In-vitro evaluation of antimicrobial and insect repellent potential of supercritical-carbon dioxide (SCF-CO2) extracts of selected botanicals against stored product pests and foodborne pathogens

  • M. L. Bhavya
  • A. G. S. Chandu
  • Sumithra S. Devi
  • Karl-Werner Quirin
  • Akmal Pasha
  • S. V. N. VijayendraEmail author
Original Article


In the present study, the antimicrobial and the insect repellent activity of 16 botanical extracts obtained by supercritical CO2 (SCF) extraction were evaluated. The present investigation was conducted as there is a necessity for exploration of natural botanical extracts that target both stored product insects and microbes. The antimicrobial activity was studied by disc diffusion and broth microdilution methods against ten microbial species, including Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Listeria monocytogenes), Gram-negative bacteria (Escherichia coli and Salmonella enterica), and fungi (Aspergillus flavus, Aspergillus paraciticus, Aspergillus ochraceous, Aspergillus niger and Penicillium verrucosum). Repellency assay was carried out by area preference method against three coleopteran insects (Tribolium castaneum, Rhyzopertha dominica and Sitophilus oryzae). Among all the extracts, thyme and ajwain were effective against all the tested bacteria with a minimum inhibition concentration (MIC) of 256–1024 µg/mL. Hop extract resulted in better antibacterial activity against all the tested Gram-positive bacteria with a MIC of 32–64 µg/mL. Oregano, thyme and ajwain extracts showed broad-spectrum antifungal activity against all the tested fungi with MIC of 128–1024 µg/mL. Most of the extracts exhibited class V (80.1–100%) repellency against T. castaneum. Extracts of hop, ajwain and thyme were found to have strong repellency against T. castaneum and R. dominica. Therefore, SCF extracts of ajwain and thyme can be explored further for the application of bio-extracts as a growth limiting factors in a microcosm where such consortia thrive.


Foodborne pathogens Coleopteran insects Super critical fluid extraction Insect repellency Microbial inhibition 



The authors thank Director, CSIR-Central Food Technological Research Institute, Mysuru for the support and facilities provided for this study. This research was financially supported by Department of Biotechnology, Government of India (BT/IN/Finnish/06/AP/2013).

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

Supplementary material

13197_2019_4141_MOESM1_ESM.docx (18 kb)
Supplementary material 1 (DOCX 17 kb)


