Antimicrobial Activities of some Herb and Spices Extracted by Hydrodistillation and Supercritical Fluid Extraction on the Growth of Escherichia coli, Salmonella Typhimurium and Staphylococcus aureus in Microbiological Media


  • Somhak Xainhiaxang National University of Laos
  • Noppol Leksawasdi Chiang Mai University
  • Tri Indrarini Wirjantoro Chiang Mai University


Antimicrobial activity, Herb extracts, Hydrodistillation, Supercritical fluid extraction


This study investigated the antimicrobial actions of Zanthoxylum limonella, neem leaves, garlic and galangal from Laos to inhibit some foodborne pathogens, particularly Escherichia coli, Salmonella enterica serovar. Typhimurium and Staphylococcus aureus. Herb extracts were obtained by hydrodistillation at 100ºC for 4 h at atmospheric pressure or by supercritical fluid extraction at 45ºC and 17 MPa for 4 h. The antimicrobial activities of the extracts were then studied against three different pathogens on microbiological media using Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and agar disc diffusion assay. The highest yield extract was determined in the Z. limonella extract obtained by hydrodistillation, which was 6.32±0.40%. In the MIC method, the Z. limonella extract from hydrodistillation and galangal extract obtained by supercritical fluid extraction at a concentration of 12.5% could inhibit all of the studied pathogens. However, it was only the Z. limonella extract produced by hydrodistillation that could kill the pathogens at the lowest concentration of 12.5%. Regarding the agar disc diffusion assay, Z. limonella extract from hydrodistillation at 100% concentration could inhibit E. coli for 15.67±1.81 mm, which was not significantly different to that of an antibiotic control of 10 mg methicillin (p≥0.05). For S. Typhimurium and Staph. aureus, the highest inhibition could be significantly achieved by galangal extract obtained by supercritical fluid extraction at 100% concentration for 25.87±0.68 and 25.77±0.68 mm, respectively (p<0.05). These inhibition zones were significantly higher than their antibiotic controls of 10 mg amoxycillin and 10 mg methicillin, respectively.


Download data is not yet available.


Almasaudi, S.B., Al-Nahari, A.A.M., Abd El-Ghany, E.S.M., Barbour, E., Al Muhayawi, S.M., Al-Jaouni, S., Azhar, E., Qari, M., Qari, Y.A. and Harakeh, S. 2017. Antimicrobial effect of different types of honey on Staphylococcus aureus. Saudi Journal of Biological Sciences. 24:1255–1261.

Ankri, S. and Mirelman, D. 1999. Antimicrobial properties of allicin from garlic. Microbes and Infection. 2:125–129.

Araújo, S., Silva, I.A.T., Tacão, M., Patinha, C., Alves, A. and Henriques, I. 2017. Characterization of antibiotic resistant and pathogenic Escherichia coli in irrigation water and vegetables in household farms. International Journal of Food Microbiology. 257:192–200.

Bagheri, H., Manap, M.Y.B.A. and Solati, Z. 2014. Antioxidant activity of Piper nigrum L. essential oil extracted by supercritical CO2 extraction and hydro-distillation. Talanta. 121:220–228.

Bajpai, V.K., Baek, K.-H. and Kang, S.C. 2012. Control of Salmonella in foods by using essential oils: A review. Food Research International. 45(2):722–734.

Baker, C.A., Rubinelli, P.M., Park, S.H., Carbonero, F. and Ricke, S.C. 2016. Shiga toxin-producing Escherichia coli in food: Incidence, ecology and detection strategies. Food Control. 59:407–419.

Bakkali, F., Averbeck, S., Averbeck, D. and Idaomar, M. 2008. Biological effects of essential oils - A review. Food and Chemical Toxicology. 46:446–475.

Bubpawan, P., Boonphong, S., Sriwattanawarunyoo, C. and Udeye, V. 2015. Characterization of the essential oil and fatty oil from Makhwaen fruit (Zanthoxylum rhetsa (Roxb.) DC). NU. International Journal of Science. 12(1):1–10.

Charoensup, R., Duangyod, T., Phuneerub, P. and Singharachai, C. 2016. Pharmacognistic specification of Zanthoxylum limonella (Dennst.) Alston: Fruits and seeds in Thailand. Journal of Advanced Pharmaceutical Technology and Research. 7:134–138.

