CURRENT STATUS AND CHALLENGES FOR CONSERVATION AND SUSTAINABLE USE OF BIODIVERSITY

286 Current status and challenges for conservation and sustainable use of biodiversity butterflies” exhibit aposematically colored wing and brown, black and white from the upperside patterns that are often associated either with hindwing area. Further, underside (ventral) wing Batesian or Müllerian mimicry species to form a scales are also studied. “mimicry ring”(Nishida, 2017). Analysing the Danaus chrysippus butterfly colour patterns does not A total of 159 varieties of wing scales were studied. only enrich our understanding of inter/intra-specific Among them upperside (dorsal) forewing area communication and the evolution of exaggerated comprises of 27 scales from brown area followed by signaling of butterflies, it can also be the source of 22 scales from black and 12 scales from white. new photonic devices. However, upperside hindwing area comprises of 18 scales from brown, 24 scales from black and 27 Materials and Methods scales from white. 29 wing scales from the underside (ventral) area of the wings was also been examined The butterflies of Danaus chrysippus was collected (Table 1, Fig. 1).Focusing on the brown colored area from the premises of Sree Narayana College, Kollam of the upperside forewing enumerate several dark to (8˚525̍ 5̎N76˚36̍4̎ E) by using handheld insect net. pale brown coloured scales (Plate B1: 1-24, Plate Orange, black, brown and white colours were B2: 25-27). The anatomy of the wing scales appears observed from the wings. The scales are removed to be basically identical with an upper lamina from the wing surface as per the standard method of (abwing) and a lower lamina (adwing) together with Grodnitsky and Kozlov (1991). Scales were a stalk. This category has more or less lower lamina extracted from each region of wing separately that (adwing). The upper lamina (abwing) is of structured shows varied colors into a glass slide. A drop of as well as unstructured and is mainly broader xylene was used for fixation of scales and the compared to the adwing. All scales except scale samples were studied under the light microscope. no:24 (Plate B1) and scale no:25,26 (Plate B2) are The measurements of the scales were calculated structured. The abwing of the structured scales using micrometry. Microscopic photography was possess several buds or tooth. The number of used for taking the photomicrographs of the dentations varies for each scale. Some scales prepared scale sample for result analysis. contains two buds or tooth (Plate B1: 4, 23) while some have three buds (Plate B1: 1,5,10,17,18,20,27). Results Others have more than three buds(Plate B1: 2,3, 6, 7, 8, 9, 11, 12, 13,14, 15, 19, 21, 22). However, some Wings are characterized by having a brighter orange scales lack dentations(Plate B1: 24; Plate B2: 25,26). color on the upperside richer than the underside. The Some scales shows pale brown colouration which is apical half of the forewing is black with a white expressed in (Plate B1: 1, 2, 3, 4, 5, 7, 9, 11, 14, 15, band. The hindwing has three black spots in the 17, 19, 20, 21, 22, 23, 24; Plate B2: 25,26,27) while center and are bordered in black and outlined with the other scales possess dark brown colour (Plate semi- circular white spots. Underside is dull and B1: 6,8,10,12,13,16,18). The dimensions of each possesses the similar pattern of upperside. Wingspan scale vary in their length and width. Some scales ranges from 2 -4 cm for forewing and 2-3 cm for visually appears to be similar but there dimensions hindwing (Plate B).On investigating the upper and vary slightly (Plate B1: 8, 13). These slight changes underside wing scales of the ‘Danaus chrysippus’ are important for the normal functioning of the revealed the presence of numerous structured and butterfly wing. Scales can be of same length but they unstructured scales. The scales are isolated from the vary in their thickness (Plate B1: 4, 10). Scale no: 9 differently coloured wing areas namely brown, black and 12 (Plate B1) appears to be similar but there and white from the upperside (dorsal) forewing area Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

A preview on the wing scales of danaus chrysippus using light microscopy 287 color and dimensions often vary. The scale dentations(Plate B3: 66; Plate B4: dimension of this area ranges from 78.8µ-110.32µ in 68,69,70,73,74,77,78). Odd scales are obtained length and 63.04µ-90.62µ in width. Moving to the which is deeply paler than the ground color(Plate second region, that is the black colored region of B4: 71,76,77). The hindwing area brown scales are forewing area of upperside (dorsal) wing produces less in number that makes the hindwing appears dull several black colored scales together with some odd with comparison to the forewing. The dimension white scales (Plate B2: 28-45; Plate B3: 46-49). ranges from 86.68µ-98.5µ in length and 66.98µ- Here the abwing is structured with two or more 74.86µ in thickness. Here all the scales follow the dentations. The base of the adwing possesses a stalk basic anatomy of having an adwing, abwing and a and the intensity of black color for each scale differ. stalk. The black colored region on the upperside Some scales are deeply dark colored (Plate B2: (dorsal) hindwing area shows 24 variety of scales as 28,29,32,33,37,39,40,44; Plate B3: 47) while some that of the forewing scales is only 22. Here the are paler (Plate B2: 31,35,41,42,45; Plate B3: 46). scales appear paler and dull unlike the forewing Isolate scales observed are a few white scales (Plate black scales. In this area, the majority of the scales B2: 30, 36) are also striking. have their abwing devoid of dentations (Plate B4:80,82,83,86,87,88,89,90,91; Plate B5: 93,95,96, The dimensions of scale ranges from 78.8µ-110.32µ 97,99,100,101,102,103). A very few scales have in length and 63.04µ- 78.8µ in width. Here the structured abwing (Plate B4: 81, 84, 85; Plate B5: majority of the scales are broader(Plate B2: 28,30, 92, 94, 98). In these scales, it is clear that the abwing 31, 32, 33,34,36,37,38,39,40,43,44,45; Plate B3: of some scales is more broader while the adwing 48,49) while a few are longer (Plate B2: 29,41,42). seems to be narrow (Plate B4: 81,82,83,89;Plate B5: The scale count is less when compared to the scales 95,100). This region also has several white scales of brown colored region on the dorsal forewing. oddly (Plate B5: 98, 99). Here, the black and white Thirdly, the scales of the white spotted region of scales are intermixed in this region. Hindwing black upperside (dorsal) forewing are examined. It shows a colored area contains several scales that are not very few structured scales (Plate B3: 50-61) except observed on the forewing area and vice versa. The scale no:57 (Plate B3) which is a single round dimensions of the scales range between 86.68µ- unstructured scale. Here the abwing possess several 98.5µ in length and 63.04µ-74.86µ in width. dendations and the adwing seems to be narrow compared to the abwing. On close examination it is Examining the white portion of the upperside observed that majority of the white scales are short (dorsal) hindwing shows several white colored and broader (Plate B3: 52,53,54,55,56,58,59,61) scales that are very striking. White scales also while a few are long and narrow (Plate B3: contain structural as well as unstructured abwing. 50,51,60). The white scales are very attractive to the Some are glassy type and are transparent (Plate B5: eye and its dimensions ranges from 78.8µ-102.44µ 112,115; Plate B6: 116). Scale no: 105 (Plate B5) is in length and 59.1µ-78.8µ in thickness. Investigation an odd scale observed in this area. However, the on the upperside(dorsal) hindwing area focus on number of white scales observed in the hindwing several brown, black and white colored scales. area is more in number compared to the forewing. Brown colour region contains a total of 18 scales The white scales of hindwing area possess more that differ from each other. Here the scales are paler transparency than the upperside (dorsal) forewing than the forewing area scales. Some scale possess (Plate B5 :112, Plate B3: 51). Basically, all scales dentations (Plate B3: 62,63,64,67; Plate B4: have narrow adwing compared to adwing but one of 71,72,75,76,79) while the other scales are devoid of the white scale identified in the hindwing area has Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

288 Current status and challenges for conservation and sustainable use of biodiversity broad adwing (lower lamina) similar to the this area is that scale no: 149 (Plate B7) possess abwing(upper lamina)(Plate B5: 115). Several grey small starting from two or more buds (Plate B6: shaded scales have also been observed in this 131,132,133,134,135,139,140,141,142; Plate B7: area(Plate B5: 104,107,109,111,114; Plate B6: 144, 145, 146, 147, 148, 149, 150, 154, 128,129,130). The dimensions of white scales range 156,157,159) while the unstructured scales have between 82.74µ-106.38µ in length and 59.1µ-74.86µ broad and flat abwing. Several scales present in the in width. The close examination on the underside underside wing area is observed in the upperside too. (ventral) wing area shows long and short scales as The upperside areas have richer scales than the well as structured and unstructured scales. In this underside transverse straight lines at the lower area, it is composed of several black colored, brown lamina (adwing). The scale no: 132 (Plate B6) has colored and white colored scales. The scales appears flat upper and lower lamina. The dimensions of the to be paler than the upperside(dorsal) wing area. The underside scales ranges between 82.74µ-98.5µ in structured scale has dentations. Another finding in length and 59.1µ-74.86µ in width Table 1. The no: of scales of ‘Plain tiger’ in the respective wing area. Wing area Color Number of scales Upperside forewing area Brown 27 Upperside Forewing area Black 22 Upperside forewing area White 12 Upperside hindwing area Brown 18 Upperside hindwing area Black 24 Upperside hindwing area White 27 29 Underside area - 159 Total scales Fig. 1 The number of wing scales in ‘Plain tiger’ on various colored area Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

