Lemongrass ( Cymbopogon) : a review on its structure, properties, applications and recent developments

  • Review Paper
  • Published: 31 July 2018
  • Volume 25 , pages 5455–5477, ( 2018 )

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  • Abu Naser Md Ahsanul Haque 1 ,
  • Rechana Remadevi 1 &
  • Maryam Naebe   ORCID: orcid.org/0000-0002-5266-9246 1  

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This review addresses the structure, properties, applications and future scope of lemongrass, which constitutes an abundant source of plant material around the world. As a source of cellulose, it has been successfully used for the adsorption of metal ions and dyes and for manufacturing paper and pulp. Recently, it has shown promise in the production of composites and bio energy, as well as obtaining silica and other metal oxides. However, previous research studies have mostly concentrated on utilizing the biological activities of the constituents in therapeutic uses, food preservation, cosmetics and agriculture. Therefore, this review covers literature on all areas of current studies on lemongrass and identifies its multidimensional potential. Furthermore, this review describes the intended application of lemongrass as a source of cellulosic matter, more specifically in the materials science field.

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lemongrass research paper pdf

(Drawn by data from Saleem et al. 2003a , b )

lemongrass research paper pdf

(Drawn from the concept of Singh 2014 )

lemongrass research paper pdf

(Redrawn from Singh 2014 )

lemongrass research paper pdf

(Drawn from the concept of Kaur and Dutt 2013 )

lemongrass research paper pdf

(Produced by data from Kamoga et al. 2015 )

lemongrass research paper pdf

(Drawn from the methodology of Bekele et al. 2017 )

lemongrass research paper pdf

(Adapted from Alfa et al. 2014 ; Madhu et al. 2017 )

lemongrass research paper pdf

(Drawn from methodologies of Firdaus et al. 2015 , 2016 )

lemongrass research paper pdf

(Drawn by data from Adeneye and Agbaje 2007 )

lemongrass research paper pdf

(Redrawn from the concept of Ekpenyong and Akpan 2017 )

lemongrass research paper pdf

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We acknowledge the Ph.D. scholarship support from Deakin University for the first author of this paper. Thanks to Dr. Jane Allardyce (Deakin University) for proofreading the draft.

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Haque, A.N.M.A., Remadevi, R. & Naebe, M. Lemongrass ( Cymbopogon) : a review on its structure, properties, applications and recent developments. Cellulose 25 , 5455–5477 (2018). https://doi.org/10.1007/s10570-018-1965-2

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DOI : https://doi.org/10.1007/s10570-018-1965-2

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Lemongrass: a review on its botany, properties, applications and active components

Profile image of Rafia Rehman

Lemon grass (Cymbopogon citratus), is a member of poaceae family. It is a medicinal plant with compounds capable of controlling pathogens and increasing herbal resistance to pathogenic diseases. Lemongrass is widely used in the herbal teas and other non-alcoholic beverages in baked food, and also in the confections. Essential oil from the lemongrass is commonly used as a fragrance in the perfumes and cosmetics, such as creams and soaps. Lemon grass essential oil is comprised of a high content of citral, which is used as a source for the production of beta carotene and vitamin A etc. Hence, due to the presence of various chemical constituents present in lemon grass oil, it uses in different pharmaceutical industries for its anti-depressant, analgesic, antipyretic, bactericidal, anti-septic, carminative and astringent properties.

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Globally, resistance to antibiotics has become a big concern and has a significant effect on the health of patients. Effective treatment approached must be explored in order to resolve this problem. Lemon grass (<em>Cymbopogon citratus</em>) is an essential medicinal plant with numerous biological and pharmacological potential that is helpful towards infectious diseases susceptible to multi-drugs. Therefore, present study was conducted to evaluate the antioxidant potential and antimicrobial activities of different extracts of Lemon Grass. Antioxidant potential of the plant was evaluated by different antioxidant parameters. Multidrug resistant gram positive and negative bacteria were collected from microbiology department. Comparative analysis of lemon grass was performed among aqueous, methanolic and aqueous extracts treated with different temperatures. Study results indicated that highest total phenolic contents were found in lemon grass extract treated at 170ºC while t...

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International Journal of Phytomedicine and Phytotherapy

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Effect of lemongrass ( Cymbopogon citratus Stapf ) tea in a type 2 diabetes rat model

  • Husaina Anchau Garba 1 ,
  • Aminu Mohammed 1 ,
  • Mohammed Auwal Ibrahim 1 &
  • Mohammed Nasir Shuaibu 1  

Clinical Phytoscience volume  6 , Article number:  19 ( 2020 ) Cite this article

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Lemongrass ( Cymbopogon citratus Stapf) tea is a widely consumed beverage for nourishment and the remedy of diabetes mellitus (DM) in Africa locally. The aim of the present study was to investigate the antidiabetic action of lemongrass tea (LGT) in a type 2 diabetes (T2D) model of rats.