Das, Q., Islam, Md.R., Marcone, M.F., Warriner, K. and Diarra, M.S. 2017. Potential of berry extracts to control foodborne pathogens. Food Control. 73:650–662.

Dušan, F., Marián, S., Katarína, D. and Dobroslava, B. 2006. Essential oils - their antimicrobial activity against Escherichia coli and effect on intestinal cell viability. Toxicology in Vitro. 20:1435–1445.

Fadil, M., Fikri-Benbrahim, K., Rachiq, S., Ihssane, B., Lebrazi, S., Chraibi, M., Haloui, T. and Farah, A. 2018. Combined treatment of Thymus vulgaris L., Rosmarinus officinalis L. and Myrtus communis L. essential oils against Salmonella typhimurium: Optimization of antibacterial activity by mixture design methodology. European Journal of Pharmaceutics and Biopharmaceutics. 126:211–220.

Gupta, A., Mahajan, S. and Sharma, R. 2015. Evaluation of antimicrobial activity of Curcuma longa rhizome extract against Staphylococcus aureus. Biotechnology Reports. 6:51–55.

Gutierrez, J., Barry-Ryan, C. and Bourke, P. 2008. The antimicrobial efficacy of plant essential oil combinations and interactions with food ingredients. International Journal of Food Microbiology. 124:91–97.

Gutiérrez-Morales, A., Velázquez-Ordoñez, V., Khusro, A., Salem, A.Z.M., Estrada-Zúñiga, M.E., Salem, M.Z.M., Valladares-Carranza, B. and Burrola-Aguilar, C. 2017. Anti-staphylococcal properties of Eichhornia crassipes, Pistacia vera and Ziziphys amole leaf extracts: Isolates from cattle and rabbits. Microbial Pathogenesis. 113:181–189.

Hamad, A., Alifah, A., Permadi, A. and Hartanti, D. 2016. Chemical constituents and antibacterial activities of crude extract and essential oils of Alpinia galanga and Zingiber officinale. International Food Research Journal. 23(2):837–841.

Hemeg, H.A. 2018. Molecular characterization of antibiotic resistant Escherichia coli isolates recovered from food samples and outpatient Clinics, KSA. Saudi Journal of Biological Sciences. Available on

Herrero-Fresno, A. and Olsen, J.E. 2018. Salmonella Typhimurium metabolism affects virulence in the host - A mini review. Food Microbiology. 71:98–110.

Lahmar, A., Bedoui, A., Mokdad-Bzeouich, I., Dhaouifi, Z., Kalboussi, Z., Cheraif, I., Ghedira, K. and Chekir-Ghedira, L. 2017. Reversal of resistance in bacteria underlies synergistic effect of essential oils with conventional antibiotics. Microbial Pathogenesis. 106:50–59.

Lins, P. 2018. Antimicrobial activities of spices and herbs against Salmonella Oranienburg. Food Control. 83:123–130.

Mahlangu, Z.P., Botha, F.S., Madoroba, E., Chokoe, K. and Elgorashi, E.E. 2017. Antimicrobial activity of Albizia gummifera (J.F.Gmel.) C.A.Sm leaf extracts against four Salmonella serovars. South African Journal of Botany. 108:132–136.

Maragathavalli, S., Brindha, S., Kaviyarasi, N.S., B. Annadurai, B. and Gangwar, S.K. 2012. Antimicrobial activity in leaf extract of neem (Azadirachta Indica Linn.). International Journal of Science and Nature. 3(1):110–113.

Martínez-Graciá, C., González-Bermúdez, C.A., Cabellero-Valcárcel, A.M., Santaella-Pascual, M. and Frontela-Saseta, C. 2015. Use of herbs and spices for food preservation: Advantages and limitations. Current Opinion in Food Science. 6:38–43.

Mayachiew, P. and Devahastin, S. 2008. Antimicrobial and antioxidant activities of Indian gooseberry and galangal extracts. LWT - Food Science and Technology. 41:1153–1159.

Mejri, J., Abderrabba, M. and Mejri, M. 2010. Chemical composition of the essential oil of Ruta chalepensis L: Influence of drying, hydro-distillation duration and plant parts. Industrial Crops and Products. 32:671–673.

