m

ISBN No. 978-1-63315-205-2 (USA)

First Edition: Volume 1, 2 October 2014

Total No. Pages: 398 + 372 = 770

Edited by Raman Chandrasekar

B.K. Tyagi
Zhong Zheng Gui
Gerald R. Reeck
Copyright Reserved
Published by International Book Mission, Academic Publisher, South India.

Printed in the K-State Union, Copy and Printing services,

Kansas State University, Manhattan 66506, KS, USA.
This publication is considered to provide accurate and authoritative information with regards to the
subject matter has been obtained by its authors. The publisher has taken reasonable care in the
preparation of this book volume. However, the publisher and its authors shall in no event be liable for
any errors or omission arising out of use of this information and specifically disclaim any implied
warranties or merchantability or fitness for any particular use. No part of these books may be
reproduced, stored in retrieval system or transmitted in any form or by any means, electronic,
mechanical, photocopying or otherwide, without the prior permission of the Copyright owner.
Application for such permission, with a statement of the purpose and extend for the reproduction,
should be addressed to the publisher (IBM, Academic Publisher).
Price: US $ 250
Distributors and Subscription:

International Book Mission

Academic Publisher,
Nachiyar Silk and Printing House
76 Circuit House Road, 10th Cross NMK Colony
Tiruchirappalli 620 020, Tamilnadu,
South India.
Email: ibm_secretary@yahoo.com
ibmpublisher@gmail.com
ibmpublisher@yahoo.com
Tel. +91-431-2311187
Tel. +1-859-608-7694 (USA)

Book Mission Project # 2: Initiated on June 2010; Completed on March 2014 and Published on Oct. 2014.

Volume 1 & 2, October 2014

Short Views on Insect Biochemistry

and Molecular Biology

PREFACE
Entomology as a science of inter-depended branches like biochemistry, molecular entomology, insect
biotechnology; has made rapid progress in its attributes in the light of modern discoveries. This also
implies that there is an urgent need to manage the available resources scientifically for the good of man.
In the past five decades, entomology in the world/country has taken giant steps ahead. Continued
research has evolved better pest management through molecular approaches. The aim of the Short
Views on Insect Biochemistry and Molecular Biology book is to integrate perspectives across
biochemistry and molecular biology, physiology, immunology, molecular evolution, genetics,
developmental biology and reproduction of insects. This century is proclaimed as the Era of
Biotechnology and its consists of all types of Mol-Bio applications, which is an essential component for
a through understanding of the Insect Biology. This volume 1 & 2 (8 section with 30 chapters)
establishes a thorough understanding of physiological and biochemical functions of proteins, genes in
insects life processes; the topics dealt with in the individual chapters include chemistry of the insect
cuticle, hormone and growth regulators; biochemical defenses of insects; the biochemistry of the toxic
and detoxification action; modern molecular genetics and evolution; inter- and intra-specific chemical
communication and behavior; insect pheromone and molecular architecture, phylogeny and chemical
control of insect by using insect pheromones biotechnology; insect modern biology and novel plant
chemical and microbial insecticides for insect control, followed by a discussion of the various
mechanisms of resistance (both behavioral and physiological) and resistance management; modern insect
pest management through biochemical and molecular approaches; Mimetic analogs of insect
neuropeptide for pest management; entomo-informatics and computer-aided pesticide designing. In short
this book provides comprehensive reviews of recent research from various geographic areas around the
world and contributing authors area recognized experts (leading entomologist/scientist) in their
respective filed of molecular entomology. We will miss this collaboration now it has ended, but will feel
rewarded if this book is appreciated by our team/colleagues and remarkable mile stone in entomology
field.
This book emphasizes upon the need for and relevance of studying molecular aspects of entomology in
Universities, Agricultural Universities and other centers of molecular research. To encompass this
knowledge and, particularly disseminate it to the scientific community free of cost, was the major
inspiring force behind the launch of Short Views on Insect Biochemistry and Molecular Biology.

Editors

Raman Chandrasekar
Brij Kishore Tyagi

ii

iii

iv

vi

ShortViewson

InsectBiochemistryand
MolecularBiology
Editedby
Raman Chandrasekar, Ph.D.,
Kansas State University, USA.
B.K.Tyagi, Ph.D.,
Centre for Research in Medical Entomology (ICMR), India.
Zhong Zheng Gui, Ph.D.,
Jiangsu University of Science and Technology,
Sericultural Research Institute, Chinese Academy of
Agricultural Sciences, China.
Gerald R. Reeck, Ph.D.,
Kansas State University, USA.

vii

viii

SION
MIS

TERNA
IN
T

N AL B OO

IO

Contributing Authors

Dr. B.K.Tyagi

Prof.Fernando G. Noriega

Centre for Research in Medical Entomology,

4Sarojini Street, Chinna Chokkikulam,
Madurai 625002 (TN), India

Department of Biological Sciences

HLS 227, Florida International University
11200 SW 8th St, Miami, FL 33199, USA.

Prof. Gui Zhongzheng

Dr. Zhentao Sheng

Sericultural Research Institute,

Chinese Academy of Agricultural
Sciences, Zhenjiang, 212018,
Jiangsu, P. R. China.

Chicogo University, Chicogo, USA.

Prof. K. Sahayaraj

Prof.Yanyuan Bao
Institute of Insect Science,
Zhejiang University, China.

Dept. of Advanced Zoology and Biotechnology,

St. Xavier's College
Palayamkottai 627 002, Tamil Nadu, India.

Prof. Chuan-Xi Zhang,

Prof. David Siaussat

Dr. Maria L. Simes

Universit Pierre et Marie Curie (Paris 6/UPMC),

UMR 1272A Physiologie de l'Insecte:
Signalisation et Communication (PISC),
7 Quai Saint Bernard, Batiment A - 4me tage bureau 410, 75252 Paris Cedex 05, France.

Prof. PatriciaY.Scaraffia
Department of Tropical Medicine,
Tulane University, New Orleans,
LA 70112, USA.

Prof. Shou-An Xie

Institute of Insect Science,

Zhejiang University, China.

UEI Parasitologia Mdica,

Centro de Malria e Outras Doenas Tropicais,
Instituto de Higiene e Medicina Tropical,
Rua da Junqueira 96, 1300 Lisboa,
Portugal.

Dr. P. Somasundaram
Central Sericultural Germplasm Resources Centre,
P.B.No.44, Thally Road,
Hosur-635109,
Tamilnadu, India.

College of Forestry,
Northwest A & F University
Yangling, Shaanxi 712100, China

Dr. Hiroaki Abe

Dr. Raman Chandrasekar

Dr. S.K. Jalali

Department of Biochemistry and Molecular

Biophysics, Kanas State University,
Manhattan, 66506, KS, USA.

Prof. Gerald R. Reeck

Department of Biochem. and Molecular
Biophyscis, Kansas State University, KS, USA.

Prof. Manickam Sugumaran

Department of Biology
University of Massachusetts Boston
100 Morrissey Blvd,
Boston, MA 02125, USA.

Tokyo University of Agriculture and Technology,

Japan.

National Bureau of Agriculturally Important

Insects, ICAR, India.

Prof. Paraskeva V. Michailova

Institute of Biodiversity and
Ecosystem Research,
1 Tzar Osvoboditel boulv
Bulgarian Academy of Sciences
Sofia 1000, Bulgaria.

Prof. Ada Rafaeli

Associate Director for Academic Affairs &
International Cooperation
Agricultural Research Organization,
The Volcani Center, P. O. Box 6,
Bet Dagan 50250, Iseral.

ix

Prof. Emmanuelle Jacquin-Joly

Dr. Fei Liu

UMR PISC Physiologie de l'insecte

INRA, Route de Saint-Cyr
78026 Versailles cedex, France..

Department of Biological Science & Technol.,

Shaanxi Xueqian Normal University,
Shaanxi, China.

Dr. R. Srinivasan

Prof. Marian Goldsmith

Entomologist and Head of Entomology Group

AVRDC-The World Vegetable Center
60 Yi Ming Liao, Shanhua
Tainan 74151, Taiwan.

Biological Sciences Department,

University of Rhode Island,
Kingston, RI 02881, USA

Prof. Atanu Bhattacharyya

Prof. Anthony Ejiofor

Vidyasagar College for Women,

Post Graduate Department of Environmental
Science,
University of Kolkata, India.

Department of Biological Sciences,

College of Agriculture, Human & Natural
Sciences, Tennessee State University,
3500 John A Merritt Blvd., Nashville,
Tennessee 37209, USA.

Prof. Daphne Q.-D. Pham

Dr. Bharath Bhusan Patnaik

Dept of Biological Sciences,

University of Wisconsin-Parkside,
900 Wood Road, Kensoha,
WI 53144, USA.

School of Biotechnology,
Trident Academy of Creative Technology
(TACT), Bhubaneswar 751013 Odisha, India.

Prof. Jitrayut Jitonnom

Prof. B.R. Pittendrigh

School of Science
University of Phayao, Thailand.

Department of Entomology,
University of Illinois, Urbana-Champaign, IL,
61801, USA
.

Prof. K. Murugan

Dr. Subbiah Poopathi

Department of Zoology, School of Life Sciences,

Bharathiar University,
Coimbatore - 641 046, India.

Prof. Immo A. Hansen

Department of Biology,
New Mexico State University,
Las Cruces, NM, USA.

Dr. Ronald J. Nachman

USDA-ARS,
Food Animal Protection Research Laboratory,
USA.

Dr. Hari C Sharma

International Crops Research Institute for the
Semi-Arid Tropics (ICRISAT), Patancheru502324,
Andhra Pradesh, India.

