In-vitro assessment of food consumption, utilization indices and losses promises of leafworm, Spodoptera litura (Fab.), on okra crop

Journal Pre-proofs Full length article In-vitro assessment of food consumption, utilization indices and losses promises of leafworm, Spodoptera litura (Fab.), on Okra crop Ahmad Nawaz, Habib Ali, Muhammad Sufyan, Muhammad Dildar Gogi, Muhammad Jalal Arif, Abid Ali, Muhammad Qasim, Waqar Islam, Muhammad Tayab, Imran Bodla, Madiha Zaynab, Khalid Ali Khan, Hamed A. Ghramh PII: DOI: Reference:

S1226-8615(19)30477-7 https://doi.org/10.1016/j.aspen.2019.10.015 ASPEN 1463

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Journal of Asia-Pacific Entomology

Received Date: Revised Date: Accepted Date:

27 July 2019 17 September 2019 25 October 2019

Please cite this article as: A. Nawaz, H. Ali, M. Sufyan, M. Dildar Gogi, M. Jalal Arif, A. Ali, M. Qasim, W. Islam, M. Tayab, I. Bodla, M. Zaynab, K. Ali Khan, H.A. Ghramh, In-vitro assessment of food consumption, utilization indices and losses promises of leafworm, Spodoptera litura (Fab.), on Okra crop, Journal of Asia-Pacific Entomology (2019), doi: https://doi.org/10.1016/j.aspen.2019.10.015

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In-vitro assessment of food consumption, utilization indices and losses promises of leafworm, Spodoptera litura (Fab.), on Okra crop

1 2 3

Ahmad Nawaz1*, †, Habib Ali2*,†, Muhammad Sufyan1, Muhammad Dildar Gogi1, Muhammad

4

Jalal Arif1, Abid Ali1, Muhammad Qasim3 , Waqar Islam4, Muhammad Tayab10, Imran Bodla6,

5

Madiha Zaynab5 , Khalid Ali Khan7,8,9, Hamed A. Ghramh7,8,9

6 7

1 Integrated

8 9

2 Department

Pest Management Laboratory, Department of Entomology, University of Agriculture, Faisalabad, Pakistan of Entomology, University of Agriculture, Faisalabad, Depalpur Campus, Okara,

Pakistan

10

3Institute

of Insect Sciences, Zhejiang University, Hangzhou 310058, P.R. China

11

4Institute

of Geography, Fujian Normal University, 350007, China

12

5College

of life Science, Fujian Agriculture and Forestry University, China

13 14

6Insect

15 16

7 Research

17 18

8 Unit

19 20

9Biology

21

10College

22

†

23 24 25

* Correspondience: A. Nawaz ([email protected]) & H. Ali ([email protected])

Biodiversity and Conservation Group, Department of Entomology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan Center for Advanced Materials Science (RCAMS), King Khalid University, P.O. Box 9004, Abha 61413, Saudi Arabia. of Bee Research and Honey Production, Faculty of Science, King Khalid University, P.O. Box 9004, Abha 61413, Saudi Arabia Department, Faculty of Science, King Khalid University, P.O. Box 9004, Abha 61413, Saudi Arabia of Crop Science, Fujian Agriculture and Forestry University, China

These authors have contributed equally to this work

26

1

Abstract

27 28

The lepidopteran insect pests have significant importance in vegetable production. The present

29

study was performed to investigate the baseline studies about the assessment of feeding and

30

consumption potential, utilization indices and losses promises of leafworm, Spodoptera litura

31

(Fab.) on Okra. The data regarding feeding potential, food utilization and consumption indices

32

as well as losses of different larval instars were recorded and subjected to appropriate statistical

33

analysis. The results showed that, in the beginning, the approximate digestibility of various

34

instars was increase, e.g. third instar (51.36%-64.03%), fourth instar (63.42%-69.45%) and

35

fifth instar (70.25%-76.10%). However, after a certain period, the digestibility was decreased

36

and efficiency to convert the ingested food into biomass varied significantly. The consumption

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index values increased with an increase in time but the consumption and growth rate was

38

declined of fourth instar larvae. The ingestion and digestion increased of third (10.01-13.06,

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8.32-11.91 mg), fourth (11.27-17.28, 10.96-14.03 mg) and fifth (12.60-19.40, 11.93-15.28 mg)

40

larval instars. The corrected weight of consumed leaves increased with a gain in body weight.