The fructose-streptozotocin (STZ) animal model for T2D was used and the LGT was prepared by boiling for 10 min in water, allowed to cool and administered at 0.25 or 0.5% (ad libitum), for 4 weeks to the T2D rats.

The LGT showed higher phytochemical contents compared to the cold-water extract. The diabetic untreated animals exhibited significantly ( p  < 0.05) higher serum glucose and lipids, insulin resistance (HOMA-IR) index with a significantly lower ( p  < 0.05) levels of serum insulin, β-cell function (HOMA-β) and liver glycogen compared to the normal animals. Oral supplemented of LGT for 4 weeks improved these changes comparable to the metformin treated group.

The data suggests that LGT intake had excellent antidiabetic effect in a T2D model of rats attributed to the higher content of the ingredients.

Introduction

Diabetes mellitus (DM) is a metabolic derangement associated with sustained hyperglycemia due to the defective insulin secretion and/or insulin action, resulting to changes in the generation of energy [ 1 ]. Recent data has shown that DM affected over 500 million people globally in 2018, and this number is likely to double by 2045 [ 2 ]. Type 1 (T1D) and type 2 (T2D) are the major classes of DM with T2D being the most prevalent of all diabetes cases. T2D is marked with insulin resistance and partially dysfunctional pancreatic β-cells to adequately secrete insulin in response to hyperglycemia [ 3 ]. Considerable evidence from preclinical and clinical studies has supported the significance of nutraceuticals, functional foods, and dietary patterns in the treatment of T2D [ 4 , 5 , 6 ]. In recent time, diet and food-based therapies for T2D are receiving much attention due to the fear of insulin injection and intake of oral hypoglycemic drugs. In addition, the perceived short- and/or long-term side effects associated with conventional oral hypoglycemic drugs have greatly influence the shift to food-based therapy [ 7 ]. These have prompted the increasing search for potent food-derived ingredients as possible alternative therapies for T2D and Cymbopogon citratus Stapf (Poacae) is among the promising functional food with various medicinal and nutritional potential.

The C. citratus commonly known as lemongrass is native from Asia, Africa, and the Americas, but is widely cultivated in temperate and tropical regions. It is an aromatic grass-like plant, with long slender green leaves and is widely distributed in the tropical and subtropical countries [ 8 ]. For several decades, lemongrass has been reported to be extensively used for a number of folkloric, cosmetic, and nutritional purposes. In Nigeria, Egypt, South Africa and Tanzania, lemongrass tea (LGT) is consumed for the treatment of DM and other related disorders such as hypertension and obesity [ 9 , 10 , 11 , 12 ]. Major active ingredients of LGT were phenolics, flavonoids and terpenoids of which the contents were reported to be higher compared to cold water extract [ 13 ].

In some previous studies aqueous extract of lemongrass was shown to reduce blood glucose and lipid profiles levels in non-diabetic [ 11 , 14 ] and type 1 diabetic rats [ 15 ]. The major limitation to these studies was poor extraction method as most of active ingredients of lemongrass such as citral, limonene and linalool, were volatiles and cold extraction may not properly extract the bioactive compounds. Furthermore, boiling has been the widely method used for the preparation of lemongrass as beverage for nourishment and for the traditional treatment of diabetes across most parts of the world. Interestingly, Bharti et al. [ 16 ] have reported the hypoglycemic, hypolipidemic and antioxidant potential of lemongrass essential oils in hyperlipidemic rats. The main constrains to this study were lack of using T2D animal model and essential oils may not contain some polar component of lemongrass such as the flavonoids. Thus, to the best of our knowledge, the scientific evidence to validate the use of the LGT in the treatment of diabetes, precisely T2D has not been investigated, despite its consumption for medicinal and nutritional purposes. To further support the selection of LGT, Oboh et al. [ 13 ] have reported higher antioxidant property of the LGT than the cold-water extract due to the higher phytochemical content. Therefore, our present study is designed to investigate the antidiabetic potential of the LGT on T2D rat model.

Materials and methods

Plant material.

Fresh lemongrass was collected in January, 2018 from Zaria, Kaduna State, Nigeria. The sample was authenticated at the herbarium unit of the Biological Science Department, Ahmadu Bello University, Zaria, Nigeria and a voucher specimen number 1882 was deposited accordingly. The leaf sample was immediately washed and shade-dried for 2 weeks to constant weight and was ground to a fine powder.