Prof. Paolo Pelsoi

State Key Laboratory for Biology Plant Diseases
and Insect Pests, Institute of Plant Protection,
Chinease Academy of Agricultural Sciences,
Bejing, China.

Unit of Microbiology and Immunology,

Vector Control Research Centre
(Indian Council of Medical Research),
Medical complex, Indira Nagar,
Puducherry 60 5006, India.

Dr. P.Usha Rani

Biology and Biotechnology Division
Indian Institute of Chemical Technology
(CSIR)Taranaka,
Hyderabad - 500 007 (AP), India.

Dr. Fang Zhu

Irrigated Agriculture Research and Extension
Center, Dept.of Entomology,
Washington State University,
Prosser, WA, USA.

Prof. S.K.M. Habeeb

Department of Bioinformatics,
Faculty of Engineering & Technology,
SRM University, Kattankulathur,
Chennai 603203, Tamilnadu, India.

Prof. Yeon Soo Han

Division of Plant Biotechnology,
College of Agriculture & Life Science,
Chonnam National University,
Gwangju 500-757, South Korea

SION
MIS

TERNA
IN
T

N AL B OO

IO

Reviewer & External supportive members

Prof. Michael Riehle, Department of Entomology, University of Arizona, USA.

Dr. Dawn L.Geiser, College of Agriculture and Life Sciences, University of Arizona, USA.
Prof. Young Jung Kwon, School of Applied Biosci., Kyungpook National University, South Korea.
Dr. Kaliappandar Nellaiappan, CuriRx Inc. USA.
Prof. Patricia Y. Scaraffia, Department of Tropical Medicine, Tulane University, USA.
Prof. Richard Newcomb, Plant & Food Research, University of Auckland, New Zealand.
Dr. S. Krishnaswamy, School of Biotechnology, Madurai Kamaraj University, South India.
Dr. Mary-Anne Hartley, University of Lausanne, Switzerland.
Dr. Igor F. Zhimulev, Institute of Molecular and Cellular Biology, Novosibirsk, Russia.
Dr. S. Subramanin, Indian Agricultural Research Institute. India.
Prof. Gustavo F. Martins, Departament de Biologia Geral, Universidade Federal de Vicosa, Brazil.
Prof. Helena Janols, Infektionsklinien, Skanes Universitetsisjukhus, Sweden.
Prof. Donald R.Barnard, USDA, Agricultural Research Service, CMAVE, USA.
Dr. Keith White, Faculty of Life Science, University of Manchester, UK.
Prof. Marten J.Edwards, Biology Department, Muhlenberg College, USA.
Prof. E. Warchalowska-Sliwa, Polish Academy of Sciences, Poland.
Dr. K. Balakrishnan, Department of Immunology, Madurai Kamaraj University, India.
Dr. J.Joe Hull, USAD-ARS, Arid Land Agricultural Research Centre, USA.
Dr. Neil Audsley, The Food & Environment Research Agency, UK.
Dr. Raman Chandrasekar, Kansas State University, USA.
Dr. B.K. Tyagi, Centre for Research in Medical Entomology (ICMR), Madurai, TN, India.
Prof. Zhongzheng Gui, Sericulture Research Institute, Chinese Academy of Agricultural Sci., China.
Dr. Fang Zhu, Irrigated Agril. Research and Extension Center, Washington State University, USA.
Prof. K. Murugan, Department of Zoology, Bharathiar University, Coimbatore, India.
Dr. Xiao-Wei Wang, Institute of Insect Science, Zhejiang University, China.
Dr. Haijun Xu, Institute of Insect Science, Zhejiang University, China.
Dr. Alisha Anderson, CSIRO Ecosystem Sciences, Australia.
Prof. Eric D.Dodds, Department of Chemistry, University of Nebraska-Lincoln, USA.
Prof. P. Mosae Selvakumar, Department of Chemistry, Karnaya University, Coimbatore, India.
Prof. A.K.Dikshit, Indian Agriculture Research Institute, New Delhi.
Prof. K.R.S. Sambasiva Rao, Dept. of Biotech. & Zoology, Acharya Nagarjuna University, India
Dr. R. Rangeshwaran, National Bureau of Agriculturally Important Insects, Banglore, India.
Dr. V. Selvanarayanan, Faculty of Agriculture, Annamalai University, Tamil Nadu, India.
Prof. Fernando G. Noriega, Florida International University, Miami, USA.
Prof. Ada Rafaeli, Department of Food Quality and Safety, A.R.O., Israel.
Prof. Daphne Q.-D. Pham, Dept. of Biological Sciences, University of Wisconsin-Parkside, USA.
Prof. Emmanuelle Jacquin-Joly, INRA, UMR 1272 Physiologie de lInsecte, Versailles, France.
Prof. Manickam Sugumaran, University of Massachusetts Boston, USA.
Prof. Nannan Liu, Auburn University, USA.
Prof. Michihiro Kobyashi, Nagoya University, Japan.
Prof. Enoch Y.Park, Innovative Joint Research Center, Shizuoka University, Japan.
Prof. Luiz Paulo Moura ANDRIOLI, Universidade de So Paulo, SP - Brazil
Prof. SHIMADA Toru, The University of Tokyo, Japan.
Prof. Erjun Ling, Institute of Plant Physiology and Ecology, China.

xi

xii

Acknowledgements
Writing and publishing a book requires the assistance of individuals who are
creative, talented, and hard-working. All of these qualities were present in the
individuals assembled to produce this book volume. I would like to express my
heartfelt gratitude to my former teacher Prof. Seo Sook Jae, (GSNU, South Korea),
Prof. Subba Reddy Palli (University of Kentucky, USA), and other external mentors
Prof. Marian R. Goldsmith (University of Rhode Island, USA), Prof. Enoch Y. Park
(Shizuoka University, Japan), Prof. M. Kobayashi (Nagoya University, Japan), Prof.
CHU Jang Hann (National University of Singapore, Singapore), Prof. Thomas W.
Sappington (USDA-ARS, USA), Prof. Fernando G. Noriega (Florida International
University, USA), Dr. Srinivasan Ramasamy, AVRDC, The World Vegetable
Center, Taiwan), Dr. H.C. Sharam (ICRISAT, India), who inspiration and
supported me at many ways for the commencement of this International Book
Mission Program. The book mission program was initiated on May 2010,
completed on March 2014 and published on October 2014. I have no words to
express my feeling for all those who provided valuable contributions from USA,
South Korea, Japan, China, India, Thailand, Taiwan, Bulgaria, France, Iseral, and
Portugal (Contributors name list, see page no. v) and made the completion of this
book possible. We express our appreciation to the following people (Reviewer
name list, see page no. vii) who reviewed various part of the manuscript as it was
being developed and improved quality of each chapter. I thank the ICMR, New
Delhi, and Chinese Academy of Agricultural, China, and Kansas State University for
support from several aspects. Many others (scientists and publishers) have also
allowed us to use their materials in the various chapters, their color image have then
been converted to gray color/BW. Iam especially indebted to International Book
Mission Organization, Academic Publishing Services for the production of book. I
thank my Co-Editors for their continuous vigilance over the book project and for
always giving advance notice of the editing and proofreading schedules. I thank also
my Brintha, P.G., (my wife), who in all possible way, encouragement helped
transform our original efforts into an acceptable final form. I apologize to those
whose work could not be cited owing to space considerations limitation. Further, I
wish to recognize the moral support extended by colleagues and friends. I hope that
this volume will inspire interest on the diverse aspects of insect biochemistry and
molecular biology in aspiring and established scientists.
Raman Chandrasekar

xiii

xiv

A Note from the Publisher

Dear Readers,
This edition represents the first number of the Short Views on Insect
Biochemistry and Molecular Biology book series published by International
Book Mission. It serves to show the public how important entomology field in
expanding basic knowledge or in the development of new technologies nowadays,
in virtually all fields of knowledge. We called for piece of work falling into two
volumes (Basic and Advance aspects).
Far from being complete, the 30 chapters clearly structured and simply explained
experts contributions may provide an overview about current and prominent
advances in insect biochemistry and molecular biology which will help students and
researchers to broaden their knowledge and to gain an understanding of both the
challenges and the opportunities behind each approach.
We look forward to receiving new proposals for the new edition 2015 - 2017.
International Book Mission
Academic Publisher
Manager

xv

Book Series

xvi

Table Contents

SION
MIS

TERNA
IN
T

N AL B OO

IO

Page No.

Preface
Forward message
Contributors
Reviewers
Acknolwedgement

i
ii
iii
iv
v

Volume1
Section I: Insect Biochemical approaches

1. Introduction to Insect Molecular Biology.

Raman Chandrasekar, P.G., Brintha, Enoch Y.Park, Paolo Pelsoi, Fei Liu,
Marian Goldsmith, Anthony Ejiofor, B.R., Pittendrigh, Y.S., Han,
Fernando G. Noriega, Manickam Sugumaran, B.K., Tyagi, Zhong Zheng Gui,
Fang Zhu, Bharath Bhusan Patnaik, and P. Michailova

2.

Modulation of Botanicals on pests biochemistry.

57

Sahayaraj, K.

3.

Detoxication, stress and immune responses in insect antenna:

new insights from transcriptomics.

75

David Siaussat, Thomas Chertemps and Martine Maibeche

4.

Application of isotopically labeled compounds and tandem mass

spectrometry for studying metabolic pathways in mosquitoes.

99

Stacy Mazzalupo and PatriciaY.Scaraffia

5.

Field Response of Dendroctonus armandi Tsai & Li (Coleoptera:

Scolytinae) to Synthetic Semiochemicals in Shaanxi, China.