Mongkholkhajornsilp, D., Douglas, S., Douglas, P.L., Elkamel, A., Teppaitoon, W. and Pongamphai, S. 2005. Supercritical CO2 extraction of nimbin from neem seeds - a modelling study. Journal of Food Engineering. 71:331–340.

Negi, J.S., Bisht, V.K., Bhandari, A.K., Singh, P. and Sundriyal, R.C. 2011. Chemical constituents and biological activities of the genus Zanthoxylum: A review. African Journal of Pure and Applied Chemistry. 5(12):412–416.

Oonmetta-aree, J., Suzuki, T., Gasaluck, P. and Eumkeb, G. 2006. Antimicrobial properties and action of galangal (Alpinia galanga Linn.) on Staphylococcus aureus. LWT-Food Science and Technology. 39(10):1214–1220.

Petrović, N.V., Petrović, S.S., Džamić, A.M., Ćirić, A.D., Ristić, M.S., Milovanović, S.L. and Petrović, S.D. 2016. Chemical composition, antioxidant and antimicrobial activity of Thymus praecox supercritical extracts. Journal of Supercritical Fluids. 110:117–125.

Pourmortazavi, S.M. and Hajimirsadeghi, S.S. 2007. Supercritical fluid extraction in plant essential and volatile oil analysis. Journal of Chromatography A. 1163:2–24.

Quelemes, P.V., Perfeito, M.L.G., Guimarães, M.A., dos Santos, R.C., Lima, D.F., Nascimento, C., Silva, M.P.N., Soares, M.J. dos S., Ropke, C.D., Eaton, P., de Moraes, J. and Leite, J.R.S.A. 2015. Effect of neem (Azadirachta indica A. Juss) leaf extract on resistant Staphylococcus aureus biofilm formation and Schistosoma mansoni worms. Journal of Ethnopharmacology. 175:287–294.

Rachkeeree, A., Phianmongkhol, A. and Wirjantoro, T.I. 2014. Individual and combination effects of Thai herb extracts and microwave treatment against Salmonella spp. Food and Applied Bioscience Journal. 2(3):224–240.

Reverchon, E. 1997. Supercritical fluid extraction and fractionation of essential oils and related products. Journal of Supercritical Fluids. 10:1–37.

Reverchon, E. and De Marco, I. 2006. Supercritical fluid extraction and fractionation of natural matter. Journal of Supercritical Fluids. 38:146–166.

Salea, R., Veriansyah, B. and Tjandrawinata, R.R. 2017. Optimization and scale-up process for supercritical fluids extraction of ginger oil from Zingiber officinale var. Amarum. Journal of Supercritical Fluids. 120:285–294.

Samelis, J., Ikeda, J.S. and Sofos, J.N. 2003. Evaluation of the pH-dependent, stationary-phase acid tolerance in Listeria monocytogenes and Salmonella Typhimurium DT104 induced by culturing in media with 1% glucose: A comparative study with Escherichia coli O157: H7. Journal of Applied Microbiology. 95(3):563–575.

Samadi, M., Abidin, Z.Z., Yunus, R., Biak, D.R.A., Yoshida, H. and Lok, E.H. 2017. Assessing the kinetic model of hydro-distillation and chemical composition of Aquilaria malaccensis leaves essential oil. Chinese Journal of Chemical Engineering. 25(2): 216–222.

Santhosha, S.G., Jamuna, P. and Prabhavathi, S.N. 2013. Bioactive components of garlic and their physiological role in health maintenance: A review. Food Bioscience. 3:59–74.

Seydim, A.C. and Sarikus, G. 2006. Antimicrobial activity of whey protein based edible films incorporated with oregano, rosemary and garlic essential oils. Food Research International. 39(5):639–644.

Shylaja, M.R. and Peter, K.V. 2004. The functional role of herbal spices. In K.V. Peter (ed.) Handbook of herbs and spices (pp. 11–21) volume 2. Woodhead Publishing Limited, Cambridge, England.
Shan, B., Cai, Y.-Z., Brooks, J.D. and Corke, H. 2007. The in vitro antibacterial activity of dietary spice and medicinal herb extracts. International Journal of Food Microbiology. 117:112–119.

Stanojević, J.S., Stanojević, L.P., Cvetković, D.J. and Danilović, B.R. 2015. Chemical composition, antioxidant and antimicrobial activity of the turmeric essential oil (Curcuma longa L.). Advanced Technologies. 4(2):19–25.