Tea preparation

The lemongrass tea (LGT) preparation was prepared by mimicking the preparation protocols used locally for the treatment of diabetes or consumed for nourishment and was in accordance with Islam [ 17 ] method. Briefly, the fine powdered samples; 0.25 g/100 ml and 0.5 g/100 ml were prepared by boiling for 10 min in water, allowed to cool to room temperature and filtered through Whatmann filter paper (No. 1). The filtrates were supplied to the animals during the 4 weeks intervention period ad libitum. For the phytochemical analysis, the filtrate was evaporated using a rotary evaporator to obtain the extract.

Phytochemical analysis

Flavonoid content.

The method of Bohm and Koupai-Abyazani [ 18 ] was used for the quantification of flavonoids. Briefly, 10 g of the samples were repeatedly extracted in 100 ml of 80% methanol. The mixtures were filtered using Whatman filter paper No. 42 (125 mm). The filtrates were transferred into crucible and evaporated into dryness in water bath and weighed to constant weight.

Total phenolic content

Folin-Ciocalteu method of Chang et al. [ 19 ] was used to quantify phenolic content of the sample. Five grams (5 g) of the samples were boiled in 50 ml of ether for 15 min and then 5 ml of the extracts were pipetted into 50 ml flask. 2 mL of ammonium hydroxide solution and 5 ml of amyl alcohol were also added to the samples and made up to the mark. It was left to react for 30 min for colour development; the absorbance was measured at 550 nm. Gallic acid was used for calibration of a standard curve. The results are expressed as mg gallic acid equivalents (mgGAE)/g dry weight of the plant tissue.

Alkaloid content

The Naili et al. [ 20 ] method was used to determine the alkaloid content. 5 g of the samples were weighed into a 250 ml beaker and 200 mL of 10% acetic acid in ethanol was added and they were covered and allowed to stand for 4 h. The mixtures were filtered and the extracts were concentrated on a water bath to one-quarter of the original volume. Concentrated ammonium hydroxide was added dropwise to each of the extracts until the precipitation was complete. The whole solutions were allowed to settle and the precipitates were collected and washed with dilute ammonium hydroxide and then filtered. The residues were weighed and recorded as alkaloid content.

Saponin content

The method of Obadoni and Ochuko [ 21 ] was used to determine the saponin content. 20 g of the samples were put into a conical flask and 100 mL of 20% aqueous ethanol was added. The mixtures were heated over a hot water bath for 4 h with continuous stirring at about 55 °C. The mixtures were filtered and the residues re-extracted with another 200 mL 20% ethanol. The extract was reduced to 40 mL over water bath at about 90 °C. The concentrates were transferred into a 250 mL separatory funnel and 20 mL of diethyl ether was added and they were shaken vigorously. The aqueous layer was recovered while the ether layer was discarded. The purification process was repeated. A volume of 60 mL of n-butanol was added. The combined n-butanol extracts were washed twice with 10 mL of 5% aqueous sodium chloride. The remaining solutions were heated in a water bath. After evaporation the samples were dried in the oven to constant weight.

Tannin content

The tannin content was assayed according to the AOAC [ 22 ]. Aqueous solutions (25 mL) of the preparations were transferred into1 L conical flask, then 25 ml of indigo solution and 750 ml distilled deionised water were added. 0.1 N aqueous solution of KMnO 4 was used for titration until the blue coloured solution changes to green colour. Then few drops at time until solution becomes golden yellow. Standard solution of Indigo carmine was prepared The blank tests by titration of a mixture of 25 mL Indigocarmine solution and 750 mL distilled water were carried out.

Experimental animals and grouping

Forty-two (42) male Wistar rats were obtained from the Department of Pharmacology, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria with initial mean bw of 211.01 ± 17.05 g. The handling and use of the animals were in accordance with the National Institutes for Health Guide for the Care and Use of Laboratory Animals. The handling and use of the experimental animals were approved by the Animal Research Ethical Committee of the Ahmadu Bello University, Zaria, Nigeria. Animals were housed in transparent cages (3 or 4 animals/cage) with a 12 h light-dark cycle and supplied with a standard rat pellet diet ad libitum during the entire experimental period. The animals were randomly divided into six groups of seven animals namely; NC: Normal Control, DBC: Diabetic Control, DLTL, Diabetic + low dose (0.25%) of LGT, DLTH, Diabetic + high dose (0.5%) of LGT, DMF: Diabetic + metformin (150 mg/kg bw), NLTH: Non-diabetic + high dose (0.5%) of LGT. The animals were allowed to acclimatize for 1 week before starting the experiment. The selection of the dosages used for the LGT and standard metformin were based on trial study and available literature as well [ 23 , 24 ].