  1. Begaa S, Messaoudi M (2019) Toxicological aspect of some selected medicinal plant samples collected from Djelfa, Algeria Region. Biol Trace Elem Res 187(1):301–306CrossRefGoogle Scholar
  2. Bhavya ML, Chandu AGS, Devi SS (2018) Ocimum tenuiflorum oil, a potential insecticide against rice weevil with anti-acetylcholinesterase activity. Ind Crops Prod 126:434–439CrossRefGoogle Scholar
  3. Bomzan DP, Bhavya ML, Chandu AGS, Manivannan S, Lavanya G, Ramasamy K, Pasha A (2018) Potential of pyrethroid-synergised pyrethrum on stored product insects and implications for use as prophylactic sprays. J Food Sci Technol 55(6):2270–2278CrossRefGoogle Scholar
  4. Chandrasekaran M, Venkatesalu V (2004) Antibacterial and antifungal activity of Syzygium jambolanum seeds. J Ethnopharmacol 91(1):105–108CrossRefGoogle Scholar
  5. Channaiah LH, Subramanyam B, McKinney LJ, Zurek L (2010) Stored-product insects carry antibiotic-resistant and potentially virulent enterococci. FEMS Microbiol Ecol 74(2):464–471CrossRefGoogle Scholar
  6. Cowan MM (1999) Plant products as antimicrobial agents. Clin Microbiol Rev 12(4):564–582CrossRefGoogle Scholar
  7. da Silva RP, Rocha-Santos TA, Duarte AC (2016) Supercritical fluid extraction of bioactive compounds. Trends Analyt. Chem 76:40–51CrossRefGoogle Scholar
  8. Ferry N, Edwards MG, Gatehouse JA, Gatehouse AM (2004) Plant–insect interactions: molecular approaches to insect resistance. Curr Opin Biotechnol 15(2):155–161CrossRefGoogle Scholar
  9. Genena AK, Hense H, Smânia Junior A, Souza SMD (2008) Rosemary (Rosmarinus officinalis): a study of the composition, antioxidant and antimicrobial activities of extracts obtained with supercritical carbon dioxide. Food Sci. Technol. (Campinas) 28(2):463–469CrossRefGoogle Scholar
  10. Gomez-Lopez A, Aberkane A, Petrikkou E, Mellado E, Rodriguez-Tudela JL, Cuenca-Estrella M (2005) Analysis of the influence of Tween concentration, inoculum size, assay medium, and reading time on susceptibility testing of Aspergillus spp. J Clin Microbiol 43(3):1251–1255CrossRefGoogle Scholar
  11. Goyal P, Kaushik P (2011) In vitro evaluation of antibacterial activity of various crude leaf extracts of Indian sacred plant, Ocimum sanctum L. Br Microbiol Res J 1(3):70CrossRefGoogle Scholar
  12. Gutierrez J, Barry-Ryan C, Bourke P (2009) Antimicrobial activity of plant essential oils using food model media: efficacy, synergistic potential and interactions with food components. Food Microbiol 26(2):142–150CrossRefGoogle Scholar
  13. Gyawali R, Ibrahim SA (2014) Natural products as antimicrobial agents. Food Control 46:412–429CrossRefGoogle Scholar
  14. Harein PK, De Las Casas E (1968) Bacteria from granary weevils collected from laboratory colonies and field infestations. J Econ Entomol 61(6):1719–1720CrossRefGoogle Scholar
  15. Hell K, Cardwell KF, Setamou M, Schulthess F (2000) Influence of insect infestation on aflatoxin contamination of stored maize in four agroecological regions in Benin. Afr. Entomol. 8(2):169–177Google Scholar
  16. Hosni K, Hassen I, Chaâbane H, Jemli M, Dallali S, Sebei H, Casabianca H (2013) Enzyme-assisted extraction of essential oils from thyme (Thymus capitatus L.) and rosemary (Rosmarinus officinalis L.): impact on yield, chemical composition and antimicrobial activity. Ind Crops Prod 47:291–299CrossRefGoogle Scholar
  17. Hubert J, Stejskal V, Athanassiou CG, Throne JE (2018) Health hazards associated with arthropod infestation of stored products. Ann Rev Entomol 63:553–573CrossRefGoogle Scholar
  18. Ivanovic J, Misic D, Zizovic I, Ristic M (2012) In vitro control of multiplication of some food-associated bacteria by thyme, rosemary and sage isolates. Food Control 25(1):110–116CrossRefGoogle Scholar
  19. Ivanovic J, Dimitrijevic-Brankovic S, Misic D, Ristic M, Zizovic I (2013) Evaluation and improvement of antioxidant and antibacterial activities of supercritical extracts from clove buds. J Funct Foods 5(1):416–423CrossRefGoogle Scholar