127

Shou-An Xie, Shu-Jie L.V., Hui-Chen, Raman Chandrasekar

xvii

Section II: Insect Growth

6. Insect Cuticular SclerotizationHardening Mechanisms and Enzymes.

149

Manickam Sugumaran

7. New Approaches to Study Juvenile Hormone Biosynthesis in Insects.

185

Crisalejandra Rivera-Perez, Marcela Nouzova and Fernando G. Noriega

8. The regulatory biosynthetic pathway of juvenile hormone.

217

Zhentao Sheng and Raman Chandrasekar

Section III:

Insect Immunity

9. The innate immune network in a hemimetabolous insect, the brown

planthopper, Nilaparvata lugens.

233

Yanyuan Bao, Raman Chandrasekar, Chuan-Xi Zhang

10. Immune Pathways in Anopheles gambiae.

253

Maria L. Simes and Raman Chandrasekar

11. Key biochemical markers in silkworms challenged with immuno-

271

elicitors and their association in genetic resistance for survival.

Somasundaram, P., Chandraskear, R., Kumar,K.A., and Manjula, A.

Section IV:

Insect Molecular Genetics

12. The recent progress of the W and Z chromosome studies of the

291

silkworm, Bombyx mori

Hiroaki Abe, Tsuguru Fujii and Raman Chandrasekar

13. Molecular characterization and DNA barcoding for identification of

317

agriculturally important insects.

Rakshit Ojha, Jalali, S.K., and Venkatesan, T.

14. Polytene chromosomes and their significance for Taxonomy,

331

Speciation and Genotoxicology

Paraskeva V. Michailova

15. Insect exuvium extracted DNA marker: a good complementary

molecular taxonomic characteristics with special reference
to mosquitoes.

355

Dhanenjeyan, K. J., Paramasivam, R., Thanmozhi, V., Chandrasekar,R., and Tyagi, B.K.
Index

363

xviii

Volume2
Section V:

Molecular Biology of Insect Pheromones

16. Understanding the functions of sex-peptide receptors?

373

Orly Hanin, Ada Rafaeli

17. Current views on the function and evolution of olfactory receptors

385

in Lepidoptera.

Arthur de Fouchier, Nicolas Montagn, Olivier Mirabeau, Emmanuelle Jacquin-Joly

18. Molecular architecture, phylogeny and biogeography of pheromone

409

biosynthesis and reception genes / proteins in Lepidoptera.

Jian-Cheng Chang, P. Malini, R. Srinivasan

Section VI:

Insect Molecular Biology

19. Application of Nanoparticles in sustainable Agriculture :

429

Its Current Status.

Atanu Bhattacharyya , Raman Chandrasekar, Asit Kumar Chandra,

Timothy T. Epidi and Prakasham, R.S.

20. Mosquito Ribonucleotide Reductase: A Site for Control.

449

Daphne Q.-D. Pham, Victor H. Perez, Lissette Velasquez, Dharty Bhakta,

Erica L. Berzin, Guoli Zhou, and Joy. J. Winzerling.

21. Green protocol for synthesis of metal nanoparticles

to control insect pests.

473

Murugan, K., Chandrasekar, R., Panneerselvam, C., Naresh Kumar, A.,

Madhiyazhagan, P., Mahesh Kumar, P., Jiang-Shiou Hwang, Jiang Wei

22. Aquaporins in Blood-Feeding Arthropods.

497

Lisa L. Drake, Hitoshi Tsujimoto, Immo A. Hansen

23. Mimetic analogs of three insect neuropeptide classes

509

for pest management.

Ronald J. Nachman

xix

Section VII:

Insect Pest Management through

Biochemical and Molecular approaches

24. Induced resistance in plants and counter-adaptation by insect pests.

533

Abdul Rashid War and Hari C Sharma

25. Insect Chemical communication - an important component of

549

novel approaches to insect pest management.

Usha Rani, P.

26. Mosquito control using biological larvicides: Current Scenario.

575

Subbiah Poopathi, C. Mani and R. Chandrasekar

27. Application of RNAi toward insecticide resistance management.

595

Fang Zhu, Yingjun Cui, Douglas B. Walsh, Laura C. Lavine

Section VIII:

Insect Bioinformatics

28. Entomo-informatics: A prelude to the concepts in Bioinformatics.

621

Habeeb, S.K.M. and Raman Chandrasekar

29. Molecular expression and structure-function relationships of

633

apolipophorin III in insects with special reference to innate immunity.

Bharat Bhusan Patnaik, Raman Chandrasekar, Yeon Soo Han

30. Computer-aided pesticide design: A short view

685

Jitrayut Jitonnom

Index

709

xx

ShortViewson
InsectBiochemistryand
MolecularBiology

Volume 1 & 2

ABSTRACTS

Volume1
Abstract

Chapter -1

Introduction to Insect Biochemistry

and Molecular Biology
Raman Chandrasekar1*, P.G., Brintha2, Enoch Y. Park3,
Paolo Pelsoi4, Fei Liu5, Marian Goldsmith6, Anthony
Ejiofor7, B.R., Pittendrigh8, Y.S., Han9, Fernando G.
Noriega10, Manickam Sugumaran11, B.K., Tyagi12,
Zhong Zheng Gui13, Fang Zhu14, Bharath Bhusan
Patnaik9,15, and P. Michailova16
1

Department of Biochemistry and Molecular Biophysics, 238 Burt Hall, Kansas State
University, Manhattan, KS, USA
2
Department of Biology, Kansas State University, Manhattan, KS, USA
3
Green Chemistry Research Division, Research Institute of Science and Technology,
ShizuokaUniversity, 836 Ohya Suruga-ku Shizuoka 422-8529, Japan
4
State Key Laboratory for Biology Plant Diseases and Insect Pests, Institute of Plant
Protection, Chinease Academy of Agricultural Sciences, Bejing,China
5
Department of Biological Science and Technology, Shaanxi Xueqian Normal
University, Xian, Shaanxi, China
6
Biological Sciences Department, University of Rhode Island, Kingston, RI 02881, USA
7
Department of Biological Sciences, College of Agriculture, Human & Natural
Sciences, Tennessee State University, 3500 John A Merritt Blvd., Nashville,
Tennessee 37209, USA
8
Department of Entomology, University of Illinois, Urbana-Champaign, IL, 61801, USA
9
Division of Plant Biotechnology, College of Agriculture & Life Science, Chonnam
National University, Gwangju 500-757, South Korea
10
Department of Biological Sciences, Florida International University, Miami,
FL 33199, USA
11
Department of Biology, University of Massachusetts Boston, Boston, MA 02125, USA
12
Centre for Research in Medical Entomology (ICMR), 4-Sarojini Street, Chinna
Chokkikulam, Madurai 625002, TN, India
13
Jiangsu University of Science and Technology, Sericultural Research Institute,
Chinese Academy of Agricultural Sciences, China
14
Department of Entomology, Washington State University, Pullman, WA, USA
15
School of Biotechnology, Trident Academy of Creative Technology (TACT),
Bhubaneswar 751013 Odisha, India
16
Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of
Sciences, Bulgaria

Entomology now is a diversified science discipline,

deviating considerably from the incorporated principles
of Molecular biology, Genetics and Biochemistry. It has
provided necessary tools for transferring and evaluating
genetic characteristics not only for a host of insects, but
also for related host plants. The molecular approaches
have enabled the study of physiologicaly vital
proteins/enzymes and sensillar-neural complexes that
are involved in pheromonal studies. The knowledge is
vital to devise safe and specific agents for disrupting
insect life cycles, thus increasing the efficiency of efforts
to manage agricultural pests and disease vectors. This
chapter analyzes the impact of new technologies that
allow study of molecular events at the single cell level,
and highlights the need of modern insect biological
research on insect systems.

Keywords: entomo-informatics, biotechnology, molecular

biology, genetics, RNA interference, genomics, proteomics,
pheromone
*For Correspondence (email: biochandrus@yahoo.com)

Chapter-2
Modulation of Botanicals on Pests
Biochemistry
K. Sahayaraj*
Department of Advanced Zoology and Biotechnology, St. Xavier's College
(Autonomous), Palayamkottai 627 002, Tamil Nadu, India
Abstract

Insect lives in any agro-ecosystem both as beneficial and

harmful organisms to crops. They represent within
insect orders particularly Lepidoptera, Hemiptera,
Orthoptera, Coleoptera, Thysanura, Blateria ect. The
main consequence of insect pests includes infestation as
well as acts as vectors. These problems were apparently
under control after the development of conventional
insecticides. However, side effects have been notices for
microbes, plants, animals, and our environments. One
among the well-known alternative is botanicals. Some
studies that have searched for potential anti-insect
products from plants have focused on repellent activity
to prevent insect pests infestation. Most studies,
however, evaluated the toxicity of plant extracts and
plant-derived compounds, in order to control insect
pests. To our knowledge, no attempts have been made
to discuss the biochemical impact of botanicals against
pests. In this review we focus on impacts of botanicals
on carbohydrate (glycogen and trehalose), proteins (total
proteins and amino acids), lipids (total cholesterols and
phospholipids), electrolytes (Na+, Cl, K+ and Ca2+
levels) and both digestive (Amylase, Glycosidases,
Alkaline phosphatase, Lipases, Proteases) and
detoxication (esterase, oxidase, transoxidase hydrolases,
glutothion transferase, cis-oxidase hydrolases, adrin
epoxidase, cytochrome P-450, alanine aminotransferase,
asparate amino- teransferase, glutathione S-transferase,
glutathione P-transferase) enzymes either directly or
indirectly to cause death or alter the behaviors which
lead to death.
Key words: biochemical analysis, carbohydrates, lipid,
enzymes, detoxification, mortality, plant defense
*For Correspondence (email: ksraj42@gmail.com)