Induction of type 2 diabetes (T2D)

In order to induce the two major pathogeneses of T2D, insulin resistance and partial pancreatic β-cell dysfunction, the method of Wilson and Islam [ 25 ] was adopted. Briefly, in the initial first 2 weeks of the experiment, the animals in the DBC, DLTL, DLTH and DMF groups were supplied with a 10% fructose solution (ad libitum) for the induction of insulin resistance while the animals in the NC and NLTH groups were supplied with normal drinking water. After this period and an overnight fast, a low dose of streptozotocin (STZ) at 40 mg/kg bw dissolved in citrate buffer (pH 4.5) were intraperitoneally injected to the animals in the DBC, DLTL, DLTH and DMF groups to induce partial pancreatic β-cell dysfunction, whereas the animals in the NC and NLTH groups were injected with a similar volume of the vehicle buffer. One week after the STZ injection, the fasting blood glucose (FBG) of all animals were measured in the blood collected from the tail vein by using a portable glucometer (Glucoplus Inc., Accucheck). Animals with a FBG level ≥ 200 mg/dl were considered as diabetic [ 26 ] while the animals with a FBG level < 200 mg/dl were excluded from the study.

Intervention period

After the confirmation of diabetes, a respective concentration of the LGT was supplied to the animals for 4 weeks ad libitum in DLTL and DTLH and NLTH groups while the animals in controls (NC and DBC) and DMF groups were treated with similar volume of the vehicle and metformin, respectively. Throughout the experimental period, feed and fluid intake were measured every morning by subtracting the remaining amount of feed and fluid respectively, from the amount given on the previous morning. Moreover, the weekly body weight and FBG levels were measured in all animal groups during the entire intervention period.

Oral glucose tolerance test (OGTT)

To measure the glucose tolerance ability of each animal, the OGTT was performed in the last week of the 4-week intervention period. A single dose of glucose solution (2 g/kg bw) was orally administered to each animal and the subsequent levels of blood glucose were measured at 0 (just before glucose ingestion), 30, 60, 90 and 120 min after the ingestion of glucose.

Collection of blood and organs

At the end of the experimental period, animals were sacrificed by anesthesia and the blood and liver were collected. The whole blood of each animal was preserved immediately after collection and centrifuged at 3000 rpm for 15 min. The serum from each blood sample was separated and preserved in refrigerator for further analysis. The liver was collected from each animal, washed with normal saline, wiped with filter paper, weighed and preserved in a refrigerator until subsequent analysis.

Analytical methods

The serum insulin concentration was measured by an enzyme-linked immunosorbent assay (ELISA) method using an ultrasensitive rat insulin ELISA kit. The serum lipid profile, albumin, total protein concentrations as well as liver function enzymes; aspartate and alanine transaminases (AST and ALT) and alkaline phosphatase (ALP) were measured using commercially available Randox kits, according to the manufactures guide. Homeostatic model assessment (HOMA-IR and HOMA-β) scores were calculated at the end of the intervention period according to the following formula:

LDL-cholesterol was calculated according to Friedewald et al. [ 27 ] equation as shown below:

Liver glycogen concentrations were measured by phenol-sulfuric acid method as described by Good et al. [ 28 ]. Liver tissue (1.0 g) was placed in 4.0 ml of KOH (30%) then heated in boiling water for 10 min. After cooling, 0.2 ml of Na 2 SO 4 (20%) and 5.0 ml of ethanol (95%) were added and kept at 20 °C for 5 min. After precipitation was completed, the mixture was centrifuged at 3000 x g for 10 min and decanted. The packed glycogen in the tube was dissolved by addition of 5.0 ml distilled water with gentle warming. Exactly 5.0 ml of HCl (1.2 mol/L) was added to 5.0 ml of the sample in test tubes then neutralized by the addition of 2 drops of 0.5 mol/L NaOH and a drop of phenol red as indicator then cooled. Glucose in the sample was measured separately. Briefly, a reagent blank was prepared by pipetting 1.0 ml of distilled water into a clean test tube. Exact 1.0 mL of sample and standard glucose solution (0.5 mg/mL of glucose), was pipetted into a similar tube. Then, 5.0 mL of anthrone reagent was delivered into each tube and tightly capped. The test tubes were placed in a cold-water bath. After all tubes have reached the temperature of the cold water, they were immersed in a boiling water bath to a depth little above the level of the liquid in the tubes for 15 min and was then removed and placed in a cold-water bath and cooled to room temperature. Absorbance of test and standard were read against the reagent blank at 620 nm.

Glycogen (mg/g liver tissue) = (Optical density unknown 0.5 x Vol. of sample × 100 × 0.9)/.

