mechanism of the dispersion It is suggested that long-dispersal events across the Indian Oceans occurred several times. The mechanism of such transmarine dispersal was made possible from the influence of Asiatic monsoon season and trade winds. The westward dispersal of the lime swallowtails most likely occurred on Northern Moxambique Channel and has occurred on other lepidopteran species on several occasions (Pierre, 1992; Torres et al, 2001). The prevailing easterly trade winds and ocean current is likely to be responsible for the dispersal of lime swallowtails from Asia to western Indian Ocean. Dispersal from southern Madagascar and southern Africa is likely to be facilitated by West Wind Drifts.

Rebuttal of dispersal-vicariance analysis It is also possible that the dispersal-vicariance analysis is incorrect in that the distribution of lime swallowtails is not parsimonious for such feat. It is possible that the extinction of intermediates would complicate the reconstruction of dispersal and deny the Malagasy origin of the lime swallowtails

Dispersal of P.demoleus For P.demoleus, it is likely that its origin in Saudi Arabia and Iran comes from the introduction of Citrus in the 10th century (Wiltshire ,1945; Larsen,1977) instead of Southeast Asia and Eastern Asia. The introduction may have come from closer sources such India subcontinent as there seem to be a moderate divergence (1%) between the samples from Iran and samples from Southeast Asia and East Asia.

P.d.sthenelus as separate species from other P.demoleus subspecies There is almost a significant divergence between Oriental subspecies (P.d.malayanus & P.d.demoleus) from Australian subspecies (P.d.sthenelus) in COI gene (Tsao & Yeh,2008). Given that there is a shift in larval host plant from Rutaceae to Fabaceae; P.d.sthnelus might be grouped as a separate species. Its isolation of ancestral population of P.demoleus in Oriental and Australian region might have been very long, ranging from 3.24 to 4.24 million years. Thus, P.d.sthenelus may represent the oldest lineage in P.demoleus. However, more detailed are required to confirm the following observation.

= 9.Human Impacts =

9.1 P.demoleus as a global pest.
With a large dispersal range, high reproductive range and wide range of larval host plants, the spreading of P.demoleus population are often fast and rampant. Based from Atluri et al. (2002), the P.demoleus is capable of reproducing up to six generation annually in India and has a preference for 19 species of citrus varieties including cultivated species (Guerrero et al.,2004). There is also a positive correlation of P.demoleus abundance with seasonal increase in temperature and relative humidity in West Bengal, India. Originally located in the Old World, a recent record of P.demoleus species which is likely to be the subspecies from Southeast Asia or P.d.malayanus was found in Dominican Republic in 2004(Guerrero et al.,2004; Eastwood et al., 2006), followed by Puerto Rico in 2006 (Homziak & Homziak, 2006)

Beginning in the 1970s, invasion of P.demoleus have already occurred in the islands of Java, Borneo, Philippines and Sumatra. The invasion was facilitated by deforestation and the increase availability of citrus grove (Matsumoto, 1996, 2002). P.demoleus is also known to plague citrus cultivation in Saudi Arabia and Iran (Badawi, 1981). Narayayamma et al. (2001) has also reported that up to 83% defoliation of young grove tress in Andra Pradesh, making P.demoleus an economic pest in India as well. The recent invasion of P.demoleus happened to have occurred by a single introduction in 2004 in Dominican republic as evidence showed that there is a lack of genetic variation among the specimens sampled in Dominican Republic (Eastwood et al.,2006). It is however no known how the P.demoleus reached the Dominican Republic. Homziak & Homziak (2006) suggested that the invasion of P.demoleus in Puerto Rico in March 2006 is likely originated from the Dominican Republic.

With the presence of favorable climatic conditions, favorable host plant and potential lack of predators, the establishment of P.demoleus in Dominican Republic is inevitable (Guerrero et al.,2004)

9.2 Economic Importance
The invasion of P.demoleus into the New World such as Dominican Republic and Puerto Rico has raised alarm with regards to the potential threat this species may have on the agricultural industry of United States and Hispaniola. Based on estimates from the Caribbean cooperative citrus association (CARICOM), the production of Citrus products amounts to an approximate 1,100,000 in CARICOM countries such as Cuba, Dominican Republic and Haiti, generating an amount of US$61 million annually (Homziak & Homziak,2006) Furthermore, small citrus farms make up between 93 and 98 percent of all CARICOM farms. (Homziak & Homziak, 2006) Therefore the industry is critical to the economic survival of many small farmers (Homziak & Homziak, 2006). Based on National Agricultural Statistic Service (2004), the value of Florida citrus crop exceed, US 1.5 billion dollars and United States total worth was more than US$2.6 billion dollar. With the population of P.demoleus established in Dominican Republic, it is likely that the population can easily spread through the West Indies and surrounding areas, including the United States where impact on the agricultural industry could be catastrophic. (Guerrero et al.,2004)

1) Biological control
Many literatures have proposed biological control of P.demoleus through the use of predators and natural pesticide. Biological predators that could be utilized to control the population - Larva parasitoids - Pupal parasitoids - Adult predators
 * - Egg parasitoids
 * o Ooencyrtus malayensis (Hymenoptera: Encyrtidae), Thailand 3
 * o Tetrastichussp.(Hymenoptera: Eulophidae), Thailand 3
 * o Encyrtid (Hymenoptera), Jamaica
 * o Chalcidoid (Hymenoptera), Jamaica
 * o Erycia nymphalidophaga (Diptera: Tachinidae), Thailand 3
 * o Apanteles papilionis, (Hymenoptera: Braconidae)India 3
 * o Apanteles sp., (Hymenoptera: Braconidae),India 3
 * o Bracon hebetor (Hymenoptera: Braconidae), India. 3
 * o Brachymeria sp. (Hymenoptera: Chalcididae)
 * o Pteromalus puparum (Hymenoptera: Pteromalidae).
 * o Cantheconidea furcellata (Hemiptera: Pentatomidae)
 * o Reduviidae sp. (Hemiptera)
 * o Spiders
 * o Sphecids (Hymenoptera)
 * o Chameleons and birds || [[image:pdm_braconid_parasitoid.jpg caption="Bracoid parasitoid feeding off P.demoleus larvae (Photo by J.M. Garg,(2009)from Wikicommons)"]] ||

Biopesticides Bacillus thuringiensis and Beauveria bassiana as well as neem seed kernel extract and azadjrachtin were shown to have effects on P.demoleus in India. (Lewis,2009)

Neem based insecticides( multineem and neemark) could serve to alter the life-processing behavior that the insect can no longer feed, breed or undergo metamorphosis. The insecticide could induce defecation and sporadic jerking of the body. Heavy pigmentation in the 5th instar larvae, disappearance of red spot and change in wing shape, significant reduction of oocytes number are some of the effects on P.demoleus (Pandey et al., 2011)

2) Non-biological control
Regulation of fatty acid has known to influence the growth development of insects (Methat, 1970). Methat (1970) has demonstrated that the use of fatty acids could serve to inhibit the growth of caterpillar larva. Treatment of lime leaves with oleic, linoleic and erucic acid at concentration higher than 0.25% proved to be toxic to the larva and inhibit pupation. (Methat, 1970)

= 10. References =

Alturi, J. B., Ramna, S. P. V., Reddi., C. S.