A preview on the wing scales of danaus chrysippus using light microscopy 289 PLATE B1 High Power View (40x) Upperside Wing Scales Forewing area 1. Brown 1 2 3 4 5 6 Length:106.38µ Length:102.44µ Length:98.5µ Length:106.38µ Length:98.5µ Length:94.56µ Width:90.62µ Width:82.74µ Width:66.98µ Width:78.8µ Width:82.74µ Width:78.8µ 7 8 9 10 11 12 Length:90.62µ Length:90.62µ Length:98.5µ Length:106.38µ Length:106.38µ Length:94.56µ Width:74.86µ Width:70.92µ Width:66.98µ Width:63.04µ Width:74.86µ Width:66.98µ 13 14 15 16 17 18 Length:102.44µ Length:110.32µ Length:98.5µ Length:106.38µ Length:98.5µ Length:98.5µ Width:70.92µ Width:74.86µ Width:70.92µ Width:82.74µ Width:66.98µ Width:82.74µ 19 20 21 22 23 24 Length:94.56µ Length:90.62µ Length:94.56µ Length:98.5µ Length:102.44µ Length:78.8µ Width:74.86µ Width:70.92µ Width:70.92µ Width:74.86µ Width:82.74µ Width:63.04µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

290 Current status and challenges for conservation and sustainable use of biodiversity PLATE B2 25 26 27 Length:90.62µ Length:98.5µ Length:102.44µ Width:66.98µ Width:70.92µ Width:66.98µ 2. Black 28 29 30 31 32 33 Length:90.62µ Length:98.5µ Length:98.5µ Length:94.56µ Length:90.62µ Length:86.68µ Width:66.98µ Width:66.98µ Width:74.86µ Width:78.8µ Width:74.86µ Width:70.92µ 34 35 36 37 38 39 Length:94.56µ Length:98.5µ Length:102.44µ Length:94.56µ Length:94.56µ Length:98.5µ Width:70.92µ Width:70.92µ Width:74.86µ Width:74.86µ Width:70.92µ Width:70.92µ 40 41 42 43 44 45 Length:82.74µ Length:110.32µ Length:90.62µ Length:82.74µ Length:94.56µ Length:82.74µ Width:66.98µ Width:63.04µ Width:66.98µ Width:70.92µ Width:74.86µ Width:66.98µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

A preview on the wing scales of danaus chrysippus using light microscopy 291 PLATE B3 46 47 48 49 Length:86.68µ Length:90.62µ Length:78.8µ Length:94.56µ Width:66.98µ Width:63.04µ Width:66.98µ Width:74.86µ 3. White 50 51 52 53 54 55 Length:102.44µ Length:94.56µ Length:90.62µ Length:86.68µ Length:86.68µ Length:90.62µ Width:66.98µ Width:66.98µ Width:74.86µ Width:70.92µ Width:70.92µ Width:70.92µ 56 57 58 59 60 61 Length:94.56µ Length:78.8µ Length:102.44µ Length:78.8µ Length:90.62µ Length:82.74µ Width:78.8µ Width:59.1µ Width:70.92µ Width:66.98µ Width:66.98µ Width:70.92µ Hindwing area 4.Brown 62 63 64 65 66 67 Length:90.62µ Length:94.56µ Length:90.62µ Length:98.5µ Length:94.56µ Length:90.62µ Width:66.98µ Width:70.92µ Width:74.86µ Width:70.92µ Width:66.98µ Width:70.92 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

292 Current status and challenges for conservation and sustainable use of biodiversity PLATE B4 68 69 70 71 72 73 Length:86.68µ Length:94.56µ Length:94.56µ Length:90.62µ Length:90.62µ Length:94.56µ Width:66.98µ Width:66.98µ Width:70.92µ Width:74.86µ Width:74.86µ Width:70.92µ 74 75 76 77 78 79 Length:90.62µ Length:98.5µ Length:86.68µ Length:90.62µ Length:94.56µ Length:90.62µ Width:66.98µ Width:70.92µ Width:70.92µ Width:66.98µ Width:66.98µ Width:74.86µ 5. Black 80 81 82 83 84 85 Length:98.5µ Length:90.62µ Length:90.62µ Length:86.68µ Length:86.68µ Length:86.68µ Width:63.04µ Width:70.92µ Width:70.92µ Width:66.98µ Width:70.92µ Width:66.98µ 86 87 88 89 90 91 Length:90.62µ Length:90.62µ Length:90.62µ Length:94.56µ Length:90.62µ Length:90.62µ Width:66.98µ Width:74.86µ Width:74.86µ Width:70.92µ Width:70.92µ Width:70.92µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

A preview on the wing scales of danaus chrysippus using light microscopy 293 PLATE B5 92 93 94 95 96 97 Length:94.56µ Length:90.62µ Length:90.62µ Length:98.5µ Length:86.68µ Length:90.62µ Width:66.98µ Width:63.04µ Width:74.86µ Width:63.04µ Width:66.98µ Width:66.98µ 98 99 100 101 102 103 Length:86.68µ Length:86.68µ Length:94.56µ Length:90.62µ Length:86.68µ Length:90.62µ Width:70.92µ Width:70.92µ Width:63.04µ Width:70.92µ Width:74.86µ Width:66.98µ 6. White 104 105 106 107 108 109 Length:90.62µ Length:90.62µ Length:86.68µ Length:86.68µ Length:90.62µ Length:106.38µ Width:66.98µ Width:74.86µ Width:70.92 Width:74.86 Width:66.98µ Width:63.04µ 110 111 112 113 114 115 Length:94.56µ Length:90.62µ Length:90.62µ Length:90.62µ Length:90.62µ Length:86.68µ Width:70.92µ Width:70.92µ Width:63.04µ Width:66.98µ Width:70.92µ Width:70.92µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

294 Current status and challenges for conservation and sustainable use of biodiversity PLATE B6 116 117 118 119 120 121 Length:94.56µ Length:90.62µ Length:94.56µ Length:90.62µ Length:90.62µ Length:86.68µ Width:63.04µ Width:70.92µ Width:70.92µ Width:70.92µ Width:70.92µ Width:66.98µ 122 123 124 125 126 127 Length:86.68µ Length:82.74µ Length:90.62µ Length:90.62µ Length:90.62µ Length:90.62µ Width:66.98µ Width:63.04µ Width:70.92µ Width:74.86µ Width:70.92µ Width:66.98µ 128 129 130 Length:86.68µ Length:90.62µ Length:94.56µ Width:70.92µ Width:66.98µ Width:59.1µ Underside Wing Scales 131 132 133 134 135 136 Length:82.74µ Length:98.5µ Length:90.62µ Length:86.68µ Length:86.68µ Length:90.62µ Width:63.04µ Width:59.1µ Width:70.92µ Width:66.98µ Width:70.92µ Width:63.04µ 137 138 139 140 141 142 Length:86.68µ Length:82.74µ Length:78.8µ Length:86.68µ Length:90.62µ Length:90.62µ Width:66.98µ Width:66.98µ Width:70.92µ Width:66.98µ Width:66.98µ Width:70.92µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

A preview on the wing scales of danaus chrysippus using light microscopy 295 PLATE B7 143 144 145 146 147 148 Length:90.62µ Length:90.62µ Length:90.62µ Length:86.68µ Length:86.68µ Length:82.74µ Width:66.98µ Width:66.98µ Width:70.92µ Width:70.92µ Width:74.86µ Width:66.98µ 149 150 151 152 153 154 Length:86.68µ Length:90.62µ Length:90.62µ Length:94.56µ Length:82.74µ Length:90.62µ Width:66.98µ Width:70.92µ Width:74.86µ Width:66.98µ Width:63.04µ Width:66.98µ 155 156 157 158 159 Length:90.62µ Length:90.62µ Length:90.62µ Length:94.56µ Length:90.62µ Width:70.92µ Width:70.92µ Width:70.92µ Width:66.98µ Width:74.86µ Discussion Müllerian mimicry – whichever the case – the evolutionary convergence of morphological The conspicuous appearance of Danaus chrysippus characters between the models and mimics seems to with a black apex and white subapical spots on the have been greatly assisted by chemical elements forewing in blight tawny-orange background including wing pigments and subject, concomitantly, coloration, likely as a typical model for various to both natural and sexual selections. mimicry species(Smith, 1973). Its wing scale pattern are linked either with Batesian or Müllerian mimicry Acknowledgement species to form a “mimicry ring”(Nishida, 2017). The authors are grateful to the Principal, Sree Conclusion Narayana College, Kollam for providing facilities to completing the work. The nature of aposematic wing coloration in Danaus chrysippusis for Müllerian mimics. Batesian or References Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

296 Current status and challenges for conservation and sustainable use of biodiversity Kinoshita,S., Yoshioka, S. and Miyazaki, J. 2008. Physics of structural colors. Rep.Prog.Phys.,71, 076401 Huang, J.Y., Wang, X.D. and Wang, Z.L. 2006. Controlled Replication of Butterfly Wings for Achieving Tunable Photonic Properties. Nano Lett., 6 (10): 2325-2331. Nishida, R. 2017. Chemical Ecology of Poisonous Butterflies: Model or Mimic? A Paradox of Sexual Dimorphisms in Müllerian Mimicry. In: Sekimura T., Nijhout H. (eds) Diversity and Evolution of Butterfly Wing Patterns. Springer, Singapore. https://doi.org/10.1007/978-981-10-4956-9-11 Smith, D. 1973. Batesian mimicry between Danaus chrysippus and Hypolimnas misippus (Lepidoptera) in Tanzania. Nature, 242:129–131 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 |pp.285-296 © Principal, Sree Narayana College, Kollam, Kerala, India