41

However, in the third instar, a decline was observed on the last day of feeding. Maximum leaf

42

area was consumed by fifth instar larvae (44.66 cm2) followed by fourth (35.41 cm2) and third

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(27.98 cm2) instars. In conclusion, all the dependent parameters, including food utilization

44

potential, consumption indices and losses were higher for fifth instar larvae than others. These

45

results emphasized the re-establishment of fundamental (economic threshold level: ETL,

46

economic injury level: EIL) integrated pest management concepts.

47

Keywords: Lepidopteran insect pests; armyworm; okra; feeding potential; ETL; EIC

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Introduction

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Okra is native to Africa, North Australia to the Pacific and South-East Asia (Kaleri et al., 2011)

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and commercially grown in many countries of the world such as Turkey, Bangladesh, Malaysia,

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Southern United States, Afghanistan, Pakistan, Brazil, Thailand, India, Ethiopia, Iran, Western

52

Africa, Yugoslavia, Japan, Myanmar and Cyprus (Purseglove, 1987; Benjawan et al., 2007;

53

Qhureshi, 2007). In the world, its production is about six million tons per year (Benchasri, 2012).

54

In 2009-2010, the area under okra crop was 0.43 million hectares, and the production stood at 4.54

55

million tons (Benchasri, 2012). India was the first country in the world, with 3.5 million tons of

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okra production from over 0.35 million hectares of land (FAOSTAT, 2008). India is the most 2

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important country in the world for okra production and shares a part of 67.1%, followed by Nigeria

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(15.4%) and Sudan (9.3%) (Varmudy, 2011). In Pakistan, okra crop is planted on an area of 15,081

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hectares with an annual production of 114,657 tons, and it stands sixth for its okra production

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(Varmudy, 2011). Okra is a edible crop and mainly grown for obtaining fruits, but other plant parts

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like leaves, petals, stems, flowers and roots are also used as medicine, bio-fuel and for food purpose

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in several regions of the world (Aziz et al., 2011). Its nutritive value is higher than tomato, cucurbit,

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and eggplant, except bitter gourd (Nonnecke, 1989). It contains proteins, carbohydrates and

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vitamin C (Lamont, 1999; Owolarafe and Shotonde, 2004; Gopalan et al., 2007; Arapitsas, 2008;

65

Dilruba et al., 2009).

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Okra is susceptible to several diseases and a wide range of insect pests (N’Guessan et al., 1992;

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Ghanem, 2003) at its various growth stages (Ek-Amnuay, 2010; Fasunwon and Banjo, 2010). One

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of the major limitations for okra production is heavy infestations of several insect pests (Pal et al.,

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2013). Avoidable losses due to pests was at about 54% (Chaudhary and Dadheeck, 1989). Insect

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pests like crickets attack at the seedling stage while the sucking pests are common during the

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vegetative stage of the crop (Fajinmi and Fajinmi, 2010). Among chewing insects, leafworm

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Spodoptera litura (Fab.) is the most destructive pest which feeds on early vegetative stages of okra

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(Kumar et al., 2010). It is a sporadic insect pest that causes 25.8%-100% losses in crops (Dhir et

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al., 1992), depending upon the crop stage and its infestation level in the field. The major crops

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attacked by armyworm are brassica, maize, cotton, flax, lucerne, rice, soya bean, tobacco, jute, tea,

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cucurbit, potato, capsicum, tomato, eggplant (Zhou et al., 2010), cauliflower, okra, cabbage,

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radish, peanut and other legumes. It feeds gregariously on leaves leaving behind the midribs

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(Ahmad et al., 2013). It is the most destructive insect pest in the Asia-Pacific region because of its

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high reproductive potential and massive infestation rates (Ahmad et al., 2013). Eggs are laid in

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clusters and are covered with the tuft of hair to protect them from more than 100 species of

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biocontrol agents (Rao et al., 1993). A single female moth can lay more than 2000 eggs in her life

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span of 6-8 days (Ahmad et al., 2013). The eggs hatch in 2-3 days, and there may be 3-4 continuous

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layers in a single batch (Waterhouse and Norris, 1987; Hill, 1975.). Although, many biological

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(Ali et al., 2016, 2108a,b,c,d,e; Arif et al., 2018; Bala et al., 2018; Qasim et al., 2018a,b) and

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biopesticidal approcehs (Ali et al., 2015; Shakeel et al., 2018) has been made recently against

86

major crop pests.

