Legume Research

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Research Article

Legume Research, volume 44 issue 6 (june 2021) : 667-672

Fumigant Activity of Four Plant Powders against Cowpea Weevil, Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae) in Stored Adzuki Bean

Muhammad Bayu Mario2,3, Ludji Pantja Astuti3, Jue-Liang Hsu2, Lekhnath Kafle1,*
1Department of Tropical Agriculture and International Cooperation, National Pingtung University of Science and Technology, 1 Shuefu Road, Neipu, Pingtung 912, Taiwan (ROC).
2Department of Biological Science and Technology, National Pingtung University of Science and Technology, 1 Shuefu Road, Neipu, Pingtung 912, Taiwan (ROC).
3Department of Plant Pest and Disease, Faculty of Agriculture, University of Brawijaya, Jalan Veteran, Malang 65145, Indonesia.

  • Submitted21-10-2019|

  • Accepted25-01-2021|

  • First Online 07-04-2021|

  • doi 10.18805/LR-533

Cite article:- Mario Bayu Muhammad, Astuti Pantja Ludji, Hsu Jue-Liang, Kafle Lekhnath (2021). Fumigant Activity of Four Plant Powders against Cowpea Weevil, Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae) in Stored Adzuki Bean . Legume Research. 44(6): 667-672. doi: 10.18805/LR-533.

ABSTRACT

Background: Cowpea weevil, Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae) is one of most common species pest that attack various types of beans in the storage. Plant-based insecticides are considered as good candidates as alternatives to methyl bromide and phosphine for controlling pest in storage. Plenty of reports about plant powder as grain protectant, conversely a little information on plant powder as fumigant had been reported. This study was designed to observe the effects of plant powders of clove, holy basil, lemongrass and turmeric as fumigant on the biological parameters of C. maculatus under laboratory conditions.

Methods: Experiments were conducted on plant powders against biological parameters of cowpea weevil. No choice test method was followed to assess the bioefficacy of the four natural fumigants on adult longevity, fecundity, F1 progeny, bean damage and weight loss of bean under completely randomized design (CRD).

Result: Fumigant toxicity of four plant powders (clove, holy basil, lemongrass and turmeric) varied amongst doses. Clove powder showed significantly lower adult longevity, fecundity of female, number of F1 progeny, bean damage and weight loss of bean at 1 g and 3 g dose per container. Major volatile organic compounds (VOCs) in clove powder were eugenol (48.64%) and caryophyllene (43.09%) identified by solid phase microextraction followed with gas chromatography and mass spectrometry (SPME-GC/MS). These two major constituents might explain the fumigant toxic activities of clove powder towards C. maculatus biological parameters.

INTRODUCTION

Cowpea weevil, Callosobruchus maculatus (Fabricius) (Coleoptera: Chrysomelidae) is major pest that cause serious damages to storage beans in most of tropical countries. This insect has been categorized into primary pest that adequate to penetrate and infest of undamaged grains. Furthermore, the insect being internal feeder immature stage developes inside the bean (Singh, 2017; Rees, 2004) causing weight loss, seed germination reduction, contamination, heating of commodity, quality loss and mould growth (Rees, 2004). This pest can attack many bean species, not just in storage facility but also before harvest (Prevett, 1961; Hagstrum et al., 2013).
       
Plant-based insecticides are considered as good candidates as alternatives to methyl bromide and phosphine for controlling pest in storage (Athanassiou et al., 2014). The major traits of the plant-based pesticides, viz. relatively effective, biodegradable and considerably low mammalian toxicity, made them highly desirable environmentally friendly alternatives (Athanassiou et al., 2014). As much 1,079 plants species have been described as botanical pesticide and 866 species of them possess toxicities, repellencies, physiological and deterrent effects against insects (Prakash and Rao, 1997). Plenty of natural products have been used as pest control agents, but most of them are using liquid formulations (plant extract or essential oil) as fumigant and solid formulations (powder) as contact insecticide or grain protectant (Adedire and Akinneye, 2004; Boeke et al., 2004: Hernandez-Lambraño et al., 2015). Plenty of reports about plant powder as grain protectant, conversely a little information on plant powder as fumigant had been reported (Weaver et al., 1994).
       
This study was designed to observe the effects of plant powders of clove, holy basil, lemongrass and turmeric as fumigant on the biological parameters of C. maculatus under laboratory conditions.

MATERIALS AND METHODS

Insect diet
 
Kaohsiung-9 variety of adzuki bean obtained from Pingtung and Donggang Farmer’s Association Taiwan was used for rearing and further experimentation after cold sterilization as per Heinrichs et al., (1985) and Astuti et al., (2019).
 