297 Currt status and challenges for conservation and sustainable use of biodiversity ISBN 978-93-5396-871-7 CHAPTER 43 AN EXPLORATION ON THE WING SCALE PATTERN OF EUPLOEA CORE Amina Thaj, Munisha Murali S* and Sheeba S PG and Research Department of Zoology Sree Narayana College, Kollam, Kerala, India *Correspondence E-mail: [email protected] ABSTRACT Euploea core, the Common Indian Crow is a glossy black butterfly belonging to the family Nymphalidae and Subfamily Danainae. The present study focused on the morphology of scales on the wings of Euploea core. Scrutiny on the dorsal side of the wings revealed dark brown coloured area, light brown coloured area, white coloured area and numerous other types of scales on the ventral side of the wings. A total of 141 different types of scales are found from the dorsal and ventral side of the wing. About 60 variety of single scales from dark brown region, 16 distinct types of scales from light brown region and 36 types of scales from the white spot region from the dorsal region and 29varieties of scales from the underside of the wing. Majority of the scales in the dark brown region are highly structured with dentations, very few scales lack dentations. The dimension of the scale ranges between 82.74µ-102.44µ in length and 59.1µ- 74.86µ in width. Few scales are found on the upper side of the wing. Most of the scales are structured except two scales are devoid of dentations. The dimension of the scales present in the light brown area of dorsal wing ranges between 86.68µ-98.5µ in length and 59.1µ-74.86µ in thickness. The white scales on the dorsal side of the wing contain both structural as well as unstructural scales. The white scales are so striking and there dimension ranges between 78.8µ-94.56µ in length and 55.16µ-70.92µ in thickness. On the ventral side of the wing 29 scales were examined and it contains brown colored scales together with white colored scales. An underside wing contains scales similar to the upper side wing scales. The overall dimensions of the wing scale ranges between 78.8µ-98.5µ in length and 59.1µ-74.86µ in thickness. These variations can have an influence in the flight of butterfly and also provides a bright coloration to the butterfly wings. Key words: Wing, Scale, Pattern, Euploea core Introduction of nonspecific individuals recommends a correlation between wing colours and Butterflies have the maximum noticeable and spectral sensitivity of their eyes(Imafuku, diverse wing colours of all insects. Butterflies 2013). are generally strikingly coloured, due to light reflected by the wing scales, which are Materials and Methods arranged on the wing surface like shingles on a roof (Giraldo, 2008). This colouration plays The butterflies of Euploea core was collected a major role in interspecific communication, from the premises of Sree Narayana College, such as aposematism (Benson, 1972), Kollam (8˚52̍55̎ N 76˚36̍4̎ E) by using mimicry (Uésugi, 1996) and as an isolating handheld insect net. Dark brown, light brown mechanism (Meyer-Rochow, 1991).The fact and white colours were observed from the that males and females react to wing colours wings.The scales are removed from the wing surface as per the standard method of Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

298 Current status and challenges for conservation and sustainable use of biodiversity Grodnitsky and Kozlov (1991). Scales were scales were studied from the underside of the detached from each region of wing separately wing (Table 1; Fig. 1). that shows varied colors into a glass slide. A drop of xylene was used for fixation of scales Firstly, the dark brown region of the and the samples were studied under the light upper(dorsal)side of the wing are closely microscope. The measurements of the scales examined through a light microscope at a were calculated using micrometry. resolution of 40x. Bright and dark brown Microscopic photography was used for taking colored scales are identified (Plate C1: 1-24; the photomicrographs of the prepared scale Plate C2: 25-54; Plate C3: 55-60). The sample for result analysis. anatomy of scales shows an upper lamina (abwing), a lower lamina (adwing) and a Results stalk. Here the majority of the scales are highly structured. Dentations of the scales Upper side dark brown and broadly paler were seen to be pointed. However, in this along terminal margins. Fore and hindwing area the scales are long and narrow (Plate C1: with sub terminal and terminal series of white 2,8,10,11,13,14,15,19,22; Plate C2: spots. On forewing the former more or less 25,29,30,50,52,53,54; Plate C3: 56,57,58). oval, curved inwards opposite to apex, the The abwing is more or less broader while the latter series often incomplete, not reaching adwing is narrow. Some scales are short and apex, the spots smaller, often there is a small flat, they possess more than two dentations costal spot and very rarely a spot in apex of (Plate C1: 1,3,4,6,9,12,17; Plate C2: 49,51). cell and one or more distal spots. On the hind wing the inner series of spots are elongate, Very often, several brown colored scales are the outer conical. Underside similar, but paler than the ground color (Plate C1: 18, 21, ground colour more uniform. Cell, costal and 24; Plate C2: 33, 36, 38, 40). However the distal spots on both fore and hind wing nearly scale no: 24(Plate C1) and scale no: 33 (Plate always present. Fore wing possess triangular C2) are devoid of dentations. The former is shape while hind wing possess nearly round larger than the latter. A few odd white scales shape. Wingtspan of fore wing ranges from are also been observed in this area (Plate 2-5 cm while that of hindwing ranges from 2- C1:16; Plate C2: 26, 27, 35, 42; Plate C3: 3 cm (Plate C). 60). Among the above scales, scale no: 60 (Plate C3) is devoid of dentations. Identical The scales were isolated from the wing scales having the similar length and width is substrate of ‘Euploea core’ and studied. The identified but they often differ from each areas under observation include the dark other due to the change in the intensity of brown colored area, light brown colored area colour(Plate C1:10,11). The scale no: 10 is and white colored region on the upper side more darker than the latter (Plate C2: 38, 39, (dorsal) and numerous scales were also 40, 41) also possess similar dimension but studied from the underside(ventral) region of they differ in the gradient of color. the wings. Another finding of the study is that, the A total of 141 different scales were studied scales having the similar dimension vary with from the upper and underside of the wing. the change in their abwing structure (Plate About 60 varieties of scales were studied C1: 13,14). Here the dimensions of the scale from the dark brown region, 16 varieties of range between 82.74µ-102.44µ in length and scales from the light brown region and 36 59.1µ- 74.86µ in width. varieties from the white spot region that constitute the upper side and 29 varieties of Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

Current status and challenges for conservation and sustainable use of biodiversity 299 Study on the light brown colored region of color. Lastly, the similar area from the the upperside (dorsal) surface of the wing ventral (underside) wing is also studied. shows only a few scales. Majority of the scales are structured (Plate C3: In the examining area, 29 scales were studied 61,62,63,64,65,67,68,69,70,71,72,73,74,75) and it contains brown colored scales together except two scales which are devoid of with white colored scales which appear to be dendation (Plate C3: 66,76). Here the scales more paler than the upper side ground color. are more or less paler than the ground color. It contains scales with structured and An odd scale is observed which is more paler unstructured upper lamina. Laminar than other scales(Plate C3: 68).Long scales characteristics of these scales contribute to with flat abwing and narrow adwing are the normal functioning of the scales. The identified(Plate C3: 61,63,64,71,73,74) while scales that lack dentations are scales the other scales appear to be flat and 115,118,119,123,129,131 (Plate C5) and short(Plate C3: 62,67,68,69,70,72,75). The scale no:139 (Plate C6) . dimensions of the scales present in the light brown area of dorsal wing ranges between Among the above mentioned scale, three 86.68µ-98.5µ in length and 59.1µ-74.86µ in variety of scales are short and flat (Plate C5: thickness. 115,131; Plate C6: 139) while the others are long with abwing broader and adwing narrow On focusing the white spot region of dorsal (Plate C5: 118,119,123,129). wing shows several white colored scales. Basically, the white spotted region contains Under side wings contains scales similar to both structural as well as unstructural scales. the upper side wing scales (Plate C5: The scales are short and broad (Plate C4: 110,124). An odd dark colored scale is 77,78,79,81,82,83,84,85,86,87,88,89,93,101, obtained in which its color is similar to the 110) except the scales 90,92,94 Plate C4). ground color. The overall dimensions of the Here the dentations of the abwing are not wing scale ranges between 78.8µ-98.5µ in pointed. The white scales are so striking and length and 59.1µ-74.86µ in thickness. The there dimensions ranges between 78.8µ- dimensions of the scales slightly vary from 94.56µ in length and 55.16µ-70.92µ in one another. These variations can have an thickness. Scale no: 108 (Plate C5) shows influence in the flight of butterfly. two pale overlapping scales producing bright Overlapping of these colored scales in the wing substrate results in the bright coloration of butterfly wing. Table 1. The number of scales of ‘Common Indian Crow’ in the respective wing area. Wing area Color No of scales Upperside(dorsal) Dark brown 60 Upperside(dorsal) Light brown 16 Upperside(dorsal) 36 Underside(ventral) White 29 - 141 Total Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

300 Currt status and challenges for conservation and sustainable use of biodiversity Fig. 1 The number of wing scales in ‘Common Indian Crow’ on various colored area PLATE C Euploea core (Common Indian Crow) Brown White Low power view (10x) Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

Current status and challenges for conservation and sustainable use of biodiversity 301 PLATE C1 High Power View (40x) Upperside Wing Scales 1.Dark brown 1 2 3 4 56 Length:90.62µ Length:98.5µ Length:94.56µ Length:82.74µ Length:94.56µ Length:90.62µ Width:63.04µ Width:59.1µ Width:70.92µ Width:66.98µ Width:66.98µ Width:66.98µ 7 8 9 10 11 12 Length:82.74µ Length:98.5µ Length:94.56µ Length:98.5µ Length:98.5µ Length:90.62µ Width:63.04µ Width:63.04µ Width:70.92µ Width:59.1µ Width:59.1µ Width:66.98µ 13 14 15 16 17 18 Length:98.5µ Length:98.5µ Length:102.44µ Length:94.56µ Length:90.62µ Length:94.56µ Width:59.1µ Width:59.1µ Width:63.04µ Width:66.98µ Width:66.98µ Width:63.04µ 19 20 21 22 23 24 Length:90.62µ Length:86.68µ Length:98.5µ Length:102.44µ Length:94.56µ Length:90.62µ Width:59.1µ Width:63.04µ Width:66.98µ Width:66.98µ Width:70.92µ Width:74.86µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