Chapter -3
Detoxication, stress and immune
responses in insect antenna: new
insights from transcriptomics
David Siaussat*, Thomas Chertemps, Martine
Mabche-Coisne
UMR 1272 UPMC-INRA PISC Physiologie de lInsecte: Signalisation et
Communication,7 Quai Saint Bernard, F-75000 Paris, France.
Abstract

Studies on the insect olfactory peripheral system are

mainly focused on deciphering the receptor and
perireceptor events at the molecular, cellular and
physiological levels. The other processes that could
occur in the antennae were much less studied. However,
as antennae are directly in contact with the outer
environment, with no physical barrier protecting the
sensory neurons except the sensillar lymph, one can
hypothesize the existence of antennal mechanisms of
adaptation to a variety of environmental stresses. Recent
transcriptomic and proteomic approaches in various
species highlighted the expression in antennae of many
genes putatively involved in stress response. Using data

available from several antennal transcriptomes, we

identified a diversity of genes already known for their
involvement in detoxication, the stress and immune
responses in other tissues and we discussed their
possible roles within the olfactory organ. Our analysis
strongly suggests that insect antenna has evolved various
mechanisms of defence, as already known for vertebrate
olfactory mucosa.
Key words: Insect, antennae, transcriptome, proteome,
stress response, immunity, enzymes
*For Correspondence (email: david.siaussat@upmc.fr)

traditional and cutting-edge approaches applied to the

study of nitrogen metabolism in mosquitoes are
presented in this chapter. Schemes of recently proposed
metabolic pathways are also shown. The discovery of
ammonia fixation, assimilation and excretion pathways
occurring in tandem illustrates the complexity of
mosquito biology, yet also provides evidence that a
better understanding of the metabolic processes that
operate in mosquitoes is still necessary for the discovery
of new targets that will lead to successful control of
mosquito populations.
Key words: Metabolites, selective reaction monitoring,
sample
preparation,
insects,
labeled
compounds,
derivatization, nitrogen waste, gene expression, RNA
interference
*For Correspondence (email: pscaraff@tulane.edu)

Chapter-4
Application of isotopically labeled
compounds and tandem mass
spectrometry for studying metabolic
pathways in mosquitoes
Stacy Mazzalupo1 and Patricia Y. Scaraffia1,2*
1

Department of Chemistry and Biochemistry, and The Center for Insect

Science, The University of Arizona, Tucson, AZ 85721,USA
2
Current address: Department of Tropical Medicine, Tulane University,
New Orleans, LA 70112, USA
Abstract

More than one million deaths occur annually due to

mosquito-borne diseases. To resolve this devastating
worldwide problem many researchers are trying to find
new metabolic targets for vector control. In this chapter,
we focus upon one mosquito species, Aedes aegypti, and
discuss how classic and modern biochemistry, such as
the application of isotopically labeled compounds and
mass spectrometry methods, have impacted our
understanding of mosquito biology. Recent studies
demonstrate that A. aegypti female mosquitoes have very
efficient mechanisms to metabolize the amino acids and
ammonia produced during the digestion of a blood meal.
An overview of the metabolism of blood meal protein
amino acids and a comprehensive discussion of

Chapter-5
Field Response of Dendroctonus
armandi Tsai & Li (Coleoptera:
Scolytinae) to Synthetic
Semiochemicals in Shaanxi, China
Shou-An Xie1*, Shu-Jie LV1, Hui-Chen1, Raman
Chandrasekar2
1

College of Forestry, Northwest A & F University, Yangling, Shaanxi

712100, China
2
Department of Biochemistry & Molecular Biophysics, Kansas State
University, Manhattan 66506, KS, USA

Abstract

Bark beetles (Coleoptera: Curculionidae: Scolytinae) are

economically and ecologically the most important group
of forest insects. The pine bark beetle, Dendrotonus
armandi is native to Qinling and Bashan Mountains in
China. Its host tree is Pinus armandii, which occurs in

previous two mountain ranges and other mountain areas

of Southwestern China. This beetle is a unique species,
different from all others in the genus. It mainly attacks
apparently healthy trees over age 30, and has recently
caused serious damage to the P. armandii stands in the
Qinling Mountains. We hypothesized that there are
differences between the numbers of beetles captured
with traps baited with various combinations of
candidate semiochemicals extracted from the hindguts
of D. armandi. In order to determine whether a better
operational lure could be developed for D. armandi, we
analyzed the volatiles in extracts of the hindgut of D.
armandi and tested various combinations of candidate
semiochemicals. The GC-MS and GC-FID analyses of
volatiles in the extracts of the hindguts of D. armandi
collected at different stages of the attack revealed that
they are mainly -pinene, -caryophyllene and (+)-3carene with minor amounts of myrcene, limonene,
verbenol and verbenone. We tested various
combinations of these candidate semiochemicals in
order to determine an optimal blend. Our results suggest
that the addition of -caryophyllene to either -pinene,
or blends of -pinene and other candidate
semiochemicals, significantly enhanced the attractiveness
of the lures for D. armandi. Field trapping experiments
indicated that the blends that included -caryophyllene,
myrcene and other candidate semiochemicals resulted in
significantly higher trap catches than -pinene alone.
Therefore, a simple lure consisting of -pinene and caryophyllene would be an optimal blend for D. armandi.
We conclude that this blend of semiochemicals may
provide a better method of reducing the numbers of D.
armandi in the forest ecosystem.
Key words: Dendroctonus armandi; semiochemicals; GCMS and GC-FID; trap; Pinus armandi; Biochemistry
*For Correspondence (email: shouanxie@163.com;
shouanxie@126.com )

Chapter-6
Insect Cuticular Sclerotization
Hardening Mechanisms and
Enzymes
Manickam Sugumaran*
Department of Biology, University of Massachusetts Boston, 100 Morrissey
Blvd, Boston, MA 02125, USA
Abstract

Sclerotization of insect cuticle is an extremely important

and vital biochemical process for the survival of all
insects. In this review, the biochemistry, enzymology
and molecular mechanism of cuticular sclerotization are
critically examined. A unified sclerotization mechanism
involving the participation of quinone, quinone methide,
quinone methide imine amide, and semiquinones as
reactive intermediates of sclerotization is presented. The
key role played by cuticular enzymes such as odiphenoloxidases, laccases, quinone isomerase, quinone
methide isomerase and peroxidase in sclerotization
process is discussed. Cuticular sclerotization process can
provide a unique tool to develop new kinds of insect
control measures that are highly specific for target
insects.
Key words: cuticular sclerotization, quinone tanning,
quinone methide sclerotization, quinone methide imine
amide, free radical tanning, phenoloxidases, laccases,
tyrosinases, quinone isomerase, quinone methide isomerase
*For Correspondence
(email: manickam.sugumaran@umb.edu)

Chapter-8

Chapter-7
New Approaches to Study Juvenile
Hormone Biosynthesis in Insects
Crisalejandra Rivera-Perez, Marcela Nouzova, and
Fernando G. Noriega*
Department of Biological Sciences, Florida International University,
Miami, FL 33199,USA
Abstract

The juvenile hormones (JHs) represent a family of

sesquiterpenoids that play a central role in insect
reproduction, development and behavior. The lipophilic
character of JHs, their low concentration in tissues and
fragility had made their measurement difficult. Four
main methods have been used for JH quantification:
bioassays, radioimmunoassays, radiochemical assays and
physicochemical assays. Among the physicochemical
assays, mass spectroscopy (MS) and high performance
liquid chromatography coupled to a fluorescent detector
(HPLC-FD) represent the best techniques for accurate
identification and sensitive quantification of JHs in
biological samples. This chapter aims to provide an
overview of JH biosynthesis in insects, with an emphasis
on two main aspects: 1) the techniques used to study
juvenile hormone synthesis and 2) the development of
new approaches for the quantification of JH precursors.
Key words: Juvenile hormone, biosynthesis, precursors,
farnesoic acid, mosquitos, fluorescent tags, HPLC-FD.
*For Correspondence (email: noriegaf@fiu.edu )

The regulatory biosynthetic pathway

of juvenile hormone
Zhentao Sheng1*, Raman Chandrasekar2
1

The Ben May Department for Cancer research, The University of

Chicago, Chicago, IL, 60637, USA.
2
Department of Biochemistry & Molecular Biophysics, Kansas State
University, Manhattan, KS 66506, USA
Abstract

In this short review, it will be focused on the regulatory

biosynthetic pathway of juvenile hormone (JH). JH is
involved in the regulation of nearly every aspect of
insect life, such as metamorphosis, reproduction,
diapause, lifespan and polyphenisms. To perform these
biological functions, the biosynthesis of JH is under a
rigorous control by insulin/TOR signaling pathway,
neuropeptides, neurotransmitters and other bioactive
molecules, modulating enzymes in the JH biosynthetic
pathway. JH acid methyl transferase (JHAMT) converts
JH acid to JH or farnesoic acid (FA) to methyl
farnesoate (MF) at final steps in JH biosynthesis.
JHAMT is the key regulatory enzyme for JH
biosynthesis, which is activated by DPP-mediated TGF
signaling and also targeted by other regulators for JH
biosynthesis. Targeting genes in JH synthetic pathway,
especially the late insect specific steps, was already
broadly applied in studies on JH function and is a
promising strategy for pest control.
Key words: juvenile hormone, JH acid methyl transferase,
Neuropeptides, Insulin/TOR signaling pathway
*For Correspondence (email: zsheng@uchicago.edu or
zhtsheng911@gmail.com)