3

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But the ever-changing climatic conditions change the behavior of insects, and there is a need

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to investigate the damaging potential of major insect pests to re-establish the economic injury

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level and economic threshold levels. Therefore, the present research was planned to investigate

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the baseline studies about the assessment of feeding potential, food consumption and utilization

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indices and losses of larval instars of S. litura on okra. This study will help to analyse the

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impact of recent climatic conditions on the behavior of S. litura in Pakistan.

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M at er i als an d met hod s

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Okra cultivar and its cultivation

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The seed of hybrid variety of okra (Adventena-803) was purchased from ICI-company and used

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as a target crop. The okra crop was sown at the University of Agriculture Faisalabad as per

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recommended sowing method (line sowing) and agronomic practices. All the recommended

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agronomic practices were performed uniformly throughout the growing season of okra. However,

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no plant protection measures were practiced at least fifteen days before the picking of leaves for

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offering to armyworm larvae.

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Preparation of larval culture of Spodoptera litura

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Leaves with egg-masses of S. litura were clipped from the mung bean field while adult female

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moths were collected from light sources near the mung bean field. The collected eggs-masses were

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placed on the moistened filter paper adjusted in petri-dishes till their hatching to get larval instars.

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Whereas moths were kept captured inside the insects rearing cages having cotton swab soaked in

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adult moth diet (honey: water: yeast : 1: 9: 1) and folded strips of black colors in hanging position.

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The egg-masses on the folded strips were scratched and also placed on the moistened filter paper

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adjusted in Petri dishes till their hatching to get larval instars. The larval culture was maintained

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in the IPM laboratory at laboratory condition (28 ± 2 ºC, 65 ± 5 % R.H). The newly emerged larvae

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of third, fourth and fifth instar were kept starved for 12h and then offered okra leaves as the food

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inside the plastic boxes in the laboratory. The materials that were used in present research include

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Electrical Balance, Plastic Boxes, Iron Net, Datasheet, Leaf Area Meter.

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Methodology

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Four sets, each containing 12 plastic boxes (total 48 boxes), were cleaned and sterilized, and each

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set was denoted as one treatment. There were four treatments [third instar larvae (T1), fourth instar

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larvae (T2), fifth instar larvae (T3) and no larvae (T0)] and each treatment was replicated 12 times 4

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for each larval instar of S. litura. In this way, 48 boxes were arranged in four sets in insect rearing

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room, maintained at 28 ± 2 ºC, 65 ± 5% R.H and 16h:8h of light and dark period.

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Ten newly emerged larvae of each (third, fourth and fifth) instar were picked from laboratory

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culture and released into each of 12 boxes of set-1, set-2 and set-3, respectively. In control

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treatment (set-4), no larvae were released. After 12 h of starvation in sample size (set-1 and set3),

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the initial accumulative weight of the ten larvae each replicate of each treatment was measured by

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weighing balance, and an average weight of larvae was calculated. Fresh, cleaned and sterilized

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okra leaves were offered as food to the larvae. Before providing these leaves, the initial area and

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weight of the leaves were measured with the help of leaf area meter and weighing balance,

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respectively. After 24 h, the old leaves were replaced with the fresh leaves.

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Similarly, the area as well weight of fresh and consumed leaves were measured before and after

128

consumption, respectively. These observations were recorded daily, and the leaf area consumed

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per day per larvae was measured. The quantity of food consumed, fecal matter excreted and larval

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growth was determined based on the fresh and dry weight. In case of control treatment, only fresh

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leaves after weighing were kept under the same set of condition to determine the natural loss of

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moisture, which was used for calculating the corrected weight of consumed leaves by equitation

133

described by Ghanema (2002).