  20. Jilani G, Su HC (1983) Laboratory studies on several plant materials as insect repellants for protection of cereal grains. J Econ Entomol 76(1):154–157CrossRefGoogle Scholar
  21. Karakaya S, El SN, Karagözlü N, Şahin S (2011) Antioxidant and antimicrobial activities of essential oils obtained from oregano (Origanum vulgare ssp. hirtum) by using different extraction methods. J Med Food 14(6):645–652CrossRefGoogle Scholar
  22. Khajeh M, Yamini Y, Sefidkon F, Bahramifar N (2004) Comparison of essential oil composition of Carum copticum obtained by supercritical carbon dioxide extraction and hydrodistillation methods. Food Chem 86(4):587–591CrossRefGoogle Scholar
  23. Larson Z, Subramanyam B, Zurek L, Herrman T (2008) Diversity and antibiotic resistance of enterococci associated with stored-product insects collected from feed mills. J Stored Prod Res 44(2):198–203CrossRefGoogle Scholar
  24. Messaoudi M, Begaa S (2018) Application of INAA technique for analysis of essential trace and toxic elements in medicinal seeds of Carum carvi L. & Foeniculum vulgare Mill. used in Algeria. J Appl Res Med Aromat Plants 9:39–45Google Scholar
  25. Nidhina N, Bhavya ML, Bhaskar N, Muthukumar SP, Murthy PS (2017) Aflatoxin production by Aspergillus flavus in rumen liquor and its implications. Food Control 71:26–31CrossRefGoogle Scholar
  26. Pandey AK, Kumar S (2013) Perspective on plant products as antimicrobial agents: a review. Pharmacologia 4(7):469–480CrossRefGoogle Scholar
  27. Pandey AK, Kumar P, Singh P, Tripathi NN, Bajpai VK (2017) Essential oils: sources of antimicrobials and food preservatives. Front Microbiol 7:2161CrossRefGoogle Scholar
  28. Rohinishree YS, Negi PS (2016) Effect of licorice extract on cell viability, biofilm formation and exotoxin production by Staphylococcus aureus. J Food Sci Technol 53(2):1092–1100CrossRefGoogle Scholar
  29. Rój E, Tadić VM, Mišić D, Žižović I, Arsić I, Dobrzyńska-Inger A, Kostrzewa D (2015) Supercritical carbon dioxide hops extracts with antimicrobial properties. Open Chem. 13(1):1157–1171CrossRefGoogle Scholar
  30. Santoyo S, Cavero S, Jaime L, Ibanez E, Senorans FJ, Reglero G (2005) Chemical composition and antimicrobial activity of Rosmarinus officinalis L. essential oil obtained via supercritical fluid extraction. J Food Prot 68(4):790–795CrossRefGoogle Scholar
  31. Santoyo S, Cavero S, Jaime L, Ibanez E, Senorans FJ, Reglero G (2006) Supercritical carbon dioxide extraction of compounds with antimicrobial activity from Origanum vulgare L.: determination of optimal extraction parameters. J Food Prot 69(2):369–375CrossRefGoogle Scholar
  32. Shokri H, Sharifzadeh A, Khosravi AR (2016) Antifungal activity of the Trachyspermum ammi essential oil on some of the most common fungal pathogens in animals. Iran J Vet Med 10(3):173–180Google Scholar
  33. Sinha AK, Sinha KK (1990) Insect pests, Aspergillus flavus and aflatoxin contamination in stored wheat: a survey at North Bihar (India). J Stored Prod Res 26(4):223–226CrossRefGoogle Scholar
  34. Sovilj MN, Nikolovski BG, Spasojević MĐ (2011) Critical review of supercritical fluid extraction of selected spice plant materials. Maced J Chem Chem Eng 30(2):197–220Google Scholar
  35. Stamenic M, Vulic J, Djilas S, Misic D, Tadic V, Petrovic S, Zizovic I (2014) Free-radical scavenging activity and antibacterial impact of Greek oregano isolates obtained by SFE. Food Chem 165:307–315CrossRefGoogle Scholar
  36. Tauxe RV, Doyle MP, Kuchenmüller T, Schlundt J, Stein CE (2010) Evolving public health approaches to the global challenge of foodborne infections. Int J Food Microbiol 139:S16–S28CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2019

Authors and Affiliations

  1. 1.Food Protectants and Infestation Control DepartmentCSIR-Central Food Technological Research InstituteMysuruIndia
  2. 2.Flavex Naturextrakte GmbHRehlingen-SiersburgGermany
  3. 3.Department of Microbiology and Fermentation TechnologyCSIR-Central Food Technological Research InstituteMysuruIndia

Personalised recommendations