Chapter-9
The innate immune network in a
hemimetabolous insect, the brown
planthopper, Nilaparvata lugens
Yan-Yuan Bao1*, Raman Chandrasekar2, Chuan-Xi
Zhang1*
1

State Key Laboratory of Rice Biology and Ministry of Agriculture,

Key Laboratory of Agricultural Entomology, Institute of Insect Sciences,
Zhejiang University, Hangzhou 310058, China
2
Department of Biochemistry and Molecular Biophysics, 238, Burt Hall,
Kansas State University, Manhattan 66506, KS, USA
Abstract

Here we performed a genome-wide analysis aiming at

the immunity in a hemimetabolous insect, the brown
planthopper. We identified a large number of genes
encoding N. lugens pattern recognition proteins,
modulation proteins in the prophenoloxidase (proPO)
activating cascade, immune effectors and signal
transduction molecules involved in the immune
pathways, including the Toll, Immune deficiency (Imd),
Janus kinase signal transducers and activators of
transcription (JAK-STAT) and c-Jun N-terminal kinases
(JNK) pathways. A comparison of the genome-available
hemimetabolous and holometabolous insect species
indicate the differences in the immune-related gene
constitution. The genome-wide analysis of immunerelated genes is an important step in determining the
overall architecture and functional network of the
immune components in N. lugens, which could facilitate
the understanding of the innate immune mechanisms in
the hemimetabolous insect species.
Key words: Nilaparvata lugens, hemimetabolous insect,
genome; innate immunity, signal pathway, Antimicrobial
protein
*For Correspondence
(yyabo@zju.edu.cn and chxzhang@zju.edu.cn )

Chapter-10
Immune Pathways in Anopheles

gambiae

Maria Lusa Simes1*, and Raman Chandrasekar 2

1

UEI Parasitologia Mdica, Centro de Malria e Outras Doenas

Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de
Lisboa, Lisboa, Portugal
2
Department of Biochemistry and Molecular Biophysics, 238 Burt Hall,
Kansas State University, Manhattan 66502, KS, USA
Abstract

Several experimental studies have demonstrated that

Anopheles mosquitoes are able to mount an efficient
immune response against Plasmodium infection, and that
this results in major losses for the parasite during its
development inside the vector. The mosquito immune
defence involves complex mechanisms of action, which
have been intensively researched during the last decade,
and have led to the development of novel interventions
against malaria based on vector-parasite interactions. In
this review, we describe the main components of the
innate immune system in Anopheles gambiae and how they
impact Plasmodium development. We focus our attention
on the two major immune signalling pathways, Toll and
Imd, and their role in controlling the transcription and
expression of genes described as anti-Plasmodium
effectors for their activity in fighting the pathogen
responsible for one of the worlds deadliest diseases.
Key words: Anopheles, Plasmodium, malaria, innate
immunity, PAMPs, PRRs, Toll and Imd pathways, gene
expression, AMPs, transgenic mosquitoes.
*For Correspondence
(email: maria.luisa.simoes@ihmt.unl.pt)

Chapter-11
Key biochemical markers in
silkworms challenged with immuno
elicitors and their association in
genetic resistance for survival
Somasundaram, P.1*, Raman Chandrasekar2, Ashok
Kumar, K.1, Manjula, A.1
1

Central Sericultural Germplasm Resources Centre, P.B.No.44,

Thally Road, Hosur-635109, Tamilnadu, India
2
Department of Biochemistry and Molecular Biophysics, 238, Burt Hall,
Kansas State University, Manhattan KS 66506, USA
Abstract

Sericigenous insects like silkworm possess an effective

immune system against pathogens. The knowledge on
the immune system of these insects implicating the
mechanism of resistance to diseases as mainly
attributable to the presence of key biomolecules such as
an inducible proteins viz., cecropin, attacin, lebocin
,moricin and isoforms of enzymes viz., carboxyl esterase
and prophenoloxidase, which are reproducible
qualitative biochemical markers that would favor us in
enhancing our theoretical level on the mechanism of
immunity. Studies on these key biomolecules in
silkworms challenged with immuno elicitor viz.,
lipopolysaccharide (LPS) as a function of immunity and
survival ability of the silkworms revealed the mode of
cellular and humoral functions combating the diseases in
nature. These factors can be used by researchers or
breeders to identify the appropriate hardy genetic
material from the germplasm stock for their further
breeding work. In this review an attempt has been made
to delineate those key biomolelcules which are
qualitatively reproducible and that support survival of
the silkworms of Bombyx mori (L). These factors may be
considered as key factors to identify the genetic
resistance of various silkworm races/breeds of B. mori
and documentation of hardy silkworm breeds for field
exploitation in different climatic conditions of the
tropical zones of the country.
Key words: silkworm, resistance, inducible proteins,
biomolecules, germplasm, survival, prophenoloxidase,
Bombyx mori
*For Correspondence (email: drpsunder@gmail.com)

Chapter-12
The recent progress of the W and Z
chromosome of the silkworm,

Bombyx mori

Hiroaki Abe1*, Tsuguru Fujii2 and Raman

Chandrasekar3
1

Department of Biological Production, Faculty of Agriculture, Tokyo

University of Agriculture and Technology, Saiwai-cho 3-5-8,183-8509,
Fuchu, Tokyo, Japan
2
Institute of Genetic Resources, Graduate School of Bio Resources and
Bioenvironmental Science, Kyushu University, Higashi-ku,
Fukuoka 812-8581, Japan
3
Department of Biochemistry and Molecular Biophysics, 238 Burt Hall,
Kansas State University, Manhattan 66506, KS, USA
.
Abstract

Sex chromosomes of the silkworm, Bombyx mori, are

designated as ZW (XY) for females and ZZ (XX) for
males. The female mode of development is determined
by the presence of a single W chromosome. Therefore, it
is presumed that the female-determining gene (Fem) is
present on the W chromosome. Until date, 12 Wspecific random amplified polymorphic DNA (RAPD)
markers have been identified on the normal W
chromosome. We have compared the W-translocation
chromosomes with normal W chromosome. The
T(W;3)Ze (the sex-limited Zebra strain) chromosome
lacked 2 W-specific RAPD markers, indicating that the
region containing these two W-specific RAPD markers
has been deleted. Additionally, we have investigated
T(W;2)Y (the sex-limited Yellow-cocoon strain). The
T(W;2)Y chromosome lacked 11 such markers. These
results indicate that the W chromosomes of the sexlimited strains are the products of reciprocal

translocation accompanied by deletion, and an extremely

limited region is required to determine femaleness. No
genes for morphological traits have been mapped to the
W chromosome. Contrary, Z chromosome is rich in
genes. Many long terminal repeat (LTR) and non-LTR
retrotransposons, retroposons, DNA transposons, and
their derivatives, had accumulated as strata on the W
chromosome. It is notable that some of these
transposable elements contained the Bombyx short
interspersed element (Bm1) sequences in the elements.
On the other hand, the transposable elements on the Z
chromosome were excluded by unequal crossing over or
intra-element homologous recombination between
LTRs.
Key words: Bombyx mori, female-determining gene,
transposable elements, single nucleotide polymorphism,
random amplified polymorphic DNA, W chromosomes, Z
chromosomes
_________________________________________________
*For Correspondence (email: wfem@cc.tuat.ac.jp)

Abstract

Insects collected from various ecosystems in several

places in India were sequenced and barcoded for a
658bp region of the mitochondrial cytochrome c oxidase
subunit 1 gene (cox1). This gene is frequently termed as
CO1 in barcoding approach and serves as the core of a
global bio-identification gene for insects and other
animals. It is estimated that in India approximately
130000 insect species are available, of which about
60000 insect species have been identified. However, only
2008 species have been barcoded, which is 3.34% of
identified species, whereas at world level about 12.3%
species have been barcoded so far. To meet this
daunting challenge of quick and reliable insect
identification, DNA barcoding is now been considered
as an alternate tool for insect biodiversity identification
in India and the world. The present chapter deals with
the use of barcode in the identification of 42 insects
belonging to 5 orders and 15 families and using
Neighbor-joining approach with bootstrapping method
and Kimura-2 parameter to obtain a clear phylogenetic
signal. In a neighbor-joining tree for all sequences, two
clades were obtained, the first cluster consisting of
orders Lepidoptera, Diptera, Hemiptera and Coleoptera,
whereas another clade showing relationship between
Hymenopteran insects, which is not in contradiction
with cladistics analysis of morphological data for insect
orders and is consistent with traditional phylogeny of
insects. The present results thus favour DNA barcoding
as a decisive tool in quick and reliable identification of
Insects.
Key words: Insects, DNA barcoding, phylogeny, CO1,
India, mitochondrial markers, taxonomy
_________________________________________________
*For Correspondence (email: jalalisk1910@yahoo.co.in )

Chapter-13
Molecular characterization and DNA
barcoding for identification of
agriculturally important insects
Rakshit Ojha, S. K. Jalali* and T. Venkatesan
National Bureau of Agriculturally Important Insects, Post Bag No. 2491,
H. A. Farm Post, Hebbal, Bellary Road, Bangalore 560024, Karnataka,
India

Chapter-14
Polytene chromosomes and their
significance for Taxonomy,
Speciation and Genotoxicology
Paraskeva Michailova*
Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of
Sciences, 1, Blvd. Tsar Osvoboditel, 1000 Sofia, Bulgaria