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Corrected weight of consumed leaves = (Cb / Ca) × Ta

135

Where: Cb = Initial fresh weight of leaves without larvae; Ca = Final weight of leaves without

136

larvae; Ta = Final weight of leaves with larvae after feeding

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In the rest of the treatments (set-1 to set-3), the weight of leaves before offering and after

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consumption was determined daily. Amount of food ingested was calculated by subtracting the

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weight of residual leaves from weight of leaves given as wet and dry matter. The food digested

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was calculated by subtracting the weight of fecal matter produced from the weight of food ingested.

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The samples of leaves, faeces and larvae were dried in the oven at 80ﾟC to a constant weight. The

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leftover food and faeces were recorded and removed daily. Thus, for each instar, the increase in

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fresh and dry weights of larvae, fresh and dry weights of food eaten and digested and dry weight

144

of faeces were recorded (Rath et al., 2003; Seidavi, 2009). Fresh leaf mass (L1), consumed fresh

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leaf moisture as well as weight and moisture of faeces and un-used leaves (L2) was measured

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carefully and recorded daily. The actual leaf mass-consumed was calculated using the following

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formula: 5

148

Leaf-mass consumed = [L1 × moisture (%)]–[L2 × moisture (%)]

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Various indices of food consumption and utilization were determined by the following

150

methods/formulae described by Waldbauer (1968), Rath et al. (2003) and Seidavi (2009).

151

Approximate Digestibility (AD) =

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Efficiency of conversion of ingested food to biomass (ECI) =

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Efficiency of conversion of digested food to biomass (ECD) =

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Consumption Index (CI) =

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Growth Rate (GR) =

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Where: G = Fresh weight gain of larvae; C = Fresh weight of consumed leaves; F = Feces weight

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during feeding; T = Duration of feeding period; A = Mean fresh weight of larvae during feeding

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The whole of the experiment was laid out in CRD (completely randomized design) with twelve

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replications and four treatments.

160

Statistical Analysis

161

The data regarding indices mentioned above of food consumption and utilization were transformed

162

and then analyzed using the ANOVA technique. The means were grouped and compared using

163

Tukey HSD test using suitable statistical software package.

164

Results

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Table 1 showed analysis of variance of data regarding the dependent parameters (Quantity of food

166

ingested (mg), quantity of food digested (mg), efficiency of conversion of ingested food into

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biomass (%), efficiency of conversion of digested food into biomass (%), corrected weight of

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consumed leaves (mg), approximate digestibility (%), consumption index (mg), growth rate (mg)

Dry weight of food eaten ― Dry weight of faeces produced × 100 Dry weight of food eaten Dry weight gain of larvae × 100 Dry weight of food eaten

G × 100 C―F

C T×A

G T×A

6

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and consumed leaf area (cm2)) by third, fourth and fifth instar larvae of armyworm (S. litura) after

170

24h, 48h and 72h, respectively. Analysis of variance parameters reveal that dependent variable

171

(Quantity of food ingested (mg), quantity of food digested (mg), efficiency of conversion of

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ingested food into biomass (%), efficiency of conversion of digested food into biomass (%),

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corrected weight of consumed leaves (mg), approximate digestibility (%), consumption index

174

(mg), growth rate (mg) and consumed leaf area (cm2)) varied among different treatments (third,

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fourth and fifth larval instar) as (P < 0.05).

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The fifth instar larvae showed the highest values for all the dependent parameters (Quantity of

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food ingested (mg), quantity of food digested (mg), efficiency of conversion of ingested food into

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biomass (%), efficiency of conversion of digested food into biomass (%), corrected weight of

179

consumed leaves (mg), approximate digestibility (%), consumption index (mg), growth rate (mg)

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and consumed leaf area (cm2)) followed by fourth and third instar larvae of S.litura (Fig 1-8).