Optical density x g of the tissue.

Statistical analysis

All data are presented as the mean ± SD of seven animals. Data were analyzed by using the analysis of variance (ANOVA) (SPSS for Windows, version 22, IBM Corporation, NY, USA) using Tukey’s-HSD multiple range post-hoc test. Values were considered significantly different at p  < 0.05.

Quantitative phytochemical content of LGT

The amount of the phytochemicals presents in the lemongrass tea (LGT) and the cold-water extract is presented in Table  1 . With the exception of saponins, all the ingredients quantified were higher in LGT compared to the cold-water extract, though not statistically ( p  > 0.05) different from each other. However, the amount of total phenolics in LGT was significantly ( p  < 0.05) higher compared to the cold-water extract (Table  1 ).

Effect of LGT on feed and fluid intakes and mean body weight change in T2D rats

The data of the feed and fluid intakes and mean body weight change, are presented in Figs.  1 and 2 , respectively. It was observed that induction of T2D significantly ( p  < 0.05) elevated the feed and fluid intakes and reduced the body weight in the diabetic groups compared to NC. After the 4-week LGT intervention, there was a significant ( p  < 0.05) improvement in the feed and fluid intakes of the treated animals (Fig.  1 ), though the effect on body weight (Fig.  2 ) was not statistically significant ( p  > 0.05) compared to DBC group as well as within the treated diabetic groups. Additionally, there was no significant ( p  > 0.05) effect within the groups treated with the LGT and standard metformin (Figs.  1 and 2 ).

figure 1

Feed and fluid intake in different animal groups during the experimental period. Data are presented as the mean ± SD of 7 animals. a–c Values with different letters over the bars for a given parameter are significantly different from each other group of animals (Tukey’s-HSD multiple range post hoc test, p  < 0.05). NC- Normal control; DBC- Diabetic control; DLTL- Diabetic lemongrass tea low; DLTH- Diabetic lemongrass tea high; DMF- Diabetic metformin; NLTH- Normal lemongrass tea high

figure 2

Mean body weight changes in different animal groups during the experimental period. Data are presented as the mean ± SD of 7 animals. a,c Values with different letters over the bars for a given parameter are significantly different from each other group of animals (Tukey’s-HSD multiple range post hoc test, p  < 0.05). NC- Normal control; DBC- Diabetic control; DLTL- Diabetic lemongrass tea low; DLTH- Diabetic lemongrass tea high; DMF- Diabetic metformin; NLTH- Normal lemongrass tea high

Effect of LGT on weekly blood glucose and oral glucose tolerance test in T2D rats

The result of the weekly blood glucose level is presented in Fig.  3 . From the data, after the induction of T2D, there was a significant ( p  < 0.05) elevation of fasting blood glucose (FBG) level in the DBC group compared to NC group. Consumption of LGT significant ( p  < 0.05) reduced blood glucose level. In the first and second weeks of LGT treatment, there was significantly ( p  < 0.05) reduction of FBG in DLTH compared to DLTL. However, after 4-week intervention, no significant ( p  < 0.05) effect was observed within the LGT treated groups and were comparable to metformin treated group, when no effect was observed in NLTH group (Fig.  3 ). Moreover, it was evident from the results of the OGTT that T2D induction significantly ( p  < 0.05) affected the glucose utilization of the DBC group compared to the NC group (Fig.  4 ). On the other hand, significantly ( p  < 0.05) better glucose tolerance and utilization was observed in the DLTH and DLTL groups compared to the DBC group and was comparable to the DMF group, which again was more prominent in DLTH than DLTL and DMF groups (Fig.  4 ).

figure 3

Weekly blood glucose levels of different animal groups during the experimental period. Data are presented as the mean ± SD of 7 animals. a–c Values with different letters over the bars for a given parameter are significantly different from each other group of animals (Tukey’s-HSD multiple range post hoc test, p  < 0.05). NC- Normal control; DBC- Diabetic control; DLTL- Diabetic lemongrass tea low; DLTH- Diabetic lemongrass tea high; DMF- Diabetic metformin; NLTH- Normal lemongrass tea high

figure 4

OGTT of different animal groups during the experimental period. Data are presented as the mean ± SD of 6 animals. a–c Values with different letters over the bars for a given parameter are significantly different from each other group of animals (Tukey’s-HSD multiple range post hoc test, p  < 0.05). NC- Normal control; DBC- Diabetic control; DLTL- Diabetic lemongrass tea low; DLTH- Diabetic lemongrass tea high; DMF- Diabetic metformin; NLTH- Normal lemongrass tea high

Effect of LGT on serum insulin and the calculated HOMA-IR and HOMA-β indices and other biochemical parameters in T2D rats