Chapter-15

Abstract

The
polytene
chromosomes
are
interphase
chromosomes which are formed by the endomitotic
replication of chromosomes without cell division. Their
large size, constant somatic pairing of the homologues
and species-specific band patterns makes them
important markers for taxonomy, systematics and
evolution. The significance of the polytene
chromosomes for the taxonomy and systematics of the
family Chironomidae is discussed. Interspecies
differences by chromosome rearrangements, localization
and appearance of heterochromatin, differences in
amount of repetitive DNA elements are shown in the
genera Chironomus Mg. and Glyptotendipes Kieffer. The
extent and under what circumstances changes in the
banding patterns of the polytene chromosomes may
initiate sympatric, allopatric speciation and introgressive
hybridization are examined. The presence of fixed
aberrations in the homozygous state is an important
factor of species divergence. Sympatric species display
more complicated chromosome rearrangements then
allopatric species. In addition, hybridization tests
revealed that pre - and postmating isolating mechanisms
were operating. The introgressive hybridization in
chironomids, a process of incorporation of genes of one
species into the gene pool of another species was
shown. The high resolution of chironomid salivary
gland chromosomes and their sensitivity to external
stressors allows their use in the diagnosis of
environmental quality and hence of the potential
hazards posed by environmental pollution.
Key words: Polytene chromosomes, Chironomidae,
sympatric and allopatric speciation, introgressive
hybridization, genotoxicology
________________________________________________
*For Correspondence (email: michailova@zoology.bas.bg
or pmichailova@yahoo.com)

Insect exuvium extracted DNA

marker: a good complementary
molecular taxonomic characteristics
with special reference to mosquitoes
K.J. Dhananjeyan1, R. Paramasivan1, V.
Thanmozhi1, R.Chandrasekar2 and B.K. Tyagi1*
1

Centre for Research in Medical Entomology (ICMR), 4-Sarojini

Street, Chinna Chokkikulam, Madurai 625002, TN, India
2
Department of Biochemistry and Molecular Biophysics,
238 Burt Hall, Kansas State Univeristy, Manhattan, KS, USA
Abstract

Many a insect species, particularly sibling species

complexes and those sylvatic taxa whose adults are
difficult to catch with information on breeding habitats,
pose challenge to their correct identification. Many
genetical and molecular techniques have earlier been
attempted on mosquito species, but without much
success in complimenting existing dichotomous keys.
Taking cues from a major molecular taxonomic
breakthrough in odonata, we have adopted this
technique refered to as application of genomic DNA
extracted from nondestructible structures in mosquito
taxonomy of important Culex, Anopheles and Aedes
species.
Key words: Taxonomy, DNA marker, mosquitoes,
exuvium, cytochrome oxidase,
_________________________________________________
*For Correspondence (email: bk_tyagi@sify.com)

Volume2

Chapter-16
Understanding the functions of sex
peptide receptors?
Orly Hanin1,2 and Ada Rafaeli2*
1

Department of Entomology, Hebrew University, Rehovot, Israel

Department of Food Quality and Safety, Volcani Center,
Agricultural Research Organization, Bet Dagan, Israel

Abstract

In many insect species reproductive success is largely

dependent on synchronized behavioral and physiological
activities of both sexes. Mating behavior is often
perceived as an arms race between the sexes; whereas
the males' interests lie in being the sole mate, the
females' interests oppose this monogamous activity in
the search for the "successful" partner to sire her
offspring. Within this framework of evolutionary
selection pressures, males of most insects produce and
transfer peptides during copulation that affect the
females' reproductive behavior to induce a reduction in
receptivity to other mating attempts and an increase in
the rate of oviposition. An example of such a transferred
peptide is the Drosophila melanogaster sex-peptide
responsible for post-mating changes in the female. In
this review, we present the current findings concerning
the functions of sex-peptide and sex-peptide receptors
and provide possible insights as to the evolutionary
constraints that may exist between the ligand and its
receptor.
Key words: sexual behavior, receptivity, oviposition, sexpeptide, myoinhibitory peptides, Helicoverpa armigera,
Drosophila melanogaster
_________________________________________________
*For Correspondence (email: vtada@volcani.agri.gov.il )

Chapter-17
Current views on the function and
evolution of olfactory receptors in
Lepidoptera
Arthur de Fouchier1, Nicolas Montagn2, Olivier
Mirabeau1 and Emmanuelle Jacquin-Joly1,*
1

INRA, UMR 1272 Physiologie de lInsecte : Signalisation et

Communication, Versailles, France
2
UPMC-Universit Paris 6, UMR 1272 Physiologie de lInsecte :
Signalisation et Communication, Paris, France
Abstract

The sense of olfaction stimulates many vital behaviors in

insects. At the molecular level, the interactions between
an insect and its olfactory environment are mediated by
two families of chemosensory receptors, the olfactory
receptors and the ionotropic receptors. In this chapter,
we review the current knowledge on olfactory receptors
within the Lepidoptera. We expose the different
strategies used for their identification, via genome and
transcriptome sequencing, and describe the principles
underpinning the different in vitro and in vivo assays

developed for their functional characterization. While

hundreds of sequences have been annotated as olfactory
receptors, only a small number have been deorphanized
and most of these are pheromone receptors. So far the
data suggest a combinatorial model of odor coding as
revealed in Drosophila. From an evolutionary point of
view, several highly conserved clades of olfactory
receptors can be defined within Lepidoptera,
representing ancestral paralogous lineages, with
functional divergence observed within some lineages. In
the near future, we expect the characterization of large
repertoires of Lepidoptera olfactory receptors, which
will shed new light on how evolution has tuned olfaction
in different species according to their individual needs
and niches.
Key words: Olfactory receptor, odorant-binding proteins,
Lepidoptera, molecular evolution
_________________________________________________
*For Correspondence
(email: emmanuelle.jacquin@versailles.inra.fr)

Chapter-18
Molecular architecture, phylogeny
and biogeography of pheromone
biosynthesis and reception genes /
proteins in Lepidoptera
Jian-Cheng Chang, P. Malini and R. Srinivasan*
AVRDC The World Vegetable Center, P. O. Box 42, Shanhua, 74199
Tainan, Taiwan
Abstract

The stimulation, regulation, and detection of

semiochemicals including sex pheromones is essential
for insects to select hosts, find food, evade natural
enemies and harmful materials in the environment, and
seek con-specific mating targets. Female moths, for
example, release sex pheromones to attract appropriate
males for mating in the field. While the biosynthesis of
female sex pheromone is induced by a neuropeptide
hormone,
pheromone
biosynthesis
activating
neuropeptide (PBAN), the water-soluble pheromone
binding proteins (PBPs) play a critical role in
transporting sex pheromone components through
sensillar lymph to olfactory receptor neurons in males.
Lepidopteran PBAN is a polyprotein comprising five
putative neuropeptides: diapause hormone (DH),
PBAN, -subesophageal ganglion neuropeptide (SGNP),
-SGNP and -SGNP. These peptides share a common
FXPRLamide in the C-termini. PBAN neuropeptides are
well conserved across lepidopteran species and extend to
the Insecta. PBPs are proteins of 120150 amino acids.
Multiple-PBPs are known to occur in the same moth
species. Although they display considerable diversity,
they are related in sequence. They play a vital role in
discrimination of pheromone components. In this
chapter, we will review the currently available literature
on structures, expression patterns, and potential
functions of both PBAN and PBP in order to gain a
more comprehensive knowledge of pheromone
biosynthesis and reception in Lepidoptera.
Key words: Maruca vitrata, Lepidoptera, Pheromone
biosynthesis activating neuropeptide (PBAN), Pheromone
binding protein (PBP), Phylogenetic relationship
_________________________________________________
*For Correspondence
(email: srini.ramasamy@worldveg.org)

Chapter-19
Application of Nanoparticles in
sustainable Agriculture: Its Current
Status
Atanu Bhattacharyya* 1, Raman Chandrasekar 2,
Asit Kumar Chandra3, Timothy T. Epidi and
Reddy Shetty Prakasham5

and will help in orientation of our agriculture and society

today to new heights in the future; hence it is very much
necessary to nurture the new technology in a sustainable
way.
Key words: Agriculture, nanotechnology, pesticides,
insect pests, food and sustainability, honey bee,
silkworm
____________________________________________
*For Correspondence
email: atanubhatt@rediffmail.com

Nanotechnology Section, Deptarment of Biomedical Engineering,

Rajiv Gandhi Institue of Technology, R.T.Nagar, Hebbal,
Banglore 560 032, India
2
Department of Biochemistry & Molecular Biophysics, Kansas State
University,Manhattan, KS- 66506, USA
3
Central Sericultural Research and Training Institute, Berhampore- 42101,
India
4
Department of Crop Production Technology, Niger Delta University,
Wilberforce Island,P.M.B. 071, Yenagoa, Bayelsa State, Nigeria
5
Bioengineering and Environmental Centre, Indian Institute of Chemical
Technology, Hyderabad - 500 007, India
Abstract

Agriculture is one of the major sectors that provide food

for human, indirectly or directly in addition to feed,
fibre, fire and fuels. The ever increasing population
results in increase of demand of all above tremendously
in spite of constant availability of natural resources. This
is one of the concerning factors to accept the modern
technique, the nanotechnology in particular. This
technology relates with atom by atom arrangement and
helps to produce nanoparticles. The intrinsic properties
of nanoparticles associated with the size, shape and
molecular orientation can react with the agricultural host
tissue suggesting great application potential in the field
of agriculture. Moreover, nanotech based control of
agriculture related diseases also can eliminate by
accumulation of several pesticides from agricultural field
from different sources of host and also use of several
chemical fertilizers which is one of the hot issues for
todays agriculture. This can enhance the quality of life
through its applications in various direction and
dimensions to control the insect pest and the food in
particular. However, the application of this technology
possesses a great deal of difficulty in agriculture sector
but still; it is worth-while at large due to its immense
importance in agriculture application. Moreover, this
new technology is also helpful in the Nano-DNA
Crystals process, Recycling of Agricultural Waste,
Biosensor, Silkworm Industry and Honey bee culture
etc. At present though the application of
nanotechnology in agriculture is in infant stage, this new
technology in all its domains will be discovered gradually