181

Discussion

182

As the results showed that approximate digestibility for the fourth instar larvae of S. litura

183

fluctuates between 63.42-69.45%. First, it increases from 66.84%-69.45% and then decreases to

184

63.42%. These results are partially consistent with the findings of Rashwan (2013) who worked

185

on cotton leafworm S. littoralis and observed a decrease in approximate digestibility (control) with

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an increase in larval age. This small difference may be due to the change of species or in the

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composition of food as they were reared on leaves of castor bean. Evans (1939) reported that the

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reasons for a decrease in approximate digestibility could be described as the insects are small

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individuals which cut off tiny parts of food and give a large surface for the process of digestion,

190

and as they grow older, the nature of the food chosen by them also varies. The results of the present

191

study indicated that efficiency to convert the ingested food into biomass (40.65%-52.42%) and

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capability to convert the digested food into biomass (39.77%-53.95%) values increased with an

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increase in time.

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In fifth instar larvae of S. litura the ingestion, digestion, consumption index and growth rate

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increases from 12.60-19.40 mg, 11.93-15.28 mg, 2.42-2.56 %, and 0.71-0.89%, respectively.

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These outcomes are in agreement with the findings of Rath (2010) who worked on nutritional

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parameters of Antheraea mylitta larvae by offering them fresh leaves of Tilia tomentosa. 7

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Nutritional indices like the relative consumption rate, ingestion, relative growth rate, digestion,

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and gain in body weight increased significantly with an increase in the number of diets per day,

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but approximate digestibility and efficiency in converting the ingested food into biomass declined,

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while ability in converting the digested food into biomass did not change.

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Approximate digestibility of third (51.36%-64.03%), fourth (63.42%-69.45%) and fifth (70.25%-

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76.10%) instar larvae first increased and then decreased, while conversion efficiency of ingested

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food into bio-mass varies significantly between 33.85-41.59%, 40.65-52.42%, 51.21-60.28% for

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third, fourth and fifth instar of armyworm, respectively. However, efficiency conversion of

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digested food into bio-mass values rose between ranges of 34.26-41.84%, 39.77-53.95%, 45.23-

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61.75% for third, fourth and fifth instar of S. litura, correspondingly. These results are in line with

208

the research outcome of Kumar and Ballal (1992) who determined the impact of parasitization by

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Hyposoter didymator on food utilization of S. litura and found that feeding potential, faeces

210

produced and weight gained were significantly less and AD was higher in infected larvae, while

211

ECI and ECD was highest in unparasitized larvae of armyworm. The outcome mentioned above

212

of present research is also consistent with the results of Reynolds et al. (1985), who found that

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approximate digestibility was about 60% while EC1 and ECD were both very high. EC1 was 43%

214

and ECD was 72% in fifth instar larvae of Manduca sexta (Lepidoptera: Sphingidae). However,

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the results were found in controversy with the findings of Teimouri et al. (2015) who determined

216

the relative consumption rate to be 5.36, 11.10 and 10.631 (mg/mg/day) on artificial diet, Akbari

217

and Kalequchi cultivars, respectively. While, according to our results, its values are between 0.84-

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0.87 mg, 1.14-1.99 mg and 2.42-2.56 mg for third, fourth and fifth larval instars of S. litura,

219

correspondingly. Carob moth Ectomyelois ceratoniae (Zeller) (Lepidoptera: Pyralidae) larvae

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reared on Akbari cultivar showed the highest ECD (5.64 ± 0.43). The highest amount of ECI was

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obtained on artificial diet, but approximate digestibility (AD) was the lowest on this diet. These

222

differences may be due to the difference between two species of insects as they have different size,

223

and their metabolism is also different from each other. It might be due to the difference in the

224

composition, nature and type of food used by Teimouri et al. (2015).

225

These results are partially similar to the findings of Rath (2010) who found that dry matter

226

ingested, digested, ECD and biomass gain increased with the increase of larval development of A.

227

mylitta. A similar trend was observed in S. litura, where ingestion (10.01-19.40 mg/72 h), digestion 8

228

(8.32-15.28 mg/72 h) and ECD (34.26%-61.75%) increased with the increase of larval

229

development. While the relative consumption rate and growth rate was declined, and this trend

230

was observed only in the fourth instar larvae of S. litura. This might be due to the difference in

231

size as the larger insects consume less energy for their growth as compared to the smaller insects,

232

despite high growth rate (Reynolds et al., 1985; Rath et al., 2003).

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Hemati el al. (2012) checked the effect of different plant species on nutritional indices of H.

