CHAPTER 3

MATERIALS AND METHODS

^iosystematic studies were conducted in order to study the diversity and

distribution of butterflies species present in different ecological zones of Shiwalik hills.

Studies were also conducted to examine the taxonomically significant morphological

features and wing venation for characterizing the species/subspecies of butterflies. An

attempt was also made to study the habit and habitats of different butterfly species and

their interrelationship with flora^ Life cycle of some economically important species

was also investigated so as to analyse the difference in the food plants and their

biology.

Shiwalik hills symbolize one of the most fragile ecosystems which extend from

29''-33<' N latitude to 74°-80.5° E longitude. These hills represent the Southern most

zone of about 8-40 km width stretching for about 800 km length in the Himalaya. In the

North, these hills fall in the districts of Udhampur, Kathua and Jammu, ultimately

merging with Reasi and Poonch districts of Jammu & Kashmir. In the North-Eeast,

these hills are present in the districts of Chamba, Kangra, Bilaspur, Una, Hamirpur,

Solan and Sirmour in Himachal Pradesh, Panchkula and Yamuna Nagar in Haryana,

Dehradun, Haridwar and Udham Singh Nagar in Uttarakhand and Saharanpur in Uttar

Pradesh. Whereas, in South, these hills are bounded by districts of Gurdaspur,

Hoshiarpur, Nawanshahar and Ropar of Punjab (Table 1; Plate 1). The low lying areas

of this zone contain alluvial river terraces and gravel beds and mainly comprise of

loose boulders and pebbles of different sizes mixed with sand, silt and clay

(Anonymous, 2004).

The Shiwalik range rises from the plains of India as low hills and goes on

attaining height till it reaches an altitude of 1000 meter. These hills gradually rise from

the Indo-Gangetic plains with a gentle slope of 3-4 degrees till they touch a rugged

topography and bare steep slopes. In this tract, some hill ranges run roughly parallel to

each other for long distances and converge at places, meet and diverge again, giving

rise to small longitudinal spindle-shaped plateaus or duns between them. With torrential

rainfall during monsoon periods, undulating topography, poor vegetation cover and

coarse medium texture of sedimentary materials, the area is subject to soil erosion

hazards (Anonymous, 2004).

35

Based on weathering and erosion, two major land forms are of common

occurrence in the area besides the hills. These are peneplains comprising of gently

tolling worn out land hills and piedmont plains which are fan shaped deposits at the

foothills brought about by streams.

The Shiwalik region comprises of rocks that fall under Cenozoic era and belong

to middle Miocene, Pliocene and lower Pleistocene period. These rocks are considered

as one of the largest storehouses of mammalian remains and other vertebrate fossils.

The study of these rocks has made it possible for the geo-scientists to probe into the

mysteries of evolution of life, climate and physiography of these periods. The era

represented by these rocks is referred to as the Age of Mammals, since during this era

considerable mammalian fossils are reported. The fossils comprise of elephants,

dinosaur and primitive trilophodonts, which fed on succulent herbage and were water

loving types, which are characteristic of worm and humid low land types. The horse

Hipparian is thought to have flourished in a Savannah or swamp type of condition

prevalent in the geological history of the Shiwaliks. The fossils of antelopes suggest

prairies, steppes or desert conditions in the past remote times.

The Shiwalik range mainly comprises of soft territory sediments-sandstone, silt

stone, shale and clay, which are readily subject to mechanical disintegration. These

ranges, being in the front, face full forces of the monsoon currents and have torrential

rains. Hence wastage of mass is very common in these areas. Thus huge landslides or

scars of old landslides is usually right in the foresaid area. Major perennial Himalayan

rivers like Chenab, Ravi, Beas, Sutlej, Yamuna and Ganga drain the area. These hills

are r.lso drained by a number of crisscrossed perennials, shallow or dry nallahas, which

are locally designated as Khad or Choes.

The vegetation along the foot hills of the Shiwaliks comprises of thorny bushes

e.g. Acacia, Zizyphus. Shorea robusta assumes prominence in the shady rocks of the

Shiwaliks. Freshwater swamp forests of Bischofia javanica, Salix tetrasperma, Pyrus

pashia and Carallia brachiata are found in and around Dehradun. Major forests of

Shiwaliks are Acacia, Bauhinia, Dodonaea, mixed semi-deciduous, Sal {Shorea

robusta) and Chir (Pinus roxburghii) forests.