302 Current status and challenges for conservation and sustainable use of biodiversity PLATE C2 25 26 27 28 29 30 Length:102.44µ Length:94.56µ Length:98.5µ Length:90.62µ Length:90.62µ Length:90.62µ Width:63.04µ Width:66.98µ Width:63.04µ Width:70.92µ Width:66.98µ Width:63.04µ 31 32 33 34 35 36 Length:86.68µ Length:94.56µ Length:90.62µ Length:94.56µ Length:90.62µ Length:94.56µ Width:63.04µ Width:70.92µ Width:66.98µ Width:66.98µ Width:70.92µ Width:74.86µ 37 38 39 40 41 42 Length:86.68µ Length:90.62µ Length:90.62µ Length:94.56µ Length:94.56µ Length:94.56µ Width:59.1µ Width:63.04µ Width:63.04µ Width:70.92µ Width:70.92µ Width:66.98µ 43 44 45 46 47 48 Length:90.62µ Length:94.56µ Length:90.62µ Length:90.62µ Length:90.62µ Length:90.62µ Width:63.04µ Width:66.98µ Width:63.04µ Width:66.98µ Width:63.04µ Width:66.98µ 49 50 51 52 53 54 Length:82.74µ Length:90.62µ Length:90.62µ Length:94.56µ Length:94.56µ Length:98.5µ Width:66.98µ Width:63.04µ Width:70.92µ Width:63.04µ Width:63.04µ Width:59.1µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

Current status and challenges for conservation and sustainable use of biodiversity 303 PLATE C3 . 55 56 57 58 59 60 Length:90.62µ Length:94.56µ Length:94.56µ Length:94.56µ Length:90.62µ Length:94.56µ Width:63.04µ Width:63.04µ Width:63.04µ Width:63.04µ Width:66.98µ Width:74.86µ 2. Light Brown 61 62 63 64 65 66 Length:86.68µ Length:90.62µ Length:94.56µ Length:90.62µ Length:90.62µ Length:90.62µ Width:63.04µ Width:66.98µ Width:63.04µ Width:63.04µ Width:70.92µ Width:66.98µ 67 68 69 70 71 72 Length:90.62µ Length:94.56µ Length:90.62µ Length:94.56µ Length:98.5µ Length:90.62µ Width:70.92µ Width:74.86µ Width:70.92µ Width:74.86µ Width:63.04µ Width:66.98µ 73 74 75 76 Length:86.68µ Length:94.56µ Length:90.62µ Length:94.56µ Width:59.1µ Width:63.04µ Width:70.92µ Width:66.98µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

304 Current status and challenges for conservation and sustainable use of biodiversity 3. White PLATE C4 77 78 79 80 81 82 Length:78.8µ Length:78.8µ Length:82.74µ Length:78.8µ Length:86.68µ Length:86.68µ Width:66.98µ Width:63.04µ Width:63.04µ Width:63.04µ Width:70.92µ Width:66.98µ 83 84 85 86 87 88 Length:82.74µ Length:82.74µ Length:86.68µ Length:86.68µ Length:90.62µ Length:78.8µ Width:63.04µ Width:63.04µ Width:59.1µ Width:63.04µ Width:66.98µ Width:63.04µ 89 90 91 92 93 94 Length:82.74µ Length:94.56µ Length:90.62µ Length:82.74µ Length:82.74µ Length:86.68µ Width:70.92µ Width:66.98µ Width:63.04µ Width:59.1µ Width:63.04µ Width:59.1µ 95 96 97 98 99 100 Length:78.8µ Length:82.74µ Length:82.74µ Length:82.74µ Length:82.74µ Length:86.68µ Width:55.16µ Width:59.1µ Width:55.16µ Width:55.16µ Width:55.16µ Width:59.1µ 101 102 103 104 105 106 Length:82.74µ Length:78.8µ Length:90.62µ Length:86.68µ Length:90.62µ Length:86.68µ Width:63.04µ Width:59.1µ Width:70.92µ Width:59.1µ Width:59.1µ Width:59.1µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

Current status and challenges for conservation and sustainable use of biodiversity 305 PLATE C5 107 108 109 110 111 112 Length:86.68µ Length:86.68µ Length:86.68µ Length:90.62µ Length:86.68µ Length:86.68µ Width:63.04µ Width:59.1µ Width:63.04µ Width:63.04µ Width:63.04µ Width:59.1µ Underside Wing Scales 113 114 115 116 117 118 Length:90.62µ Length:86.68µ Length:78.8µ Length:86.68µ Length:90.62µ Length:90.62µ Width:70.92µ Width:66.98µ Width:63.04µ Width:59.1µ Width:63.04µ Width:66.98µ 119 120 121 122 123 124 Length:94.56µ Length:90.62µ Length:78.8µ Length:82.74µ Length:82.74µ Length:90.62µ Width:66.98µ Width:70.92µ Width:63.04µ Width:63.04µ Width:63.04µ Width:70.92µ 125 126 127 128 129 130 Length:86.68µ Length:90.62µ Length:90.62µ Length:94.56µ Length:90.62µ Length:86.68µ Width:70.92µ Width:66.98µ Width:63.04µ Width:63.04µ Width:70.92µ Width:63.04µ 131 132 133 134 135 136 Length:78.8µ Length:82.74µ Length:82.74µ Length:86.68µ Length:90.62µ Length:94.56µ Width:59.1µ Width:59.1µ Width:63.04µ Width:63.04µ Width:63.04µ Width:66.98µ Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

306 Current status and challenges for conservation and sustainable use of biodiversity PLATE C6 137 138 139 140 141 Length:90.62µ Length:90.62µ Length:86.68µ Length:94.56µ Length:98.5µ Width:63.04µ Width:74.86µ Width:70.92µ Width:63.04µ Width:66.98µ Discussion Giraldo, M. 2008. Butterfly wing scales: Pigmentation and structural properties. Adv Butterfly wing colours depend on the Insect Physiol., 38. structure and optical properties of the scales (Stavenga et al., 2004). The general structure Imafuku, M. 2013. Sexual differences in of the wing scales of the butterflies of genus spectral sensitivity and wing colouration of Euploea is independent of the colour it 13 species of Japanese Thecline butterflies produces and is of longitudinal ridges, (Lepidoptera: Lycaenidae).Eur. J. Entomol., composed of tilted scutes, cross ribs that 110(3): 435–442 connect the ridges, and trabeculae, which link the cross ribs to the membrane of the wing Meyer Rochow, V. B. 1991. Differences in scale’s upper lamina (Ruangchai, 2018). ultraviolet wing patterns in the New Zealand lycaenid butterflies Lycaenasalustius, L. Conclusion rauparahaand L.feredayi as a likely isolating mechanism. J. R. Soc. N. Z., 21: 169–177. The study of the wing scale microstructure of the butterflies in the Euploea genera indicates Ruangchai,S., PhlaiNgam,S. and that the patterns of the wing scale structure are genus-specific and that, despite showing Tungpairojwong, N. 2018. Species Diversity similar colours, the wing scales are equipped with different mechanisms exemplifying and Structure of Wing Scales of Euploea and diversity in structural coloration in nature. Papilio Butterflies from Phromlaeng, Nam Acknowledgement Nao National Park, Phetchabun Province. The authors are grateful to the Principal, Sree Narayana College, Kollam for providing Walailak Journal of Science and Technology facilities to completing the work. (WJST), 17(3), 167-180. References Stavenga, D.G., Stowe, S., Siebke, K., Zeil, J. Benson, W.W. 1972. Natural selection for and Arikawa, K. 2004. Butterfly wing Mullerian mimicry in Heliconiuserato in colours: scale beads make white pieridwings Costa Rica. Science., 176: 936–939. brighter.Proc. R. Soc. Lond. B., 271: 1577– 1584 Uesugi, K. 1996. The adaptive significance of Batesian mimicry in the swallowtail butterfly, Papiliopolytes (Insecta, Papilionidae): associative learning in a predator. Ethology, 102: 762–775. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.297-306 © Principal, Sree Narayana College, Kollam, Kerala, India

307 A look into the tiny flat plates sheathing the flimsy wings of amata passalis using light microscopy ISBN 978-93-5396-871-7 CHAPTER 44 A LOOK IN TO THE TINY FLAT PLATES SHEATHING THE FLIMSY WINGS OF AMATA PASSALIS USING LIGHT MICROSCOPY Nidhi Soman, Surya A and Sheeba S* PG and Research Department of Zoology Sree Narayana College, Kollam, Kerala, India *Correspondence E-mail: [email protected] ABSTRACT Amata passalis is a moth popularly known as sandal wood defoliator, belonging to the family Erebidae under the order lepidoptera. The framework of this study mainly concentrates on the different types of scale cells present on the varied coloured wing areas. The study has been conducted experimentally by scrapping off different coloured scales to a glass slide and fixing it with xylene and observing it under a light microscope. The dimensions of the scales were also studied by measuring it using micrometry. A total of 68 morphologically distinct types of scales were studied, which includes 48 scales of the dorsal wing and 20 scales from ventral wing. Analysis of both the dorsal and ventral wing areas revealed the presence of several white, brown and grey coloured scales. This moth species possess a wing pattern with black coloured region surfaced by white spots. Black and white scales cannot be extracted separately, as the scales seen on the white spotted region are small sized ones. Majority of the scales of the dorsal region were point edged ones and they have a length range of about110.3µm to149.7µm and width range of 59.1µm to 78.5µm. While examining the ventral wing it was observed that majority of the scales were heart shaped ones. Dimensional analysis revealed that the scales of this area have a length range of about 130µm to 145.7µm and width range of about 55.1 µm to78.8µm. The results of this work unriddled that, even though the moths are not as much attractive as other members of the order Lepidoptera including butterflies, their flimsy wings are sheathed with variety of tiny flat plates known as ‘scales’ on its surface, that are wondrously beautiful, irrespective of their sophisticated markings and patterns. Key words: Flimsy wings, Light microscopy, Micrometry, Moth, Flat plates Introduction The scales of these insects were coloured and arranged in innumerable patterns from Moths and butterflies, diagnosed as insects elusive and cryptic to the bright and showy. having scales on their wings are grouped The wings which are the most prominent together in lepidoptera, an order which forms attribute of lepidopterans including moths part of the class insecta, the dominant and butterflies are usually covered on both division of the sub-kingdom Articulata. The the veins and membranes with two layers of awe-inspiring forms and colouration of the minute socketed scales. The present study moths and butterflies caused them to be untangles the morphology of different types attracted by nature lovers. The colouration of scale cells overlaying the translucent and patterns of their wings are formed by wings of Amata passalis commonly called as thousands of microscopic tiny flat plates known as scales sheathing like tiles on a roof. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