Subapriya, R. and Nagini, S. 2005. Medicinal properties of neem leaves: A review. Current Medicinal Chemistry – Anti-Cancer Agents. 5:149–156.

Suhaj, M. 2006. Spice antioxidants isolation and their antiradical activity: A review. Journal of Food Composition and Analysis. 19:531–537.

Supabphol, R. and Tangjitjareonkun, J. 2014. Chemical constituents and biological activities of Zanthoxylum limonella (Rutaceae): A review. Tropical Journal of Pharmaceutical Research. 13(12):2119–2130.

Tajkarimi, M.M., Ibrahim, S.A. and Cliver, D.O. 2010. Antimicrobial herb and spice compounds in food. Food Control. 21:1199–1218.

Tangjitjaroenkun, J., Chantarasriwong, O. and Chavasiri, W. 2012a. Chemical constituents of the stems of Zanthoxylum limonella Alston. Phytochemistry Letters. 5(3):443–445.

Tangjitjaroenkun, J., Chavasiri, W., Thunyaharn, S. and Yompakdee, C. 2012b. Bactericidal effects and time-kill studies of the essential oil from the fruits of Zanthoxylum limonella on multi-drug resistant bacteria. Journal of Essential Oil Research. 24(4):363–370.

Tangjitjaroenkun, J., Supabphol, R., Chavasiri, W. 2012c. Antioxidant effect of Zanthoxylum limonella Alston. Journal of Medicinal Plants Research. 6(8):1407–1414.

Turgis, M., Vu, K.D., Dupont, C. and Lacroix, M. 2012. Combined antimicrobial effect of essential oils and bacteriocins against foodborne pathogens and food spoilage bacteria. Food Research International. 48(2):696–702.

Wannissorn, B., Jarikasem, S., Siriwangchai, T. and Thubthimthed. 2005. Antibacterial properties of essential oils from Thai medicinal plants. Fitoterapia. 76:233–236.

Weerakkody, N.S., Caffin, N., Turner, M.S. and Dykes, G.A. 2010. In vitro antimicrobial activity of less-utilized spice and herb extracts against selected food-borne bacteria. Food Control. 21:1408–1414.

Wenqiang, G., Shufen, L., Ruixiang, Y., Shaokun, T. and Can, Q. 2007. Comparison of essential oils of clove buds extracted with supercritical carbon dioxide and other three traditional extraction methods. Food Chemistry. 100:1558–1564.

Wilson, E.A. and Demmig-Adams, B. 2007. Antioxidant, anti-inflammatory and antimicrobial properties of garlic and onions. Nutrition and Food Science. 37(3):178–183.

Wongsrisom, N., Jinata, J., Manosan, B., Kuntakhoo, J., Wankuan, S. and Sriyam, S. 2014. Anti-bacterial activities of essential oils from Mah-Khwuaen (Zanthoxylum limonella Alstron) (in Thai). วารสารวิจัยและพัฒนา มจธ. 37(1):3–15.

Wu, Y., Wang, Y., Li, Z.-H., Wang, C.-F., Wei, J.-Y., Li, X.-L., Wang, P.-J., Zhou, Z.-F., Du, S.-S., Huang, D.-Y. and Deng, Z.-W. 2014. Composition of the essential oil from Alpinia galanga rhizomes and its bioactivity on Lasioderma serricorne. Bulletin of Insectology. 67(2):247–254.

Zhang, Y., Liu, X., Wang, Y., Jiang, P. and Quek, S. 2016. Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus. Food Control. 59:282–289.

Zhao, S. and Zhang, D. 2014. Supercritical CO2 extraction of Eucalytus leaves oil and comparison with soxhlet extraction and hydro-distillation methods. Separation and Purification Technology. 133:443–451.




How to Cite

Xainhiaxang, Somhak, Noppol Leksawasdi, and Tri Indrarini Wirjantoro. 2018. “Antimicrobial Activities of Some Herb and Spices Extracted by Hydrodistillation and Supercritical Fluid Extraction on the Growth of Escherichia Coli, Salmonella Typhimurium and Staphylococcus Aureus in Microbiological Media”. Food and Applied Bioscience Journal 6 (Special):218–239.