Abstract
Lepidopteran insects are the most dominant and diversified group among phytophagous pests and economically important insects. Gut bacteria has a continuous and dynamic effect on the host gut. In the present investigation, isolation and enumeration of the bacteria from the gut of three lepidopteran insects viz third instar of Antheraea assamensis, fifth instars of Helicoverpa armigera and Plutella xylostella have been carried out to find out their importance in the nutrition of the host by detecting the cellulolytic, amylolytic and lipolytic activity of these bacterial isolates. Screening with biochemical kit (Enterobacteriaceae™ kit) identified 71 gut bacterial isolates which belonged to Enterobacteriales, a dominant group of bacteria inhabiting intestine of insects. A combination of culture dependent assays and qualitative screening has been deployed to detect nutritionally important gut bacterial isolates in the present study with emphasis on screening of lipase, cellulase, amylase and gelatinase producing bacteria based on enzyme activities. Qualitative assays have detected 52 isolates with cellulase, 46 isolates with lipase, 31 isolates with amylase and 2 isolates with gelatinase activities in A. assamensis. Similarly in case of H. armigera, 12 isolates with cellulase, 12 isolates with lipase and 7 isolates with amylase activity were found. In case of P. xylostella 5 isolates with cellulase, lipase, amylase and one with gelatinase activities were found. The study of the enzymatic activities the gut bacteria constitute a starting point to understand the functional significance of these organisms in the metabolism and physiology of the host insect.
Similar content being viewed by others
References
Dillon RJ, Dillon VM (2004) The gut bacteria of insects: nonpathogenic interactions. Annu Rev Entomol 49:71–92
Indiragandhi P, Anandham R et al (2007) Cultivable bacteria associated with larval gut of prothiofos-resistant, prothiofos-susceptible and field caught populations of diamondback moth, Plutella xylostella and their population for, antagonism towards entomopathogenic fungi and host insect nutrition. J Appl Microbiol 103:2664–2675
Visotto LE, Oliveria MGA, Guedes RNC, Ribon AOB, Good-God PIV (2010) Contribution of gut bacteria to digestion and development of the velvet bean caterpillar, Anticarsia gemmatalis. J Insect Physiol 55(3):185–191
Broderick NA, Raffa KF, Goodman RM, Handelsman J (2004) Census of the bacterial community of the gypsy moth larval midgut by using culturing and culture-independent methods. Appl Environ Microbiol 70(1):293–300
Feng W, Wang X-Q, Zhou W, Liu G-Y, Wan Y-J (2011) Isolation and characterization of lipase-producing bacteria in the intestine of the silkworm, Bombyx mori, reared on different forage. J Insect Sci 2:135. http://insectscience.org/II.135
Anand AAP, Vennison SJ, Sankar SG, Prabhu DIG, Vasan PT, Raghuraman T, Geoffrey CJ, Vendan SE (2009) Isoltaion and characterization of bacteria from the gut of Bombyx mori that degrade cellulose, xylan, pectin and starch and their impact on digestion. J Insect Sci 10:107. http://insectscience.org/10.107
Ghose TK (1987) Measuring of cellulase activities. Pure Appl Chem 59:257–268
Huang S, Sheng P, Zhang H (2012) Isolation and identification of cellulolytic bacteria from the gut of Holotrichia parallela larvae (Coleoptera: Scarabaeidae). Int J Mol Sci 13:2563–2577. doi:10.3390/ijms13032563
Cruz TE, Torres JM (2012) Gelatin hydrolysis test protocol. In: ASM, American Society for Microbiology, Microbe Library 1–7
Engel P, Moran NA (2013) The gut microbiota of insects-diversity in structure and function. FEMS Microbiol Rev 37:699–735