308 Current status and challenges for conservation and sustainable use of biodiversity sandal wood defoliator belonging to the region were point edged ones while those of family Erabidae. the ventral region were heart shaped ones. The dimensional analysis pinpointed that the Materials and Methods scales present on the dorsal and ventral wing areas shows variation both in terms of length The moth species, Amata passalis has been and width. Analysis of scales on the dorsal collected from botanical garden of Sree wing area revealed that the dimension ranges Narayana College Campus, Kollam (Plate 1). from 110.3µm to149.7µm in terms of length The scales that are overlaying the wings of and 59.1µm to 78.5µm width. Majority of the moth species were taken as the experimental scales of the ventral wing has a length range sample. Several scales have been scrapped of about 130µm to 145.7µm and width range off from different coloured regions of the of about 55.1 µm to 78.8µm. wing separately and placed on a glass slide. A drop of xylene has been added to the Discussion samples that occupies the glass slide. After it has been evaporated the samples were The flimsy wings of the moths and butterflies studied under light microscope, so that the are clothed in flattened scales which are photographs of each scale can be taken actually made from modified hairs. It is these clearly and separately. Microscopic finely ridged, hollow and microscopically photography was adopted in this study, for perforated scales which gives different taking the photomicrographs of prepared colouration and patterns to the wings of this scale samples. Followed by this, the insect as they possess colour pigments. The diameters of each scale under study were Lepidopteran scale vesture of the wings measured using morphometrics. It is further serves a variety of functions. Since these preceded for result analysis. Scales are easily detachable, it helps the insects to free their wing if it is caught in a Results spider’s web (Barish, 1999). In addition to this, the scales enable development of vivid Amata passalis is a moth belonging to the or indistinct patterns which help the family Erabidae. It is often known as the organisms protect itself by concealment and defoliator of sandalwood. This moth species camouflage, mimicry and warning. The possess a wing pattern comprising of black morphology of scales has been studied by coloured region surfaced by white spots. The Downey and Allyn(1975) and they classified forewing and hind wing have a wingspan of the scales into three groups namely hair like about 5cm and 2cm respectively. A total of or piliform, blade like or lamellae and other 68 morphologically distinct types of scales variable form. In the present study only wing were studied, which includes 48 scales of the scales are focused. Many morphologically dorsal wing and 20 scales of the ventral wing. distinct scales have been observed during the Analysis of both the dorsal and ventral wing present study on sandal wood defoliator. areas revealed the presence of several white, Majority of the scales of the dorsal region brown and grey coloured scales (Plate I.1 to were point edged ones while those of the I.2). Majority of the scales of the dorsal Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

A look into the tiny flat plates sheathing the flimsy wings of amata passalis using light microscopy 309 ventral region were heart shaped ones. Scales Allyn (1975) and pointed out that the scales vary in colour, which can be due to structure observed from the wings of certain butterflies or pigmentation (Barish, 1999). Analysis of range in size from 30 µm to 80 µm length both the dorsal and ventral wing areas of and 30 µm to 500 µm width. In the present Amata passalis revealed the presence of study it was also found that the scales present several white, brown and grey coloured on the dorsal wing area has the highest scales. The dimension of the scales ranges dimension in terms of length. But in terms of from 110.3µm to 149.7µm length and width, the values are almost similar on both 55.1µm to 78.8µm width. Similar wing areas of this engrossing moth named observations were made by Downey and Amatapassalis. PLATE I Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

310 Current status and challenges for conservation and sustainable use of biodiversity Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

A look into the tiny flat plates sheathing the flimsy wings of amata passalis using light microscopy 311 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

312 Current status and challenges for conservation and sustainable use of biodiversity Conclusion References The results of this work deciphered that, even Barish, L. 1999. Mysteries of Butterfly wing. though the moths are not as much attractive The microscope, Vol47:2, 79-92pp as other members of the order Lepidoptera including butterflies, their flimsy wings are Downey, J.C and Allyn, A.C.1975. Wing sheathed with variety of tiny flat plates scale morphology and nomenclature, Bull. known as ‘scales’ on its surface, that are Allyn Mus., The Allyn museum of wondrously beautiful, irrespective of their Entomology, Sarasota, Florida, No.31, 1- elusive and cryptic markings and patterns. 32pp Acknowledgement The authors express their gratitude to The Principal, Sree Narayana College, Kollam for providing facility in the PG and Research Department of Zoology to complete the work. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.307-312 © Principal, Sree Narayana College, Kollam, Kerala, India

ISBN 978-93-5396-871-7 CHAPTER 45 ‘SCALES’ THE COLOURFUL POWDERY STUFF RESPONSIBLE FOR THE BOGGLING PATTERN AND COLOURATION IN GRAPHIUM AGAMEMNON Surya A, Nidhi Soman and Sheeba S* PG and Research Department of Zoology Sree Narayana College, Kollam, Kerala, India *Correspondence E-mail: [email protected] ABSTRACT Graphium agamemnon, also called as Green spotted triangle butterfly or tailed jay is a predominantly green and black tropical butterfly belonging to the family Papilionoidae under the order lepidoptera. The present work looks over the different types of scale sells carpeting the varied coloured wing areas of Graphium agamemnon, by brushing off different coloured scales to a glass slide and examining it under a light microscope after it has been fixed using xylene. The dimensions of the scales were also measured, with the help micrometry. A total of 59 morphologically distinct types of scales were studied, which includes 35 scales of the dorsal wing, and 24 scales of the ventral wing. For the purpose of analysis the scales are taken from both the black and green coloured portion of the dorsal wing. Majority of the scales of this region appeared to be black in colour and most of them were narrow, thin and V shaped ones. The differences among the scales were easily identified with the presence of pointed edges. Scale cells of the dorsal wing have a length range of about 86.6µm to 126µm and width range of55.1µm to 82.7µm.On analyzing the ventral wing it was found that, less diverse scales were present here and majority of the them were short and wide. Dimensional analysis revealed that the scales of this area has a length range of about 82.7µm to 102.3µm and width range of about 55.1 µm to 78.8 µm. Results of this work clarifies that it is these colourful powdery stuff known as the ‘scales’ which are responsible for the boggling pattern and colouration in Graphium agamemnon. Key words: Boggling pattern, Butterfly, Light microscopy, Micrometry, Powdery stuff Introduction scales are arranged on the wings, in the same manner as that of roof tiles. The present study Butterflies are perhaps the most colourful and unravel the morphology of different types of alluring insect known to this world. Their scale cells surfacing the wings of Graphium boggling colour pattern makes them agamemnon commonly called as Green enchanting. Sometimes, we might think of spotted triangle butterfly belonging to the the powdery stuff that gets sticks to our family Papilionoidae. It is a wondrously hands on catching a butterfly. It is these beautiful butterfly species with the dorsal differently colourful powdery scales side of both the forewing and hind wing blanketing the wings, which gives the comprising of apple green spots on black butterfly the characteristic colouration. These background. In addition to this, two green Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

314 Current status and challenges for conservation and sustainable use of biodiversity stripes are also present on the inner edge of spots. Even though the body of this butterfly dorsal hind wing. The ventral side is very appears to be black in colour, its ventral side much similar to that of the dorsal side but, it is somewhat grey in tint. A total of 59 has a browny appearance occupied by intense morphologically distinct types of scales were dark green spots. Even though the body of studied, which includes 35 scales of the this butterfly appears to be black in colour, its dorsal wing, and 24 scales of the ventral wing ventral side is somewhat grey in tint. (Plate I.1 - I.3). For the purpose of analysis the scales are taken from both the black and Materials and Methods green coloured portion of the dorsal wing. Majority of the scales of this region appeared The butterfly species, Graphium agamemnon to be black in colour and most of them were has been collected from botanical garden of narrow, thin and V shaped ones. The Sree Narayana College Campus, Kollam differences among the scales were easily (Plate 1). The scales that are present on the identified with the presence of pointed edges. wings of this butterfly species were taken as On analyzing the ventral wing it was found the experimental sample. Several scales have that, less diverse scales were present and been brushed off from different coloured majority of them were short and wide. regions of the wing separately and placed on a glass slide. A drop of xylene has been Dimensional analysis revealed that the scale added to the samples that occupies the glass cells of the dorsal wing have a length range slide. After it has been evaporated the of about 86.6µm to 126µm and width range samples were studied under light microscope, of 55.1µm to 82.7µm. In case of ventral so that the photographs of each scale can be wing, the scales of this area have a length taken clearly and separately. Microscopic range of about 82.7µm to 102.3µm and width photography was adopted in this study, for range of about 55.1 µm to 78.8 µm. In taking the photomicrographs of prepared addition to this, it was also identified that, scale samples. Followed by this, the three v shaped scales, having the highest diameters of each scale under study were dimension in terms of width among all the measured using morphometrics. It is further scales that are analyzed, were also found at preceded for result analysis. this ventral region. Results Discussion Graphium agamemnon, most often called as The amazing and eye-catching colouration of tailed jay is a swift-flying butterfly belonging the butterflies and moths are due to the to the family Papilionoidae. The dorsal side minute scales carpeting their wings. Actually of both the forewing and hind wing it is these different coloured scales which comprises of apple green spots on black give the butterflies the characteristic background. Two green coloured stripes were colouration. Studies have identified that the also present on the inner edge of dorsal hind differences that is shown by the scales in wing. The ventral side is very much similar their structure depends on their location on to that of the dorsal side but, it has a browny the wing and is independent of the colour. In appearance occupied by intense dark green addition to this, the scales enable spots, as well as some dark patches and red development of vivid or indistinct patterns Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