Chapter-20
Mosquito Ribonucleotide Reductase:
A Site for Control
Daphne Q.-D. Pham1, *, Victor H. Perez1, Lissette
Velasquez2, Dharty Bhakta 2, Erica L. Berzin1, Guoli
Zhou2,3, and Joy. J. Winzerling2,3
1

Department of Biological Sciences, University of Wisconsin-Parkside,

Kenosha, WI 53141-2000, USA
2
Department of Nutritional Sciences and 3Center for Insect Science,
The University of Arizona, Tucson, AZ 85721,USA
Abstract

Fidelity in DNA replication is essential to maintain

genomic stability and relies, in part, on ribonucleotide
reductase (RNR). RNR reduces ribonucleotide
diphosphates (rNDPs) to their corresponding deoxy
form (dNDPs), and thereby, provides the rate-limiting
precursors for de novo DNA synthesis. RNR controls the
pools of available dNDPs and responds to
environmental cues that promote cell proliferation.
Work in mammalian systems has shown that altering
RNR expression or activity has profound negative
effects on cell division and integrity. Based on the
mammalian work, it has been hypothesized that similar
results should be seen in insects, specifically mosquitoes.
Thus, work on mosquito RNR was initiated. This review
examines the classification, structure, catalytic
mechanism, and regulation of the yellow fever mosquito,
Aedes aegypti ribonucleotide reductase.
Key words: Aedes aegypti, allosteric inhibitors, mosquito,
ribonucleotide reductase
_________________________________________________
*For Correspondence (email: daphne.pham@uwp.edu )

surfaces to kill insects and vectors with specifically

vectors to human pathogens.
Key words: Silver, Gold, Phytosynthesis, Anopheles
stephensi, Culex quinquefasciatus, Aedes aegypti,
_________________________________________________
*For Correspondence (email: kmvvkg@gmail.com)

Chapter-21
Green protocol for synthesis of metal
nanoparticles to control insect pests
K. Murugan1*, R. Chandrasekar2, C.
Panneerselvam1, A. Naresh Kumar1, P.
Madhiyazhagan1, P. Mahesh Kumar1, Jiang-Shiou
Hwang3, Jiang Wei4
1

Department of Zoology, Bharathiar University, Coimbatore-641046, India

Department of Biochemistry & Molecular Biophysics, Kansas State
University, Manhattan, KS 66506, USA
3
Institute of Marine Biology, National Taiwan Ocean University, Keelung
20224, Taiwan
4
Department of Microbiology, College of Biological Sciences, China
Agricultural University, Beijing 100094, China

Chapter-22
Aquaporins in Blood-Feeding
Arthropods

Abstract

Biosynthesis of metallic nanoparticles is a novel research

area of nanotechnology which has economic and
environmental friendly advantages over conventional
chemical and physical methods of synthesis. Biological
methods were used to synthesize metal nanoparticles of
specific shape and size by enhancing the properties of
nanoparticles in a safer route. Biosynthesis of
nanoparticles may be triggered by several compounds
such as carbonyl groups, terpenoids, phenolics,
flavonones, amines, amides, proteins, pigments, alkaloids
and other reducing agents present in the biological
compounds. Silver (Ag) and gold (Au) nanoparticles
have been the particular focus of plant-based synthesis.
The plant extract based synthesis can provide
nanoparticles of a controlled size and morphology.
Owing to the rich biodiversity of plants in India, their
potential use towards the synthesis of these nobel metal
nanoparticles is yet to be explored. The aim of this
review is to provide the potential to use of nanoparticle

Lisa L. Drake1, Hitoshi Tsujimoto1, Immo A.

Hansen1*
1

Department of Biology, New Mexico State University, Las Cruces,

NM, USA
Abstract

Water movement in and out of cells and tissues is

mediated by a family of water-transporting proteins
named aquaporins (AQPs). Blood feeding insects live on
a water-rich diet and therefore face the challenge to
quickly switch from water conservation to maximal
diuresis after a blood meal. This review focuses on our
current understanding of the AQP family and their
importance for survival and reproduction in bloodfeeding arthropods.
Key words: Aquaporins, aquaglyceroporins, diuresis,
Malpighian tubules, water channel
_________________________________________________
*For Correspondence (email: immoh@nmsu.edu)

effects. The most active of the biostable analogs shows

an LC50 value of 0.0085 nmole/l (LT50 = 1.1 days),
matching or exceeding the potency of some commercial
aphicides. The aphicidal activity of a biostable pyrokinin
can be blocked with an antagonist, indicating the
mechanism occurs via a neuropeptide receptor and
represents a novel and selective mode of action.
Biostable agonists of diapause hormone (a pyrokinin)
can prevent the onset of the protective state of diapause
in the corn earworm, inducing the crop pest to commit a
form of ecological suicide. Alternatively, an antagonist
can block the activity of the native hormone. Biostable
neuropeptide analogs represent important leads in the
development of alternate, environmentally sound pest
insect control agents.
Key words: myotropic, diuresis, biostability, insecticide,
diapause, aphid, agonist, antagonist, corn earworm
_________________________________________________
*For Correspondence (email: nachman@tamu.edu)

Chapter-23
Mimetic analogs of three insect
neuropeptide classes for pest
management
Ronald J. Nachman*
Areawide Pest Management Research, Southern Plains Agricultural
Research Center, USDA, 2881 F/B Road, College Station, TX 77845, USA
Abstract

Neuropeptides are potent regulators of critical life

processes in insects, but are subjected to rapid
degradation by peptidases in the hemolymph (blood),
tissues and gut. This limitation can be overcome via
replacement of peptidase susceptible portions of the
insect neuropeptides to create analogs with enhanced
biostability. Three neuropeptide families, the insect
kinins, insectatachykinins, and pyrokinins stimulate gut
motility and/or Malpighian tubule fluid secretion in
certain insects, but unmodified members demonstrate
little or no effect when fed to pea aphids
(Acyrthosiphon pisum). However, biostable analogs
demonstrate potent oral antifeedant and aphicidal

Chapter-24
Induced resistance in plants and
counter-adaptation by insect pests
Abdul Rashid War and Hari C Sharma*
International Crops Research Institute for the Semi-Arid Tropics
(ICRISAT), Patancheru-502324, Andhra Pradesh-India

Abstract

Insect and plants have coevolved for millions of years.

Plants respond to herbivory through various
morphological, biochemical, and molecular mechanisms
to counter/offset the effects of herbivore attack. These
defense strategies against herbivores are wide-ranging,
highly dynamic, and could be direct and/or indirect.
Direct defense affects the herbivores growth and
development due to antibiosis because of secondary
metabolites produced constitutively and/or upon
infestation by the insect pests. The indirect defense
involves the recruitment of natural enemies of the insect
pests. The natural enemies (parasitoids and predators)
are attracted by the volatiles produced by the plants in
response to damage by the insect pests. The direct and
indirect defensive strategies either act separately or in
conjunction with each other. However, insects have the
ability to adapt to the defensive responses of the plants
through physiological processes, metabolism and
behavior to offset the adverse effects of the host plants
defense systems. This process of defensive responses by
the host plants and counter defense by the insect pests
results in the breakdown of resistance, and evolution of
new populations/biotypes of the insect pests. This coevolution between the plants and insects poses a major
threat for developing crop cultivars with stable
resistance to the target pest for pest management.
Key words: Host plant resistance, induced resistance, insect
adaptation, secondary metabolites, pest management.
_________________________________________________
*For Correspondence (email: H.sharma@cgiar.org )

Chapter-25
Insect Chemical Communication An Important Component of Novel
Approaches in the Insect Pest
Management
Pathipati Usha Rani*
Senior Principal Scientist, Biology and Biotechnology Division, CSIRIndian Institute of Chemical Technology, Hyderabad- 500 007, Andhra
pradesh, India.
Abstract

Communication among insects is an interesting process

that involves several chemicals surrounding them, the
sensory receptors on different appendages with high
precision and finally a congenial environment that acts
as a media for effective communication of the desired
message are necessary for this. The chemicals from
terrestrial plants attract the pest insects towards the host
plant for colonization; whereas, the pest feeding induces
production of different set of chemicals that attract the
natural enemies of the pest towards the pest attacked
plants, thereby to the pest itself. This is a complex, but
interesting as well as important phenomena that is
becoming vital in the field of chemical ecology in recent
years. The role of insect sensory receptors is crucial in
this chemical communication between plants and insects
and almost all the insects are well equipped with suitable
appendages to contribute in this phenomenon. The
morphology of insect sensillae and their role in sensory
perception of plant chemicals is described. Finally the
insect chemical communication has an important role to
play in pest control activities too which is also narrated
herein.
Key words: Chemical Ecology, Sensory receptors,
Chemical communication, Insect - Plant interactions, Plant
defense.
_________________________________________________
*For Correspondence (email:
usharani65@yahoo.com,
usharani65@gmail.com, purani@iict.res.in)