The Shiwaliks have warm sub-tropical type of climate. It is quite hot in summer

(45°C) and markedly cold (5°-2°C) during winters. The summer season usually starts

36

from April and lasts upto June. The intensity of high temperature is decreased by the

dust and thunderstorms, followed by light showers of rain during hot period. The rainy

season (July to September) commences with the break of the South-West monsoon and

is regarded as the most significant climatological event from agricultural point of view.

The months of October and November in which the weather generally remains dry but

the temperature falls down are the most pleasant months of the year. The cold or winter

season begins from December and ends in March.

Biosystematic and ecological studies were conducted as follows:

BIOSYSTEMATIC STUDIES ON BUTTERFLIES

^hese studies pertaining to diversity, distribution, taxonomy and wing venation of

butterflies have been carried out as: /

Diversity and Distribution

i) Collection of Butterflies

An insect net was used in order to coolect butterflies, the net was placeu over a

resting insect settled on the ground. A net consisted of a cloth bag or nylon net bag

attached to a metal ring, which holds the mouth of the open bag and a handle to which

the metal ring was attached. A ring made up of thick wire 38 cm in diameter was used.

The depth of the bag was 75 cm. Butterflies were removed gently after they became

enclosed in the bag by a rapid twist of the handle.

The collected specimens were killed with the help of killing bottle, made up of

glass jar with broad mouth and an airtight lid. In this technique, cotton soaked with

benzene solution was kept at the base of glass jar above which a filter paper of the size

of bottle diameter were kept and this technique is called as charging of the bottle. It was

charged each day during the field collections.(Usually, butterflies were killed readily

and easily by gathering the net around it, folding the wings back so that they are closed

r:^ and finally giving its thorax a light but sharp nip between the thumb and forefinger.

il) Preservation

During field surveys, the freshly collected specimens were kept in a triangular

paper envelope, which was prepared from a thin hand made or translucent oil papers.

Each envelope contains temporary label, bearing physiographic details about locality

such as latitude, altitude, longitude, temperature, humidity etc. for taxonomy or

37

systematic study with a lead pencil. Butterflies were pinned with stainless steel needle

of 38 mm length, nos. 3 and 5 for large and 20 for small specimens. Afterwards the

packets containing butterflies were kept inside the insect boxes, with thin layer of

cotton surrounding them. Paradichlorobenzene was also kept in boxes for better

preservation of lepidopterans. In laboratory, butterflies were put in a relaxing chamber,

followed by pinning perpendicularly through the middle of thorax at a point equidistant

between the bases of forewings. The wings were spread by using paper strips.

jJI hereafter, the butterflies were allowed to dry in a dessicator for 2-3 weeks depending

upon the climatic conditions. The dried specimens were transferred to air tight insect

boxes containing powdered naphthalene and already treated with benzene. A label

written with black Indian ink was fixed for each specimen. J

iii) Identification

The general maculation venation of wings, antennae and labial palpi have been

taxonomically considered as the most reliable and stable characters for the

identification viz., a viz., diagnosis of different taxa of the butterfly. Their examination

with precision is thus, an essential pre- requisite for the adequate understanding of

various species and genera.

The structure and posture of the labial palpi were studied in details. An attempt

has been made to rectify the diagnosis of different taxa by appropriately judging the

position of palpi in relation to long axis of the body, besides the scaling pattern etc.

Similarly, the ciliate and fasciculated condition of the antennae have also been re­

examined and described carefully. General morphology of head, labial palpi and

antennae, thorax and wings were also described. Regarding wings, the maculation was

studied in the usual manner, whereas, the venation ( Fig. I) has been examined from the

self prepared permanent slides, the origin, stalking and approximation of the veins etc.

was also observed. The system of nomenclature of wings was followed after the

methods of Comstoch and Needham (summarized by Comstoch, 1918) and Peile

(1937).