‘SCALES’- The colourful powdery stuff responsible for the boggling pattern … 315 which help the organisms protect itself by (Barish,1999). Analysis of dorsal wing areas concealment and camouflage, mimicry and of Graphium agamemnon revealed the warning. Since these Scales are easily presence of black coloured scales. The detachable, it helps the insects to free their dimensions of the scales range from 82.7µm wing if it is caught in a spider’s web (Barish, to 126µm length and 55.1µm to 82.7 µm 1999). The morphology of scales has been width. Similar observations were made by studied by Downey and Allyn (1975) and Downey and Allyn (1975) and pointed out they classified the scales into three groups that the scales observed from the wings of namely hair like or piliform, blade like or certain butterflies range in size from 30 µm lamellae and other variable form. In the to 80 µm length and 30 µm to 500 µm width. present study only wing scales are focused. In the present study it was also found that the Many morphologically distinct scales has scales present on the ventral wing area has been observed during the present study on the highest dimension in terms of width. Graphium agememnon. Majority of the scales of the dorsal wing were narrow, thin Conclusion and V shaped ones. The differences among the scales were easily identified with the The results of this work throws light on the presence of pointed edges. On analysing the fact that it is these colourful and sticky ventral wing it was found that, less diverse powdery stuff known as the ‘scales’ made of scales were present here and majority of them chitin, which is responsible for the boggling were short and wide. Scales vary in colour, pattern and colouration in the charismatic which can be due to structure or pigmentation butterfly, Graphium agamemnon. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

316 Current status and challenges for conservation and sustainable use of biodiversity Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

‘SCALES’- The colourful powdery stuff responsible for the boggling pattern … 317 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

318 Current status and challenges for conservation and sustainable use of biodiversity Acknowledgement The authors express their gratitude to The Principal, Sree Narayana College, Kollam for providing facility in the PG and Research Department of Zoology to complete the work. References Barish, L. 1999. Mysteries of Butterfly wing. The microscope, 47(2): 79-92pp Downey, J.C and Allyn, A.C.1975. Wing scale morphology and nomenclature, Bull. Allyn Mus., The Allyn museum of Entomology, Sarasota, Florida, 31:1-32pp Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.312-318 © Principal, Sree Narayana College, Kollam, Kerala, India

ISBN 978-93-5396-871-7 CHAPTER 46 A SWOT UP ON THE FUR-LIKE SCALES SHROUDING THE GOSSAMERY WINGS OF OLEPA RICINI USING LIGHT MICROSCOPY Nidhi Soman, Surya A and Sheeba S* PG and Research Department of Zoology Sree Narayana College, Kollam, Kerala, India *Correspondence E-mail: [email protected] ABSTRACT Olepa ricini is a moth commonly called as Darth maul moth belonging to the family Erebidae under the order lepidoptera. The present work scrutinizes the different types of scale cells overlaying the varied coloured wing areas of Olepa ricini, by dislodging different coloured scales to a glass slide and observing it under a light microscope after it has been fixed using xylene. By using micrometry, the dimensions of the scales were also measured. A total of 136 morphologically distinct types of scales were studied, which includes 70 scales of the dorsal wing, comprising of 35 scales from orange/crimson coloured region and 35 scales from brown spotted region. Among the 65 morphologically different types of scales of the ventral wing that has been studied, 41 scales were from brown spotted region and 23 scales from cream shaded region. Analysis of dorsal wing areas revealed the presence of several pink, light brown and silver coloured scales. Majority of the scales of this region were broad and large sized, and the remaining ones are long and thin. At the top of certain scales, two or more pointed edges are present, through which we could identify the differences among the scales. Scale cells of the dorsal wing have a length range of about 63.9µm to 130.04µm and width range of 90 µm to 130.04µm. On analysing the ventral wing it was found that majority of the scales were brown in colour and most of them were flat and wide. Dimensional analysis revealed that the scales of this area have a length range of about 90.6µm to 140µm and width range of about 55.1µm to 82.7 µm. The results of this work unravel that, even though the moths are not as much attractive as other members of the order Lepidoptera including butterflies, their gossamery wings are shrouded with variety of fur-like scales on its surface, that are stunningly beautiful, irrespective of their visible colour patterns. Key words: Light microscopy, Micrometry, Moth, Fur-like scales Introduction moths. They can be seen at any time of the year, with different species active in different The order Lepidoptera meaning ‘Scaly months, including mid-winter too. The winged’ includes the insects Moths and colouration and patterns of their wings are butterflies. Although moths have relatively formed by thousands of microscopic scales, dull wing colours, many species with overlaying like tiles on a roof. These scales spectacular colours and patterns also exists. can be easily dislodged as powdery dust. The Moths vary greatly in appearance as well as present study unravel the morphology of in size. The larger moths are the macro- different types of scale cells surfacing the moths and the smallest ones are the micro- wings of Olepa ricini commonly called as Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.319-326 © Principal, Sree Narayana College, Kollam, Kerala, India

320 Current status and challenges for conservation and sustainable use of biodiversity Darth maul moth belonging to the family studied, 42 scales were from brown spotted Erebidae. region and 23 scales from cream shaded region (Plate I.1 to I.4). Analysis of dorsal Materials and Methods wing areas revealed the presence of several pink, light brown and silver coloured scales. The moth species, Olepa ricini has been Majority of the scales of this region were collected from botanical garden of Sree broad and large sized, and the remaining ones Narayana College Campus, Kollam (Plate 1). are long and thin. At the top of certain scales, The scales that are present on the wings of two or more pointed edges are present, moth species were taken as the experimental through which we could identify the sample. Several scales have been scrapped differences among the scales. Majority of the off from different coloured regions of the scales present on the brown spotted region of wing separately and placed on a glass slide. the dorsal wings are two pointed edged ones. A drop of xylene has been added to the The scales that are surfacing the ventral samples that occupies the glass slide. After it wings are flat and wide and most of them has been evaporated the samples were were in brown colour. Although the analysis studied under light microscope, so that the of the ventral wing revealed the presence of photographs of each scale can be taken some variety scales, the colouration, pattern clearly and separately. Microscopic and shape of the scales of this region is photography was adopted in this study, for almost similar to that of those scales that are taking the photomicrographs of prepared surfacing the dorsal wing. scale samples. Followed by this, the diameters of each scale under study were The dimensional analysis pinpointed that the measured using morphometrics. It is further scales on the brown spotted and preceded for result analysis. orange/crimson coloured region of the dorsal side shows variation both in terms of length Results and width. At the orange/crimson coloured region, the dimension of the scales ranged Olepa ricini is a moth belonging to the from 90µm to 122.1µm length and 63µm to family Eribidae. Dorsal and ventral wing 82.7µm width. Analysis of scales on brown have distinct colours. Fore wing have fuscous spotted region of the dorsal wing revealed brown, with very numerous pale ringed black that the dimension ranges from 90.6µm to spots in the interspaces whereas hind wing is 130.04µm in terms of length and 55.1µm to orange/crimson coloured with anti medial, 82.7µm width. Scales of ventral side shown medial, post medial and marginal and more that the dimension of the scales on brown or less conjoined blotches. On the ventral spotted region ranged from 90.6µm to side of the wing, cream and brown coloured 140.2µm in length and 55.1µm to 78.5µm in patches are seen. A total of 135 scales have width. It was also found that the scales on the been studied, which includes 70 scales of the cream coloured region ranges from 90.5µm dorsal wing, comprising of 35 scales from to 122.1µm in length and 59.1µm to 82.7µm orange/crimson coloured region and 35 scales in width. from brown spotted region. Among the 65 scales of the ventral wing that has been Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.319-326 © Principal, Sree Narayana College, Kollam, Kerala, India

A swot up on the fur-like scales shrouding the gossamery wings of olepa ricini using light microscopy 321

322 Current status and challenges for conservation and sustainable use of biodiversity Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.319-326 © Principal, Sree Narayana College, Kollam, Kerala, India

A swot up on the fur-like scales shrouding the gossamery wings of olepa ricini using light microscopy 323

324 Current status and challenges for conservation and sustainable use of biodiversity Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.319-326 © Principal, Sree Narayana College, Kollam, Kerala, India

A swot up on the fur-like scales shrouding the gossamery wings of olepa ricini using light microscopy 325 Discussion detachable, it helps the insects to free their wing if it is caught in a spider’s web (Barish, The powdery stuff blanketing the diaphanous 1999). The morphology of scales has been wings of the moths are the tiny scales which studied by Downey and Allyn(1975) and they are actually made from modified hairs. It is classified the scales into three groups namely these pigmented shimmering scales which hair like or piliform, blade like or lamellae gives different colouration and patterns to the and other variable form. In the present study wings of this insect. The scales enable only wing scales are focused. Many development of vivid or indistinct patterns morphologically distinct scales have been which help the organisms protect itself by observed during the present study on Darth concealment and camouflage, mimicry and maul moth. Majority of the scales present on warning. Since these Scales are easily the dorsal wing regions were broad and large