Chapter-26
Mosquito control using biological
larvicides: Current Scenario
Subbiah Poopathi1*, C. Mani1 and Raman
Chandrasekar2

transmitting mosquitoes. In the present study, we focus

the current scenario of biopesticide production from
cost-effective culture media for the control of mosquito
vectors.
Key words: Insecticide, Insect growth regulators, Bacillus
thuringiensis, mosquitoes, malaria, Japanese encephalitis
_________________________________________________
*For Correspondence
(email: Subbiahpoopathi@rediffmail.com )

Unit of Microbiology and Immunology, Vector Control Research Centre

(Indian Council of Medical Research), Medical complex, Indira
Nagar, Puducherry 60 5006, India
2
Department of Biochemistry and Molecular Biophysics, Kansas State
University, Manhattan 66506, KS, USA
Abstract

Emerging infectious diseases are defined as new,

reemerging or drug-resistant infections whose incidence
in humans has increased within the past two decades or
whose incidence threatens to increase in the near future.
The causes of resurgence of infectious diseases include
changes in human industrial practices, economic
development, and changes in land use, increase in
international travel and commerce and adaptation of the
microbes including development of resistance to
antimicrobial agents. Mosquitoes cause great nuisance to
human beings and pose threats to public health as
vectors of diseases like malaria, filariasis, dengue,
Japanese encephalitis, and West Nile fever. Annually 300
million people are estimated to be affected by malaria,
transmitted by Anopheles mosquitoes with more than
one million deaths. The world burden of lymphatic
filariasis is estimated to be 250 million people.
Approximately 20 million people are infected every year
by dengue viruses transmitted by Aedes mosquitoes with
~24,000 deaths. Several strategies have been adopted to
control these dipteran pests and to reduce vector-borne
diseases. Synthetic insecticides have been effectively
used during the past several decades for mosquito
control operations. But the chemical approach has
demerits, such as the development of insecticide
resistance, environmental pollution, bioamplification of
contamination of food chain and harmful effects to
beneficial insects. Hence, there has been an increased
interest in recent years in the use of biological control
agents (Bacillus sphaericus, Bs and Bacillus thuringiensis
subsp. israelensis, Bti) for mosquito control. Though the
high efficacy and specificity of these biological agents are
useful in controlling mosquitoes, the cost to grow the
bacteria through a highly refined laboratory bacterial
culture medium is exorbitant. Hence, efforts are under
process to develop an inexpensive culture media
globally. Considering the abundant supply of these
bioorganic wastes, scientists from all over world have
now utilized these bio-organic waste materials for the
production of biopesticides to control disease

Chapter-27
Application of RNA Interference
toward Insecticide Resistance
Fang Zhu1,2*, Yingjun Cui3, Douglas B. Walsh1,
Laura C. Lavine2
1

Irrigated Agriculture Research and Extension Center, Washington State

University, Prosser, WA, USA
2
Department of Entomology, Washington State University, Pullman, WA,
USA
3
Department of Entomology, University of Kentucky, Lexington, KY, USA

Abstract

Insecticide resistance is a major challenge for effective

and sustainable Integrated Pest Management (IPM).
Recent advances in genomic and post-genomic
approaches not only dramatically improve our
understanding of new mechanisms with regard to
insecticide resistance but also provide potential tactics to
manage pest populations. In this review, we focus on the
latest insights on using RNA interference (RNAi) in
combination with existing strategies to investigate

molecular mechanisms underlying insect adaptation to

insecticides. Importantly, we outline the potential for
RNAi as a new alternative for integrated pest
management strategies
Key words: RNA interference, environmental RNAi,
double-stranded RNA, insecticide resistance, adaptation,
xenobiotic, detoxification, IPM alternative
_________________________________________________
*For Correspondence Email: fang.zhu@wsu.edu (Fang Zhu)

importance. The in-vitro and field assessments to test a

class of pesticide against a pest have become a task for
the researchers. The promising concepts and
methodologies of bioinformatics which are discussed in
detail in this chapter will help agricultural scientists and
entomologists to find the routes to understand the
evolutionary history of pest, its mechanism of action, its
relatedness to other species, etc. This will result in
improvements in the quality of crops, their yield,
controlling the pest to synthesis target oriented drug
design. We also cover certain basic and applied concepts
of bioinformatics in a brief and crispy manner to help
entomologists to grasp them easily. Keeping in mind the
application of these aspects to insects, few worked out
examples are also been given.
Key words: bioinformatics, computational proteomics,
entomo-informatics, ecdysone receptor, homology modeling,
protein databases, phylogenic analysis, molecular docking
________________________________________________
*For Correspondence (email: habeeb.m@ktr.srmunir.ac.in,
biochandrus@yahoo.com )

Chapter-28

Entomo-informatics: A prelude to

the concepts in Bioinformatics

Habeeb, S.K.M.*1, and Raman Chandraskear2

1

Department of Bioinformatics, School of Bioengineering, Faculty of

Engineering & Technology, SRM University, Kattankulathur,
Chennai 603203, Tamilnadu, India
2
Department of Biochemistry and Molecular Biophysics, 238 Burt Hall,
Kansas State University, Manhattan 66506, KS, USA
Abstract

Entomo-informatics actually is as a scientific discipline and

plays an essential role in todays entomological sciences.
The amount of data being generated in the sequencing
labs has resulted in new avenue called bioinformatics.
Currently many researchers in biology are unfamiliar
with available bioinformatics methods, tools and
databases, which could lead to missed opportunities or
misinterpretation of the information. Considering the
rate at which pests have developed resistance to various
insecticides; entomological studies have gained

Chapter-29
Molecular expression and structurefunction relationships of
apolipophorin III in insects with
special reference to innate immunity
Bharat Bhusan Patnaik1,3, Raman Chandrasekar 2,
Yeon Soo Han1*

Chapter-30

Department of Applied Biology, Division of Plant Biotechnology, Institute

of Environmentally-Friendly, Agriculture (IEFA),
College of Agriculture and Life Sciences, Chonnam National University,
Gwangju 500-757, Republic of Korea
2
Department of Biochemistry and Molecular Biophysics, Kansas State
University, Manhattan 66506, KS, USA
3
School of Biotechnology, Trident Academy of Creative Technology
(TACT), Bhubaneswar 751013 Odisha, India
Abstract

Insect immune response relies on the humoral and

cellular mechanisms of innate immunity. The key factors
of the humoral immune mechanisms are the
antimicrobial peptides that act in concert against
invading pathogens. One of the several such key factors
is the apolipophorin III (apoLp-III) gene that is
constitutively expressed in the hemolymph and
functions in immune sensing and activation in insects
against invading pathogenic microbes, apart from its role
in lipid transport. We have reported the functional role
of apoLp-III in the epithelial cells of mosquito gut and
established its action as a negative regulator of
Plasmodium development. Recently, we also studied the
promoter sequence of apoLp-III gene from a
coleopteran beetle, Tenebrio molitor (TmapoLp-III) to
understand the functional regulation imposed by the
putative transcription factors. In addition, the critical
function of TmapoLp-III in immune surveillance against
an intracellular pathogen, Listeria monocytogenes has been
currently explored. Here we review the functions of
apoLp-III in the hemolymph and make an attempt
towards understanding its intracellular role in Anopheles
gambiae apoLp-III (AgapoLp-III). A detailed analysis of
AgapoLp-III sequence and structure has been provided
to dissect the evolution of the protein in insects that
form a basis to authenticate their multidimensional role.
Key words: Anopheles gambiae, apoLp-III, gene
expression, molecular structure, innate immunity
_________________________________________________
*For Correspondence (email: hanys@jnu.ac.kr )

Computer-aided pesticide design: A

short review
Jitrayut Jitonnom*
Division of Chemistry, School of Science, University of Phayao, Phayao
56000, Thailand
Abstract

With a large resistance globally to available insecticides

used today, a new, effective, safer strategy for insect
control is highly demand for agrochemical industry. For
this reason, a rational approach that could be used to
produce more selective and harmless insecticides is
urgently needed. Computer-aided molecular design has
been shown for many years to play essential roles in the
field of insecticide discovery, providing useful
information that could guide the design and
development of new and better insecticidal agents. A
short overview of recent applications of computational
methods on the rational design of insecticides is
presented through several insecticide-targeting receptors,
including acetylcholinesterase, voltage-gated sodium
channel, cytochrome P450 monooxygenase and nicotinic
acetylcholine receptor.
Key
words:
insecticides,
docking,
3D-QSAR,
acetylcholinesterase, voltage-gated sodium channels,
cytochrome P450 monooxygenases, nicotinic acetylcholine
receptor, computer-aided molecular design
*For Correspondence (email: jitrayut.018@gmail.com)

Cover page contributors: Usha Rani, Murugan, K., Raman Chandrasekar, Fang Zhu,
Emmanule J.Joly, Hansen, I., Ada Rafeli, Hiroaki Abe, Michalovia, P.,
Insect Image /
Photograph contributions: Raman Chandrasekar, Geoff Attardo, Murugan, K.,
Sergio Frada, Foto Peti, Yudy Sauw, Hiroaki Abe, John Horstman,
Syed Ishfaq Ali Shah, H.C Sharma, Jill Sterit Murphy, Dennis Hoffman,
Dave Clermont, Juan Carlos GM., Charleen E.M., Marrisia Ruth,
Butterfly Enthusiast Group, Insect photography for fun group,
KSU Insect Zoo, Katalin Hollay, Pamela Judith Wanamaker,
Marc Fischer, Julio Medina
Cover Design:
Production
Manager:
Paper:

Raman Chandrasekar & Brintha

T.R. Govindarajan
Acid free and high quality paper

Printing & Binding: K-State Union, Copy and Printing Services,

Kansas State University, Manhattan 6606, KS, USA.

,
,