Insect rearing
 
Inoculative insect culture was obtained from the Taiwan Agricultural Research Institute (TARI) in Taichung, Taiwan. Adzuki bean (100 g) was put in plastic container (Ø=6.5 cm, h=5 cm), then infested with unsexed 50 adults of C. maculatus. The lid of plastic containers was covered by nylon mesh. Eggs laid were allowed to develop inside the beans and emerged adults were collected for experimental purpose. Adults infestation were removed after all adults were dead.
 
Natural fumigants oreparation
 
Four kinds of herbs were respective plant parts such as buds (clove), leaves (holy basil), stems (lemongrass) and rhizomes (turmeric) were separated from other parts and shade-dried in laboratory conditions (27±2°C, 60% RH) for seven days (Tapondjou et al., 2002). Dried herbs were ground into powder using an electric blender and sieved using a 0.84 mm (20 mesh) sieve by sieve shaker for several minutes. The herbs were put into a tea bag (5×4 cm) in 1 and 3 g/container, separately.
 
Volatile organic compounds (VOCs) analysis
 
Volatile organic compounds analysis on four natural fumigants were done at the Department of Biological Science and Technology, National Pingtung University of Science and Technology, Taiwan. This analysis was aimed to identify and determine of relative amounts of the VOCs from each fumigants. Samples were analyzed with solid-phase microextraction and gas chromatography-mass spectrometry (SPME-GC/MS) method as described by Cozzolino et al., (2016).
 
Insect biology test
 
The experiment was conducted during August­September 2018 at Integrated Pest Management and Urban Entomology Laboratory, National Pingtung University of Science and Technology, Taiwan. No choice test method was followed to assess the bioefficacy of the four natural fumigants on adult longevity, fecundity, F1 progeny, bean damage and weight loss of bean under completely randomized design (CRD) consisted of four fumigants (treated), positive control (without fumigant) and negative control (without fumigant and insect infestation) by replicating six times. Plastic container (Ø=6.5 cm, t=5 cm, v=±140 mL) filled with 30 g of adzuki bean was transferred with 15 pairs (0-24 hours old) of adults (Adedire and Akinneye, 2004; Righi-Assia et al., 2010; Regmi et al., 2012; Astuti et al., 2018) sexing the adult by elytral pattern (Beck and Blumer, 2014). The lid of plastic container was hermetically sealed with parafilm to prevent volatile escaping from the plastic container and it placed in laboratory conditions (27±2°C, 60% RH).

Adult mortality was assessed daily by counting the dead adults which paved way to arrive adult longevity. Insects were considered as dead if appendages (i.e. legs and antennae) did not move when touched with a paint brush for a minute (Hernandes-Lambrañoet_al2015). All dead adults were removed from the container.
       
Fecundity were counted by the eggs attached on surface of all beans after death of all released adults. Fecundity reduction was calculated and compared to control using the following formula (Gusmão et al., 2013) :
 
 

Whereas
Nc = Number of eggs (fecundity) on control.
Nt = Number of eggs (fecundity) on treatment.
 
F1 progeny were examined daily and dead adults if any inside of the beans were removed manually. F1 progeny reduction was calculated and compared with control using the following formula (Kedia et al., 2015) :
 

Whereas
Nc = Number of F1 progeny on control.
Nt = Number of F1 progeny on treatment.
 
 
Bean damage was calculated by observing emergence holes on the surface of beans using the following formula (Kedia et al., 2015) :
 
 
Whereas
Tb    = Total number of beans (damaged and undamaged).
Nud = Number of beans undamaged (bean with single or more holes counted as a bean damage).
 
       
Weight loss was calculated by the difference of weight before experiment started and the end of experiment and was calculated using the following formula (Kedia et al., 2015) :
 
 
Whereas
WI = Weight of beans at intial of experiment.
WE = Weight of beans at the end of experiment (negative control was used as correction factor to calculate at the end of each treatment).
 
Consumption reduction was observed by the difference of weight loss of beans on control and treatment and was calculated using the following formula (Kedia et al., 2015) :
 
 
Whereas
Wc = Weight loss of beans on control.
Wt = Weight loss of beans on treatment.