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armigera. It was found that third larval instar developed on potato indicated the highest amount of

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ECD and ECI (50.80 ± 0.11% and 13.63 ± 0.02%, respectively), and the same trend of ECD and

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ECI (41.84 ± 0.06% and 41.59 ± 0.06%, respectively) was observed in the current study. They

237

reported that AD of the fourth instar larvae was highest (92.65%) on chickpea (Azad) and least

238

(57.14%) on potato (Agria). Likewise, we also found maximum and minimum AD value (69.45 ±

239

0.09 % and 63.42 ± 0.07%, respectively) for the fourth instar of S. litura. They reported that fifth

240

larval instar reared on tomato and white kidney bean showed the great amount of feeding (3.71

241

mg) and relative consumption rate (1.62 mg), respectively. However, the result of present study

242

indicated that consumption index and growth rate values of fifth larval instar of S. litura were 2.56

243

± 0.03 mg and 0.89 ± 0.03 mg, respectively.

244

Maximum leaf area was consumed by fifth instar larvae of armyworm (44.66 cm2) followed by

245

fourth (35.41 cm2) and third (27.98 cm2) instar larvae. The findings of the present work are

246

consistent with the results of Dhir et al. (1992) who determined the losses in groundnut caused by

247

the tobacco caterpillar, S. litura and found that one larva/plant consumed about 54.7% of leaf area.

248

At flowering and pegging stage, single larva per plant consumed the 49.1% and 38.8% of leaf area

249

and reduced the yield upto19% and 5.7%, respectively.

250

Conclusion

251

It was concluded that maximum feeding potential (leaf-area/weight/mass consumed per larvae),

252

food utilization and consumption indices (consumption index, conversion efficiency of ingested

253

food into bio-mass, corrected weight of consumed leaves, growth rate, conversion efficiency of

254

digested food into bio-mass, consumed area of leaf, approximate digestibility, quantity of food

255

ingested and quantity of digested diet) and losses promises (% dry-weight and wet-weight loss)

256

were observed in T3 (fifth larval instar) followed by T2 (fourth larval instar) and T1 (third larval 9

257

instar). This is due to the fifth instar larvae were larger, their body metabolism was fast that is why

258

they consumed more food. Ultimately, digestion was also significant, efficiency to covert the

259

quantity of food digested into bio-mass, consumption, growth rate and capability to covert the

260

quantity of food ingested into bio-mass values were also higher. So, by managing this pest we can

261

reduce the losses as this pest can cause 25.8%-100% losses in different crops, especially in

262

vegetables.

263

Conflict of Interest

264

All authors declare no conflict of interests

265

Acknowledgements

266

We are profoundly grateful to Higher Education Commission (HEC) Pakistan who provided the

267

financial support to this project. We also acknowledge the support of the King Khalid University

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(RCAMS/KKU/010-19) Research Center for Advanced Materials Science (RCAMS) at King

269

Khalid University, Kingdom of Saudi Arabia.

270

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Conflict of Interest

405

All authors declare no conflict of interests

Dependent Parameters

Quantity of food ingested (mg) Quantity of food digested (mg) Efficiency of conversion of ingested food to biomass (%) Efficiency of conversion of digested food to biomass (%) Corrected weight of consumed leaves (mg) Approximate Digestibility (%) Consumption index (mg) Growth rate (mg) Consumed leaf area (cm2)

DF

24h

48h

72h

F value

P value

F value

P value

F value

P value

2a/33b 2a/33b 2a/33b

12.8 12.3 20.7

0.01* 0.00* 0.00*

8.96 7.49 45.6

0.00* 0.03* 0.00*

1.00 1.30 1.68

0.03* 0.02* 0.02*

2a/33b

19.0

0.00*

44.1

0.00*

1.72

0.01*

2a/33b

3.25

0.03*

0.19

0.04*

5.34

0.00*

2a/33b 2a/33b 2a/33b 2a/33b

0.66 44.4 0.02 13.5

0.02* 0.00* 0.01* 0.00*

0.25 48.1 1.65 11.6

0.00* 0.00* 0.02* 0.00*

8.37 95.2 0.58 2.35

0.01* 0.00* 0.04* 0.01*

406 407 408

Highlights

409



The lepidopteran insect pests have significant importance in vegetable production

410



All dependent parameters and losses promises of leaf worm on Okra were evaluated

411



The consumption index values increased with an increase in time

412



All dependent parameters were higher for fifth instar larvae than others.