With the help of relevant literature Bingham (1905), Cantlie (1962), Marshall «fe

de Niceville (1890), Evans (1932,1949), Talbot (1939,1947), Wynter-Blyth (1957), the

collection were then sorted out into various superfamiles /families/genera. Tentative

identification of some species were done from description given by Marshall & de-

38

Niceville (1890), Evans (1932,1949), Wynter-Blyth (1957). Some species was

identified after their comparison with reference collection housed at Indian Agriculture

Research Institute (I.A.R.I.), New Delhi, Zoological Survey of India (Z.S.I.), Kolkatta

and Forest Research Institue (F.R.I.), Dehradun. In order to make the present findings

meaningful for different workers in this field, the history of biosystematics and ecology

of different genera and species, particularly the cases involving revalidation of old

genera, erection of new species and new combination has been discussed in detailsy

Taxonomy and Wing Venation

Before preparing the temporary mounts of the wings of all the individuals,

series of individuals representing different specimens, were randomly selected.

Reference numbers were aiiocaieu io the individuai/species. The individuals differing

in minor morphological variations were given separate number for the confirmation of

their conspecific native at a latter stage, the studying of veins by topical application

toluene or chloroform provide only a momentry clarity to the surface of the wings.

Permanent preparation of the wings were prepared lu overcome this problem. It may be

added that variation has been considered to be a reliable and good taxonomical

character for classification and identification of different butterfly species. To begin

with the process of mounting, the wings were detached from body of the insects and

were subsequently dipped in 70% alcohol for about 10-12 hours. In case of heavy

scaling on the wings, the process of descaling was further facilitating by using a camel

hair brush (no.- 0, 00, .000) after this the wings were transferred to higher grades (80%

and 90%) of ethyl alcohol for about 5 to 10 minutes in each grade and followed by

staining in alcohol cosine. After procer dehydration in each grade of alcohol and

clearing in xylene, the wings were mounted on the glass slide in Canada Balsam. In

order to draw the outline of wings of different species, a projector was used.

ECOLOGICAL STUDIES ON BUTTERFLIES

Habit, Habitats and Food Plants

Regular marked trails in 59 localities were transversed after regular intervals of

fifteen days in the mornings and evenings. All butterflies sighted on different flowers of

different plant species were collected and identified. Different plants species visited by

bitterflies during surveys were also collected and the herbarium was made on scientific

39

lines. All the plant samples were identified and got authenticated at Forest Research

Institute (F.R.I.) and Botanical Survey of India (B.S.I.) Dehradun.

Life Cycle Studies

In order to investigate life cycle, eggs were brought from the field into the

laboratory and were placed in small circular transparent containers (diameter 10 cm,

depth 4.5 cm and photo 3). Subsequently, the later larval instar was shifted to the

relatively large transparent container (12x17 cm, 15x20 cm and photos 2 and 3) along

with fresh clippings of the food plants. The last instar larvae rearing pupation were then

shifted to small sized wire mesh eclosion cages (20 x 20 x 20 cm and 30 x 30x 30 cm,

photo 3) for pupation and proper emergence of the adults. These rearing containers and

cages were lightly closed and placed in a BOD in the laboratory which was maintained

at a constant temperature (28 + 1°C and relative humidity 70 + 5% (Smart, 1985). All

these rearing boxes were carefully examined twice or thrice a day in order to make

observations on different aspects of life history such as incubation period, hatching,

larval feeding, larval defensive behaviour, parasitism, ecdysis, larval duration,

pupation, pupal duration, parasitism and eclosion etc. Due alternation was given to

hygienic conditions such as the slaring boxes were cleaned at regular intervals to

remove faecal matter and deal insect stages etc. The fresh cuttings of host plant were

provided daily to the larvae for proper growth and minimizing their mortality rate. The

gross morphology, colouration and measurements of different immature stages i.e. the

egg, the different instar larvae and the pupae were regularly recorded. The freshly

emerged adults were kept on host food plants or artificial diet consisting of 20% sugar

solution to record their longevity. Thf pairs of freshly emerged adults of each species

were also released in large flight cage (180 x 180 xI 80 cm,' photo I) to observe their

mating and oviposition behaviour in captivity.

Abbreviated titles of periodicals in the references were indicated according to

"SERIAL SOURCES BIOSIS DATA BASE TM 2002 VOLUME".