326 Current status and challenges for conservation and sustainable use of biodiversity sized, and the remaining ones are long and References thin. At the top of certain scales, two or more pointed edges are present, through which we Barish, L. 1999. Mysteries of Butterfly wing. could identify the differences among the The microscope, 47(2):79-92 scales. On analyzing the ventral wing it was found that majority of the scales were flat Downey, J.C and Allyn, A.C.1975. Wing and wide. Scales vary in colour, which can be scale morphology and nomenclature, Bull. due to structure or pigmentation Allyn Mus., The Allyn museum of (Barish,1999). Analysis of dorsal wing areas Entomology, Sarasota, Florida, 31: 1-32 of Olepa ricini revealed the presence of several pink, light brown and silver coloured scales, whereas most of the scales present on the ventral wing region is brown in colour. The dimension of the scales ranges from 90µm to 140µm length and 55.1µm to 82.7 µm width. Similar observations were made by Downey and Allyn (1975) and pointed out that the scales observed from the wings of certain butterflies range in size from 30µm to 80µm length and 30µm to 500µm width. In the present study it was also found that the scales present on the ventral wing area has the highest dimension in terms of length. But in terms of width, the values are almost similar on both wing areas of this fascinating moth named Olepa ricini. Conclusion The results of this work unravel that, even though the moths are not as much attractive as other members of the order Lepidoptera including butterflies, their gossamery wings are shrouded with variety of fur-like scales on its surface, that are stunningly beautiful, irrespective of their visible colour patterns. Acknowledgement The authors express their gratitude to The Principal, Sree Narayana College, Kollam for providing facility in the PG and Research Department of Zoology to complete the work. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.319-326 © Principal, Sree Narayana College, Kollam, Kerala, India

A glance to aquatic entomofauna of Sasthamkotta Lake in Southern Kerala 327 CHAPTER 47 ISBN 978-93-5396-871-7 A GLANCE TO AQUATIC ENTOMOFAUNA OF SASTHAMKOTTA LAKE IN SOUTHERN KERALA Munisha Murali S*, Sheeba S and Lichu Thampi PG and Research Department of Zoology Sree Narayana College, Kollam - 691001 *Correspondence E-mail: [email protected] ABSTRACT The aquatic insects in Sasthamkotta Lake were signified by 16 families under five different insect orders. Amongst the entomofauna order Diptera was dominant and diverse with seven families. The highest mean value of Shannon-Wiener diversity index(1.886) recorded during June and low (0.6616) in March. The higher richness index (2.401) was observed in June and low (0.4809) during March. The overall dominance index of aquatic insects in Sasthamkotta Lake was 0.1886.This initial study on entomofauna indicates that lake is under slow pollution and endorses conservation steps for protection. Keywords: Aquatic insects, Sasthamkotta lake, Diptera, Diversity indices Introduction longitude. The lake has a catchment area of 934.56 hectares and surface area of 373 Aquatic insects spend a part or entire life hectares. The samples were collected cycle in water. These insects are used as randomly from six stations during the months indicators of water quality in freshwater from February 2016 to June 2016. Aquatic ecosystem. The natural and anthropogenic insects were collected using D-frame aquatic stress experienced by the water can be net(1 x 1 m area, mesh size 500 µm) and drift measured by the different tolerance level of net. The 30m expanse of reach between the these organisms. Knowledge on distribution stations was taken during the collection. The and diversity improve the understanding on samples from each station were pooled the health of the aquatic ecosystem. together and preserved in 80% alcohol. The Entomofauna play an important role in the specimens were sorted, observed and proper maintenance of the freshwater aquatic identified up to the best possible taxonomic ecosystem. Sasthamkotta Lake, the largest level using appropriate field keys (Peterson et fresh water lake in Kerala, is the sole water al., 1981; Tonapi, 1983; Morse et al., 1994; furnishing system to the Kollam city. On this Subramanian and Sivaramakrishnan, 2007). background it is necessary to note the The biodiversity indices of aquatic insects condition of the water with reference to were determined with the help of Past aquatic insects in the lake. Software. Materials and Method Result and Discussion The present investigation was carried on the A total of five different insect orders diversity of aquatic insects in Sasthamkotta belonging to 16 families were found during Lake in Kollam district in Kerala. the study. Diversity of aquatic insects Sasthamkotta Lake is located between 9°0’- collected from Sasthamkotta Lake was 9°5’ N latitude and 75°35’-76°46’ E illustrated in Table1. The orders were Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.327-330 © Principal, Sree Narayana College, Kollam, Kerala, India

328 Current status and challenges for conservation and sustainable use of biodiversity Coleoptera, Diptera, Hemiptera, Odonata and times spotted the presence of the Dytiscidae Trichoptera. Of these,seven families such as family in Coleoptera. Prevalence of Ceratopogonidae, Chironomidae, Culicidae, Hemipteran insects also suggested that the Scatopsidae and 3 unknown belong to lake was getting polluted. The number of Diptera, three families such as Carabidae, individuals of aquatic insects was found to be Dytiscidae, and one unknown family belong maximum in the month of June, which may to Coleoptera, three families such as be due to the riparian vegetation and suitable Gerridae, Hebridae and Vellidae belong to substrates. During March the abundance and Hemiptera, two such as Gomphidae and richness of the aquatic insects were very less. Libellulidae belong to Odonata and one Due to extreme drought a few species of families such as Limnocentropodidae belong insects can tolerate the habit changes and to Trichoptera. Dipterans were the dominant destruction of aquatic vegetation. Water level and trichoptera the least group in the lake. in the lake extremely decreased during The presence of Diptera indicates that water summer, due to this property shores of the bodies are organically enriched (Anjana and lakes was dried out and vegetation were also Janak, 2015). decreased abundantly. This was affected During the monsoon season most of the negatively by the aquatic insect community, pollutants were drained into the lake. Those after the introduction of the monsoon the species richness also increased. Table 1. Aquatic insects collected from Sasthamkotta Lake during the period from February to June 2016 Order Family Feb Mar Apr May Jun Coleoptera Carabidae 0 0 001 Coleoptera Dytiscidae 0 0 004 Coleoptera Unknown 0 0 010 Diptera Ceratopogonidae 0 0 101 Diptera Chironomidae 0 0 022 Diptera Scatopsidae 2 3 000 Diptera Culicidae 0 0 010 Diptera Unknown 0 0 200 Diptera Unknown 0 0 100 Diptera Unknown 2 0 000 Hemiptera Gerridae 4 5 508 Hemiptera Hebridae 0 0 003 Hemiptera Vellidae 0 0 001 Odonata Gomphidae 0 0 001 Odonata Libellulidae 0 0 847 Trichoptera Limnocentropodidae 1 0 000

A glance to aquatic entomofauna of Sasthamkotta Lake in Southern Kerala 329 Table 2. Diversity indices of aquatic insects of Sasthamkotta Lake (monthly variation) during the period from February to June 2016 Indices Feb Mar Apr May Jun Taxa_S 4 2 5 5 9 Individuals 9 8 17 8 28 Dominance_D Simpson_1-D 0.3086 0.5313 0.3287 0.3125 0.1862 Shannon_H 0.6914 0.4688 0.6713 0.6875 0.8138 Evenness_e^H/S 1.273 0.6616 1.386 1.886 Brillouin 0.8929 0.9689 1.3 0.7328 Menhinick 0.9153 0.5032 0.7337 0.8 1.539 Margalef 1.333 0.7071 1.025 0.9283 1.701 Equitability_J 1.365 0.4809 1.213 1.768 2.401 Fisher_alpha 0.9183 0.9544 1.412 1.924 0.8585 Berger-Parker 2.759 0.8559 0.8076 0.8614 4.593 0.4444 0.625 2.387 5.705 0.2857 0.4706 0.5 Table 3. Diversity indices of aquatic insects June. During the month of June not only of Sasthamkotta Lake (total variation) during great number of species present, but the the period from February to June 2016 individuals in the community are distributed more equitably among the species. Based on Indices Total the total equitability value of all months Taxa_S 17 (0.7518) it could be described that Individuals 70 equitability, or eveness of individuals, Dominance_D distributions among families in this Simpson_1-D 0.1886 community was relatively high. Shannon_H 0.8114 Evenness_e^H/S Different levels of disturbance have different Brillouin 2.13 effects on aquatic insect diversity. In order to Menhinick 0.495 preserve the biodiversity of the Sasthamkotta Margalef 1.843 Lake it was necessary to understand the Equitability_J 2.032 status of organisms. Thus the impact by Fisher_alpha 3.766 different environmental stress on functioning Berger-Parker 0.7518 of the aquatic system can be recognized. The 7.145 present study explored that the lake is getting 0.3143 polluted. On this background biomonitoring studies are constructed to protect the biodiversity by controlling pollution level. Diversity indices (monthly variation and total Since short term sampling does not cover all variation) of aquatic insects in Sasthamkotta the species which are active in different Lake are illustrated in the Table 2 and 3. The seasons of year, therefore the true number of result indicates that diversity and evenness in species estimated for the lake might be the lake was much higher in the month of higher than the presented value

330 Current status and challenges for conservation and sustainable use of biodiversity References Anjana, C. and Janak, A. 2015. Biodiversity of Freshwater Insects: A Review. The International Journal Of Engineering And Science. 4 (10):25-312 Morse, J.C., Yang, L. and Tian, L. 1994. Aquatic insects of China useful for monitoring water quality.Hohai University Press, Nanjiing People’s Republic of China, pp 569 Peterson, B.V., Mc Alpine, J. F., Shewell, G.E., Teskey, H.J., Vockeroth, J.R. and Wood, D.M. 1981. Manual of NearticDiptera. Biosystematics Research Institute Ottawa, Ontario. Vol I. pp 674. Subramanian, K.A. and Sivaramakrishnan, K.G. 2007. Aquatic Insects of India: A Field Guide. Ashoka Trust for Research in Ecology and Environment (ATREE), Bangalore, pp 62 Tonapi, G.T. 1983. Freshwater Animals of India .Oxford and IBH Publishing Co.