413



These results emphasized the re-establishment of fundamental IPM.

414 415 416 417 15

418

Table 1. Analysis of variance of data regarding quantity of food ingested (mg), quantity of food

419

digested (mg), efficiency of conversion of ingested food into biomass (%), efficiency of conversion

420

of digested food into biomass (%), corrected weight of consumed leaves (mg), approximate

421

digestibility (%), consumption index (mg), growth rate (mg) and consumed leaf area (cm2) by 3rd,

422

4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours (degree of

423

freedom of replication = 2) Treatments (3rd, 4th and 5th instars of Spodoptera litura).

424 425 426 3rd instar

Quantity of food ingested (mg)

25

4th instar

19.4 C

20 15

5th instar

17.28 A 12.98 AB 13.06 A

14.12 AB 11.27 B

10.01 B

15.29 B 12.6 A

10 5 0 24 hours

48 hours Feeding durations

72hours

427 428 429

Fig. 1. Quantity of food ingested (mg) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

430

16

3rd instar

4th instar

5th instar

Quantity of food digested (mg)

18 16 14 12 10

11.93A 10.96 B

12.94B 13.16 A 10.82C

14.03B

15.28A

11.91C

8.32 C

8 6 4 2 0 24 hours

48 hours Feeding Durations

431 432 433

72hours

Fig. 2. Quantity of food digested (mg) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

434

Efficiency of conversion of ingested food to biomass (%)

435 3rd instar

4th instar

5th instar

70

60.28A

60 50 40

51.21A 40.65B 33.85C

54.08A 45.84B 36.17C

52.42B 41.59C

30 20 10 0 24 hours

48 hours Feeding Durations

72hours

436 437 438

Fig. 3. Efficiency of conversion of ingested food to biomass (%) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

439 17

Efficiency of conversion of digested food to biomass (%)

3rd instar

4th instar

5th instar

70

60.78B 61.75A 53.95A

60 50 40

41.84A 36.29AB 34.26B

47.14B

41.59C

36.17C

30 20 10 0 24 hours

48 hours Feeding Durations

72hours

440 441 442

Fig. 4. Efficiency of conversion of digested food to biomass (%) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

443 444 445 446

Corrected weight of consumed leaves (mg)

447 3rd instar

4th instar

3.5

3.05A

3

2.72A

2.5 2

5th instar

1.83C

2.04B

2.98A 2.63AB

2.55AB 2.01B

1.91B

1.5 1 0.5 0 24 hours

48 hours Feeding durations

448

18

72hours

449 450

Fig. 5. Corrected weight of consumed leaves (mg) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

451 3rd instar

4th instar

Approximate digestibility (%)

90 80 70

70.25A 66.84AB 61.67B

69.45AB 64.03B

5th instar 76.1A

60

72.68A 63.42B 51.36C

50 40 30 20 10 0 24 hours

452 453 454

48 hours Feeding durations

72hours

Fig. 6. Approximate digestibility (%) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

455 456 457 458 459

19

3rd instar

4th instar

5th instar

Consumption index (mg)

3 2.5

2.56A

2.46A

2.42A 1.99B

2

1.98B

1.5

1.14B 0.87C

0.84C

1

0.85C

0.5 0 24 hours

48 hours Feeding durations

72hours

460 461 462 463

Fig. 7. Consumption index (mg) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

464 3rd instar

4th instar

5th instar

1

0.89A

0.9 Growth rate (mg)

0.8 0.7

0.69AB0.71A 0.61B

0.78B

0.76A 0.67AB 0.64B

0.67C

0.6 0.5 0.4 0.3 0.2 0.1 0 24 hours

465 466 467

48 hours Feeding Durations

72hours

Fig. 8. Growth rate (mg) by 3rd, 4th and 5th instar larvae of armyworm (Spodoptera litura) after 24, 48 and 72 hours of feeding

468 20