Mohanraj P, Subramanian S, Muthuswami M (2009) Survey of gut microflora of select Indian multivoltine and bivoltine breeds of silkworm Bombyx mori L. Sericologia 49(2):201–209
Rogers TE, Doran-Peterson J (2010) Analysis of cellulolytic and hemicellulolytic enzyme activity within the Tipula abdominalis (Diptera: Tipulidae) larval gut and characterization of Crocebacterium ilecola gen. nov. sp. nov., isolated from the Tipula abdominalis larval hindgut. Insect Sci 17:291–302
Delalibera IJ, Handelsman J, Raffa KF (2005) Contrasts in cellulolytic activities of gut microorganisms between the wood borer, Saperda vestita (Coleoptera: Cerambycidae), and the bark beetles, Ipis pini and Dendroctonus frontalis (Coleoptera: Curculionidae). Environ Entomol 34:541–547
Watanabe H, Tokuda G (2010) Cellulolytic systems in insects. Annu Rev Entomol 55:609–632
Oppert C, Klingeman WE, Willis JD, Oppert B, Jurat-Fuentes J (2010) Prospecting for cellulolytic activity in insect digestive fluids. Comp Biochem Physiol B 155:145–154
Mrdakovic M, Lazarevic J, Peric-mataruga V, Ilijin L, Vlahovic M (2009) Partial characterization of a lipase from Gypsy Moth (Lymantria dispar L.) larval midgut. Folia Biol (Krakow) 56:103–110
Terra WR, Ferreira C (1994) Insect digestive enzymes: properties, compartmentalization and function. Comp Biochem Physiol 109B(1):01–62
Park DS, Oh HW, Kim H, Heo SY, Kim N, Seol KY, Park HY (2007) Screening of bacteria producing lipase from insect gut: isolation and characterization of a strain, Burkholderia sp. HY-10 producing lipase. Korean J Appl Entomol 46:131–139
Christeller JT, Amara S, Carriere F (2011) Galactolipase, phospholipase and triacylglycerol lipase activities in the midgut of six species of lepidopteran larvae feeding on different lipid diets. J Insect Physiol 57:1232–1239
Strobl S, Maskos K, Wiegand G et al (1998) A novel strategy for inhibition of α-amylases Yellow meal worm α-amylase in complex with Ragi bifunctional inhibitor at 2.5 Å resolution. Structure 6:911–921
Prasanna VA, Kayalvizhi N, Rameshkumar N, Suganya T, Krishnan M (2014) Characterization of amylase producing Bacillus megaterium from the gut microbiota of silkworm Bomby mori. Res J Chem Environ 18(7):38–45
Darvishzadeh A, Bandani AR, Karimi J, Timouri GH (2012) Biochemical characterization of digestive α-amylase of red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae). Arch Phytopathol Plant Prot 45:2132–2142
Darvishzadeh A, Bandani AR (2012) Identification and characterization of Alpha-amylase in the rose aphid, Macrosiphum rosae (Linnaeus, 1758) (Hemiptera: Aphididae). Munis Zool Entomol 7(2):1089–1096
Leboffe MJ, Pierce BE (2010) Microbiology laboratory theory and application, 3rd edn. Morton Publishing Company, Colorado
Thangamalar A, Ramesh GK, Subramanian S, Mahalingam CA (2009) Use of biochemical kits for characterization of enterobacteriaceae from the gut of silkworm Bombyx mori L. Karnataka J Agric Sci 22:514–516
Acknowledgments
The authors gratefully acknowledge the help and support rendered by ICAR-Indian Agricultural Research Institute, New Delhi and Amity University, Noida, Uttar Pradesh and Central Silk Board-Centre Muga Eri Research and Training Institute, Jorhat, Assam for conducting this study. The financial support rendered by DBT is acknowledged with thanks.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Gandotra, S., Bhuyan, P.M., Gogoi, D.K. et al. Screening of Nutritionally Important Gut Bacteria from the Lepidopteran Insects Through Qualitative Enzyme Assays. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 329–337 (2018). https://doi.org/10.1007/s40011-016-0762-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40011-016-0762-7