The results of serum insulin and the calculated HOMA-IR and HOMA-β indices are presented in Table  2 . According to the data, the serum insulin level and the calculated HOMA-β index were decreased significantly ( p  < 0.05) when HOMA-IR index was increased significantly ( p  < 0.05) in the DBC group compared to the NC group (Table  2 ). After 4-week intervention of LGT, there was significant ( p  < 0.05) and dose-dependent increase in serum insulin levels and the calculated HOMA-β index with a concomitant decrease of HOMA-IR index. These parameters were not altered in the NLTH compared to NC groups (Table  2 ). It was also observed that the liver glycogen content, serum AST and ALP levels were significantly ( p  < 0.05) decreased whereas serum ALT, total proteins and albumin were elevated in the DBC group compared to the NC group (Table  3 ). Treatment with LGT modulated these alterations by reverting to near normal (Table  3 ). In addition, despite no significant ( p  > 0.05) difference was observed within the treated groups in liver glycogen content, serum ALT, AST, ALP, and albumin levels, the effects were more pronounced in DLTH than DLTL groups (Table  3 ).

Effect of LGT on serum lipid profiles in T2D rats

The serum total cholesterol (TC), triglycerides (TG) and low-density lipoprotein (LDL) cholesterol were significantly ( p  < 0.05) decreased in the DBC group compared to the NC group (Fig.  5 ). Consumption of LGT to the diabetic animals significantly ( p  < 0.05) decreased serum TC, TG and LDL-cholesterol levels compared to the DBC group. Moreover, HDL-cholesterol level in DBC group increased but not significantly ( p  < 0.05) different compared to the NC group. Treatment of LGT further reduced HDL-cholesterol, though not statistically significant ( p  < 0.05) compared to the DBC group Additionally, the levels of the above-mentioned parameters were not affected in NLTH group at the end of the intervention period (Fig.  5 ).

figure 5

Lipid profile levels of different animal groups during the experimental period. Data are presented as the mean ± SD of 7 animals. a–c Values with different letters over the bars for a given parameter are significantly different from each other group of animals (Tukey’s-HSD multiple range post hoc test, p  < 0.05). NC- Normal control; DBC- Diabetic control; DLTL- Diabetic lemongrass tea low; DLTH- Diabetic lemongrass tea high; DMF- Diabetic metformin; NLTH- Normal lemongrass tea high

Our present study showed that the LGT consumption for 4 weeks at both dosages reduced blood glucose levels, improved postprandial glucose utilization, ameliorated insulin resistance, hyperlidemia and alterations in some biochemical parameters in T2D rat model. This could be the first report that adequately reported the antidiabetic potential of LGT in T2D rat model. The LGT had higher content of the phytochemicals compared to the cold-water extract, with phenolics having the highest content (Table  1 ). This accord to several previous studies that shows dramatic increase in total phenolics, tannins and alkaloids contents of fruits and vegetables when boiled for less than 30 min than the cold-water extracts, though reason remains speculative [ 29 , 30 , 31 ]. It has been proposed that boiling improves the contact of the phytochemicals with water molecules which subsequently enhances extraction efficiency and in turn greater content of the ingredients in the boiled extract [ 32 ]. Interestingly, Oboh et al. [ 13 ] have shown an increased antioxidant potential of LGT compared to the cold-water extract due to the higher content of the ingredients in LGT. This again supports the selection of LGT for the present study in addition to the widely acclaimed health benefits in the treatment of several diseases including T2D.

Our data showed that induction of T2D decreased the mean body weight, caused polyphagia and polydipsia in the diabetic untreated animals. These, might be linked to the increased energy expenditure, excessive fluid retention and increased eating habit to compensate for the body weight loss in uncontrolled T2D condition [ 33 ]. Consumption of LGT for 4-weeks ameliorated these alterations, signifying possible recovery from the diabetic state. Our results are in line with the previous studies that used cold-water extract [ 11 , 14 ] which have been further supported by the significant reduction in hyperglycemia in LGT treated groups.