RESULTS AND DISCUSSION

Volatile Organic Compounds (VOCs) Analysis
 
Of the seven VOCs identified in clove powder, the major compounds were eugenol (48.64%) and caryophyllene (43.09 %) while 20 VOCs in holy basil powder with major compound as estragole (87.13%); 27 VOCs in lemongrass with a-Citral (28.88%), b-Citral (24.20%), (Z)-b-Caryophyllene (8.52%), selina-6-en-4-ol (6.85%) and estragole (5.51%) as major ones and 18 VOCs in turmeric powder with a-Zingiberene (35.22%), b-Sesquiphellandrene (20.18%), a-Curcumene (7.75%), eucalyptol (7.19%) and a-Turmerone (6.93%) as major compounds (Table 1).
 

Table 1: Volatile compounds of clove, holy basil, lemongrass and turmeric powder detected by SPME-GC/MS analysis.


 
Insect Biology Test
 
Clove (1 and 3 g dose) powder resulted in shorter adult  longevity of both sexes compared to other treatments (Table 2) besides reduced fecundity, number of F1 progeny, bean damage and weight loss of bean due to  clove (1 and 3 g dose) and holy basil (3 g dose) (Table 3 and 4). No adults (F1 progeny) emerged in these treatments leading to healthy adzuki bean without hole. Consumption reduction ranged 83-89% in clove and holy basil highly reduced the consumption in the beans.
 

Table 2: Efficacy of plant powders on adult longevity of Callosobruchus maculatus.


 

Table 3: Effficacy of plant powders on fecundity and F1 progeny of Callosobruchus maculatus.


 

Table 4: Efficacy of plant powders on bean damage and weight loss of bean of Callosobruchus maculatus.


 
The major volatile compounds that detected was a possible reason that plant powder affect the biological parameters of this species. In study on the toxicity of clove powder to red imported fire ants showed 100% mortality within 6 h (Kafle and Shih, 2013). Aslam et al., (2002) also confirmed that clove powder admixed with chickpea exhibited the shortest time to kill 100% of C. chinensis and significantly reducing the number of adult emergence and weight loss. Kafle and Shih (2013) confirmed the volatile compounds (eugenol, b-caryophyllene and eugenol acetate) has high toxicity on imported fire ants. In addition, eugenol has strong contact toxicity, reduce respiration rate of treated insects by fumigant and behavior modification on walking activity of granary weevil (Plata-Rueda et al., 2018). Other literature stated that each of major compounds that  contained in holy basil, lemongrass and turmeric powders were responsible for inhibition of oviposition, F1 progeny and mortality of many stored product pests (Bekele et al., 1996; Kim and Lee, 2014; Bossou et al., 2015).
 
Short adult longevity on clove, holy basil and lemongrass indicates that the insecticidal mode of action of the plant powder might be attributed to fumigant toxicity that penetrated the insect’s body through the respiratory system (spiracles and tracheas) (Adedire et al., 2011; Hamza et al., 2016). The inability of adults to mate and oviposit their eggs within surviving adult period that lead to the reduction of fecundity and F1 progeny numbers. Adedire et al., (2011) mention the botanical activities caused the disruption of locomotory insect that affected mating and ovipositing activities. Deterrent effects on F1 progeny that emerged into adults affected the postembryonic development that performed in less or even completely no adult emergence. This might have been caused by the VOCs from plant powders which poses the ability to penetrate the eggs chorion of insect through a minute space (micropyle) that is present on the posterior pole of eggs and results in the death of embryos and/or neonate larvae (Ileke et al., 2014; Kedia et al., 2015; Zafar et al., 2018). At the end of the experiment, little or no bean damaged and low weight loss of bean at certain plant powders were observed due to lack of insect survival stages such as insect infestation at the beginning of the experiment, eggs and neonate larvae against VOCs.

CONCLUSION

Holy basil, turmeric and lemongrass were effective against C. maculatus at 3 g dose. Among all treatments, though clove powder among the four treatments at 3 g and 1 g per container were most toxic by affecting the biological parameters of test insect. Four plant powders used in this study are good candidates to use in small scale storage facility to control the one of common species insect pests that attacked beans. Even though only two plant powders (clove and holy basil) at 3 g dose inhibited population growth, while other two plant powders (lemongrass and turmeric) only at 3 g dose, could inhibit the population growth. Of all these results, the synergistic action of VOCs released from each plant powder has contributed to the insecticidal properties against C. maculatus. These plant powders could be a good candidate to control stored-product pests in common traditional method in rural areas or small scale storage facility beside their role as grain protectants by contact toxicity.

ACKNOWLEDGEMENT

Authors would like to thank Double Degree Program between University of Brawijaya and National Pingtung University of Science and Technology. Also to Prof. Jeng-Tze Yang for suggestions in this research and Ito Fernando, M.Sc. for laboratory assistance.

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