331 Current status and challenges for conservation and sustainable use of biodiversity ISBN 978-93-5396-871-7 CHAPTER 48 JUVENILE HORMONE MIMIC FROM MEDICINAL PLANT ANDROGRAPHIS PANICULATA (Burm f.) Bindu O* and Muraleedharan D epartment of Zoology, Sree Naryana College, Cherthala, Alapuzha, Kerala *Correspondence E-mail: [email protected] ABSTRACT Juvenile hormones (JHs) are sesquiterpenoid compounds secreted by corpora allata (CA), the cephalic ectodermal endocrine glands. A wide array of fundamental physiological processes including growth, development, metamorphosis and reproduction are known to be modulated by juvenoids or JHanalogues (JHa). From the medicinal plant species of Andrographis paniculata (Acanthaceae), JH agonists (JHa) have been isolated, identified and characterized. JH mimicking activity was tested in nymphal forms (fifth instar) cotton bug, Dysdercus cingulatus (Heteroptera: Pyrrhocoridae) that produced supernumerary sixth instar nymphs and adultoids with several morphological malformation with suboptimal dosages. Juvenoid effects of isolated JH agonist fractions on the general protein profile and induction of related vitellogenic and stress proteins were assessed in fat body by SDS-PAGE and MALDI-TOF mass spectrometric analysis. This juvenoid promises as future effective endocrine based insecticidal phytojuvenoid biomolecules. Key words: Andrographis paniculata, Dysdercus cingulatus, juvenoids. Introduction development of newer pest management strategies. Juvenile hormones (JHs) are sesquiterpenoid compounds secreted by corpora allata or One of the attempts to utilize the JH equivalent glandular tissues exclusively endocrine system as a pesticidal target was specific to insects and other selected to develop JH analogues or juvenoids. The arthropods that regulate many aspects of juvenoids first reported were farnesol and insect physiology such as metamorphosis, farnesal isolated from insects themselves. reproduction, diapause and polyphenism Later the discovery of the ‘paper factor’ (Gilbert et al., 2000; Goodman et al., 2005; from the American coniferous tree, Abies Chieka Minakuchi and Riddiford, 2006; balsamea that opened up a new research line Toyomikotaki et al., 2009). The possibility (Slama and Williams, 1966) in this direction. of disturbing the physiology of an insect by Several JH analogues (JHa) or juvenoids, interfering with its JH endocrine system was which elicit hormonal imbalance thus first recognized by Williams (1956). Since disrupting those JH influencing vital then the JH endocrine system has never functions in the insect (Muraleedharan, ceased to be a prominent target for the 1988). Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.331-338 © Principal, Sree Narayana College, Kollam, Kerala, India

332 Current status and challenges for conservation and sustainable use of biodiversity Several papers have been published on the then activated by placing in an oven for 1 hour at 110°C. Eluting solvent is prepared phytojuvenoidal properties of crude extracts using hexane and ethyl acetate at 1:2 ratio. The eluting solvent was poured on to the from different plant species (Bede et al., tank and the inner sides of the tank are covered with filter paper to saturate the 1999, 2001; Hebsy Bai and George Koshy, surrounding atmosphere with solvent vapour. It was incubated for an hour. Test samples 2004; Alvarez et al., 2007; Malarvannan et and standard cold JH (10µg) were applied as discrete spots and placed in the tank al., 2008; Baskar et al., 2009; Jeyasanker et containing the eluting solvent. When the solvent reached near the top of the plate, the al., 2010; Jagajothi and Martin, 2010, Seok plate was removed from the tank and dried in air, then placed in iodine chamber to Hee Lee et al., 2015). In this context, the identify specific spots. present study has been decided to isolate, Topical application of crude plant extracts characterise and test in detail the insecticidal Known quantities of the residue were weighed out and appropriate concentrations effect of metabolites from the plant species were prepared. Usually, 1mg of the residue was dissolved in 1ml of acetone. So that namely Andrographis paniculata each µl contained 1µg of the isolate. From such stock solutions, 5 µl, 10 µl, 15 µl, 20 (Acanthaceae). µl, 25 µl and 30 µl was topically applied on the dorsal abdominal side of the newly Materials and methods emerged fifth instar nymph using a micro liter syringe (Hamilton). All standards were The test insect: The red cotton bug treated with same dosage of cold synthetic Dysdercus cingulatus Fabr. (Heteroptera: JH III. Controls were treated with acetone. Pyrrhocoridae) used for all the bioassays. Control and treated insects were maintained in separate containers. Plant material used: Nilavepu, “King of Bitters” (Andrographis paniculata (Burm Collection of fat body f.) plant belonging to the Two days after the treatment abdomens of Kingdom: Plantae, Order:Lamiales, Family: both treated and control nymphs were cut Acanthaceae,Genus:AndrographiPreparatio open; recovered fat body removed the n of plant extract adhering hemolymph by washing in insect Ringer. The samples were homogenized in a A. paniculata whole plant (AP) was pre-chilled homogenizer in the collected, washed with water to remove dirt homogenizing medium (300µl of 0.5M Tris and sand, and shade dried and finely ground. 50 gms of powdered plant material was repeatedly extracted with 500 ml of methanol acetone mixture (1:1) solvent. This extract was then concentrated on a Rotary evaporator (Temp. 40ºC) and residue collected and was stored in a refrigerator for further use. Thin layer chromatography Silica gel was prepared in the form of slurry by using distilled water in the ratio 1:2. The adsorbent was applied on the plate, dried and Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.331-338 © Principal, Sree Narayana College, Kollam, Kerala, India

Juvenile Hormone Mimic from Medicinal Plant Andrographis Paniculata (Burm F.) 333 HCl- pH 6.8 and 9.7 ml of distilled water) Results was centrifuged at 5,000 rpm for 10 minutes at 4°C. Supernatant was mixed with equal Insect Life history volume of sample buffer. The present work is done in Dysdercus Tris Glycine SDS-PAGE cingulatus (Heteroptera: Pyrrhocoridae), commonly called as cotton stainer, reared in The electrophoretic separation of fat body the laboratory on soaked cotton seeds. Life proteins were performed by one-dimensional history of D. cingulatus consisted of egg, Sodium-Dodecyl-Sulphate Polyacrylamide five distinct nymphal instars and the adult Gel electrophoresis (SDS- PAGE) as per the stage. method of Laemmli (1970) under discontinuous and dissociating buffer Morphogenetic effect of JH mimics systems. Gel electrophoresis was done using a vertical slab gel mini- electrophoresis unit Topical application of the crude plant (BIO-RAD Powerpac) connected to the extracts from Andrographis paniculata power supply. 1mm thick resolving gel (Plate 1) with different dosages as 5 µl, 10µl, (1.5M Tris HCl pH8.8) was prepared and the 15 µl, 20 µl, 25 µl, 30 µl on newly emerged stacking gel (0.5M Tris HCl 6.8) was over fifth instar nymphs of Dysdercus cingulatus laid on the resolving gel(10%). elicited several morphogenetic effects. Lower dosages resulted in the formation of Staining and Fixation malformed adultoids. The malformed adults show deformities on the pair of hind wings Gels were stained as per the method and antenna. The hind wings of newly described by Winder et al (1977) with minor emerged adult insects became short and modification. twisted inside but the forewings remained intact. MALDI-TOF analysis Another noticeable effect was that higher Specific electrophoretic protein bands were dosages (30µl of A. paniculata) resulted in analysed by MALDI-TOF using triptic the formation of sixth instar supernumerary digestion (Sigma in-gel protein digestion nymphs, which showed both nymphal and kit). adult characters. On continued maintenance all supernumerary nymphs eventually died Protein identification without reaching the adult stage. The data obtained from MALDI-TOF were Topical treatment of cold JHIII in the analysed using software Mascot database. respective controls resulted in development of adultoids and supernumerary nymphs. Acetone treated controls resulted in normal development only. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.331-338 © Principal, Sree Narayana College, Kollam, Kerala, India

334 Current status and challenges for conservation and sustainable use of biodiversity Fat body protein profile respective control. This result shows that A.paniculata extract induced production of SDS-PAGE of fat body protein profile of 2- certain specific additional protein bands. day old treated insects showed much variation in protein bands both in quality and The selected two prominent bands with quantity compared to fat body collected from 70KDa and 60 KDa fat body proteins of the supernumerary nymphs, treated with 30 MALDI-TOF analysis showed there are two µl of A.paniculata were enough to prove the different proteins produced. Which appears significant variation in protein band patterns to be transcription elongation factor and of. D. Cingulatus. In treated bugs prominent chaperonins with approximately 70KDa and bands observed while it is very feeble in the 60 KDa. Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.331-338 © Principal, Sree Narayana College, Kollam, Kerala, India

3Ju3v3enile Hormone MimiCcufrrroemntMstaetduisciannadl PclhaanltleAnngedsrofgorracpohnissePravnaticiounlaatand(BsuusrmtaiFna.)ble use of biodivers3i3ty2 70 KDa protein GREA_STRU0 Mass: 17569 Score: 40 Expect: 48 Matches: 6 Transcription elongation factor GreA OS=Streptococcus uberis (strain ATCC BAA-854 / 0140J) GN=greA PE=3 SV=1 CH601_SYNY3Mass: 57617Score: 42 Expect: 35 Matches: 10 60 kDa chaperonin 1 OS=Synechocystis sp. (strain PCC 6803 / Kazusa) GN=groL1 PE=1 SV=4 Current Status and Challenges for Conservation and Sustainable use of Biodiversity|2020 | pp.331-338 © Principal, Sree Narayana College, Kollam, Kerala, India