Successful reduction of either fasting or postprandial hyperglycemia remains a prime target in the prevention and treatment of T2D and its associated complications [ 34 ]. In the present study, induction of T2D elevated the blood glucose levels and was sustained throughout the study period, indicating the success of the induction. Administration of LGT to the diabetic animals reverted the alteration to near normal (Fig.  3 ). In some previous studies, treatment of lemongrass ethanol and aqueous extracts to non-diabetic animals reduced blood glucose by 21.3% and 19.8%, respectively [ 11 , 14 ]. Similarly, administration of essential oil or aqueous extract reduced blood glucose by about 47.3% and 42.4% in hyperlipidemic and diabetic animals, respectively [ 15 , 16 ]. However, in our present study, consumption of LGT reduced blood glucose by 60.3% indicating greater efficacy, attributed to the higher phytochemical contents compared to other extracts in the literature. Our finding was further supported by higher glucose tolerance ability of the LGT diabetic treated compared to the untreated diabetic rats (Fig.  4 ). This again could partly support the potent inhibitory ability of the lemongrass on the activities of α-amylase and α-glucosidase action reported previously [ 10 ]. Similarly, citral, limonene and linalool, active ingredients of LGT were shown to reduced hyperglycemia and attenuated diabetes-associated complications [ 35 , 36 , 37 ]. Therefore, the antihyperglycemic activity observed in our present study could be linked to the individual or combined action of these phytochemicals.

Studies have shown that chronic uncontrolled T2D may led to reduction of circulating insulin levels, and alter the pancreatic integrity and function [ 38 , 39 ], which were observed in our present study. In addition, the pancreatic β-cell function was greatly tempered with the appearance of insulin resistance in untreated diabetic rats. Consumption of LGT improved the pancreatic integrity via increased serum insulin level, improving pancreatic β-cell function and attenuation of insulin resistance (Table  2 ). This could explain the drastic reduction of the blood glucose level observed as elevated circulating insulin and improved β-cell function facilitated the movement of glucose in to the cells for the energy production.

Consumption of high-fructose diet has been associated with increased hepatic and muscle lipid deposition and accumulation in T2D, via stimulation of lipogenesis [ 40 ]. Previously, oral treatment of lemongrass alcoholic or aqueous extracts (200–1000 mg/kg bw) to non-diabetic, hyperlipidemic and diabetic animals exhibited antihyperlipidemic action [ 11 , 14 , 41 ]. This accord with our present data and further confirms the potential of lemongrass in reducing hyperlipidemia associated with diabetes. The elevated HDL-cholesterol in DBC group although not significantly ( p  > 0.05) different with the NC could be due to the ability of body system to produce more HDL-cholesterol to neutralize the negative effect of higher levels of TC, TG and LDL-cholesterol.

On the other hand, treatment of the lemongrass alcoholic extract for 2 weeks showed no effect on the serum total protein, albumin, ALT, AST, ALP and urea [ 41 ]. However, in our present study, there was reduction of the alterations after LGT administration (Table  3 ), signifying better effect of the LGT compared to the alcoholic extract. This could be attributed to the greater amount of the phytochemicals and the longer study period. The lower glycogen content of the DBC group in our study could be linked to the stimulation of the hepatic glycogen phosphorylase, which apparently may increase glucose output and eventually complicates the hyperglycemia in diabetic condition. Interestingly, treatment of LGT ameliorated the alteration and further shows the ability of the LGT to reverse this alteration associated with diabetes. Phenolics and terpenes which are active ingredients of LGT were shown to increase hepatic glucokinase activity, which augments glucose utilization to promote energy storage in the form of glycogen [ 42 ]. Hence, the increased glycogen content observed in the treated groups could be attributed to the inhibition of glucokinase activity by the major active ingredients. This probably could be the mechanism of antidiabetic effect of LGT.

In conclusion, oral intervention of LGT demonstrated antidiabetic actions via improving body weight gain, reducing food and fluid intake and hyperglycemia, improving glucose tolerance ability, insulin sensitivity, β-cell functions and dyslipidemia in T2D model of rats. Hence, our findings suggest that consumption of LGT may provide a good management option for T2D patients with no considerable side effects which also supports the antidiabetic claims of the tea. Further clinical study is required to confirm the effects in human subjects and specific active ingredient responsible for the observed action.

Availability of data and materials

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

Alkaline phosphatase

Alanine transaminase

Aspartate transaminase

Diabetic control

Diabetic metformin

Diabetic lemongrass tea high dose

Diabetic lemongrass tea low dose

Enzyme-linked immunosorbent assay

Homeostatic model assessment-β cell function

Homeostatic model assessment-insulin resistance

Lemongrass tea

Normal control

Non-fasting blood glucose

Normal lemongrass tea high dose

Oral glucose tolerance test

  • Type 2 diabetes

Streptozotocin

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The authors thank Mal. Mua’zu Mahmud for his technical assistance during the study.

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Husaina Anchau Garba, Aminu Mohammed, Mohammed Auwal Ibrahim & Mohammed Nasir Shuaibu

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H.A Garba performed experiments and provided equipment and re-agents; A. Mohammed conducted the statistical analysis and wrote the manuscript; M. A Ibrahim and M. N Shuaibu made manuscript revisions. The authors read and approved the final manuscript.