40

Map showing localities of study of butterflies from Shlwallk Hills PLATE-1

Tablel: Physiographic details of places of collection of butterflies' fauna

S.N. LOCALITIES LATITUDE (NORTH)

LONGITUDE (EAST)

HIMACHAL

1. Kangra 32°-05'-00" 76°-16'-00"

2. Guglara 32°-18*-00" 76°-16'-00"

3. Jawali 32°-18'-00" 76°-16'-00"

4. Nagrota Surian 32''-07'-00" 76°-23'-00"

5. Pong Dam 31°-40'-00" 76''-4r-00"

6. Ransar Island 3r-40'-00" 76°-4r-00"

7. Una 32°-05"-00" 76"'-16'-00" Q '^r\o r\c< r\r\i> 76 -16-00

9. Nalagarh 30°-57"-00" 76''-22'-00"

10 Bilaspur 3r-19'-00" 76''-50'-00"

11. Solan 30°-55'-00" 77''-09'-00"

12. Nahan 30°-33'-00" 77°-2r-00"

CHANDIGARH

13. Chandigarh 30M1'-00" 76°-54'-00" PANCHKULA 14. Panchkula 30M2'-00" 76''-5r-30" PUNJAB

15. Katori Bungalow 32°-27'-00" 75°-54'-00"

16. Dhar Kalain 32°-25'-00" 75°-47'-30"

17. Dhalot 32°-25'-20" 75°-54'-40"

18. Dhar Khurd 32°-24'-00" 75°-49'-00"

19. Dunera 32°-26'-30" 75°-53'-40"

20. Bar Sudal 32°-27'-00" 75°-51'-40"

21. Charmod 32°-26"-00" 75°-46'-40"

22. Thein Dam 32°-27'-30" 75°-45'-00"

23. Moren 32°-23'-40" 75M7'-40"

24. Badwain Khuh 32''-20'-00" 75°-45'-00"

25. Kokomajari 31°-15'-00" 75M5'-00"

26. Garhshankar 31°-13'-00" 76°-11'-00"

27. Hoshiarpur 3r-32'-00" 75°-57'-00"

28. Janan 3r-57"-00" 75°-37'-00"

29. Dholbaha 3r-57"-00" 75°-37'-00"

30. BGNTemple 3r -5 r -40" 75°-37'-00"

31. Salarjan Dam 3 r - 5 r ^ 0 " 75°-37'-00"

32 Bahera 3r -5r -40" 75°-37'-00"

33. Haryana Ponga 3r-35'-40" 75"'-37'-00"

34. lappa 3r-60'-00" 75''-37'-00"

35. Mahaun 31''-60'-00" 75°-37'-00"

36. Bruhi Khud 3r-57'-00" 75°-37'-00"

37. Dehrian 3r-57'-00" 75°-37'-00"

38. Janauri 0am 3r-57'-00" 75°-37'-00"

39. Ketii 3r-57'-00" 75°-37'-00"

40. Balsua 3r-57*-00" 75''-37'-00"

41. Chauhal Dam 3r-57'-00" 75''-37'-00"

42. Nawainshahar 30''-57'-00" 75°-37'-00"

43. Roop Nagar 30''-57'-00" 76°-32'-00"

44. Rehmanpur 30''-57'-00" 76''-32'-00"

45. Anandpur Sahib 3r-14'-00" 76°-30'-00"

46. Khud Batlour 30°-02'-30" 76''-30'-00"

47. Sarsa Khud 31°-03'-00" 76°-36'-00"

48. Ranjeetpura 31''-02'-30" 76°-32'-30"

49. Ladal 30''-59'-20" 76°-33'-30'"

50. Khanpur Khuian 3r-60'-00" 76°-33'-30"

51. Nangal 3r-23'-30" 76°-22'-00"

52. Mirzapur 3r-50'-30" 76°-22'-00"

53. Bahadurpur 3r-50'-30" 76°-22'-00"

54. Mangrowal 3r-50'-30" 76''-22'-00"

55. Sawara 3r-50'-30" 76°-22'-00"

56. Dumewala 31''-50'-30" 76°-22'-00"

57. Salarian Dam 31''-50'-30" 76''-22"-00"

58 Panqra 31°-50"-30" 76°-22'-00"

59 Malewala 31°-50'-30" 76°-22'-00"

Forewing

Base

3orCula

2orCulb

1 or 1A+2A

Tornal or Hind Angle

Abdominal or Inner Margin

Costa/Upper or Costal Margin

Hindwing h

8 o r S c + R i

7 or Rs

3 or Cula

2 or Culb

lb or 1A+2A la or3A

Fig. 1: General wing venation of Hindwing and Forwing in Butterfly.