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Garba, H.A., Mohammed, A., Ibrahim, M.A. et al. Effect of lemongrass ( Cymbopogon citratus Stapf ) tea in a type 2 diabetes rat model. Clin Phytosci 6 , 19 (2020). https://doi.org/10.1186/s40816-020-00167-y

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Language: English | French

The repellency of lemongrass oil against stable flies, tested using video tracking

Activité répulsive de l’huile essentielle de citronnelle contre les stomoxes, testée par tracking vidéo, frédéric baldacchino.

1 Dynamique et Gouvernance des Systèmes Écologiques, Centre d’Écologie Fonctionnelle et Évolutive (CEFE), UMR 5175, Université Paul-Valéry (UM3) Montpellier France

Coline Tramut

2 Laboratoire de Parasitologie, École Nationale Vétérinaire (ENVT) Toulouse France

Emmanuel Liénard

Emilie delétré.

3 Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), UR-Hortsys Montpellier France

Michel Franc

Thibaud martin, gérard duvallet, pierre jay-robert.

Lemongrass oil ( Cymbopogon citratus ) is an effective repellent against mosquitoes (Diptera: Culicidae) and house flies (Diptera: Muscidae). In this study, its effectiveness was assessed on stable flies (Diptera: Muscidae) in laboratory conditions. First, we demonstrated that lemongrass oil is an active substance for antennal olfactory receptor cells of Stomoxys calcitrans as indicated by a significant increase in the electroantennogram responses to increasing doses of lemongrass oil. Feeding-choice tests in a flight cage with stable flies having access to two blood-soaked sanitary pads, one of which was treated with lemongrass oil, showed that stable flies ( n  = 24) spent significantly more time in the untreated zone (median value = 218.4 s) than in the treated zone (median value = 63.7 s). No stable flies fed on the treated pad, whereas nine fed on the untreated pad. These results suggest that lemongrass oil could be used as an effective repellent against stable flies. Additional studies to confirm its spatial repellent and feeding deterrent effects are warranted.

L’huile essentielle de Cymbopogon citratus est un répulsif actif contre les moustiques (Diptera : Culicidae) et les mouches domestiques (Diptera : Muscidae). Dans cette étude, nous avons testé son efficacité contre les stomoxes (Diptera : Muscidae) en laboratoire. Nous avons tout d’abord démontré par électroantennographie (EAG) que l’huile essentielle de C. citratus était une substance active sur les récepteurs olfactifs des antennes de Stomoxys calcitrans , par la mise en évidence d’une augmentation significative des réponses EAG à des doses croissantes d’huile essentielle. Des tests de choix réalisés en cage de vol avec des stomoxes ayant à disposition deux supports imprégnés de sang, l’un ayant été traité avec de l’huile essentielle, montrent que les stomoxes ( n  = 24) ont passé significativement plus de temps dans la zone non traitée (valeur médiane = 218,4 s) que dans la zone traitée (valeur médiane = 63,7 s). Aucun stomoxe ne s’est nourri sur le support traité alors que neuf stomoxes se sont nourris sur le support non traité. Ces résultats suggèrent que l’huile essentielle de C. citratus pourrait être utilisée comme répulsif contre les stomoxes. Des études complémentaires sont nécessaires pour confirmer ses effets répulsifs et anti-gorgement.

Introduction

The stable fly Stomoxys calcitrans L. is among the most damaging arthropod pest of livestock worldwide [ 8 , 15 , 23 ], with a high economic impact on dairy and beef cattle production [ 3 , 27 , 39 ]. It is also a potential mechanical vector of animal pathogens such as equine infectious anemia virus, Trypanosoma evansi , and Besnoitia besnoiti [ 7 , 9 , 19 ]. Control of stable fly populations includes various methods, such as chemical control (pesticides and repellents), cultural control (sanitation), mechanical control (trapping devices), and biological control (parasitoids and entomopathogenic fungi) [ 9 , 20 ]. The best approach is the simultaneous use of several methods in an integrated pest-management program [ 26 ]. Management of adult flies is accomplished mainly with topical insecticides, applied directly to animals. However, continued or repeated use of conventional insecticides often results in the development of resistance and fosters serious human health and environmental concerns [ 13 , 42 ]. Populations of S. calcitrans resistant to pyrethroids and/or organophosphates have already been described in North America and in Europe [ 4 , 21 , 31 , 34 ]. As a result, there have been increased research efforts for natural and environmentally friendly repellents, particularly those based on essential oils [ 38 ]. Several plant-based repellents, such as citronella oil, eucalyptus oil, catnip oil, and zanthoxylum oil, have previously been tested against stable flies and have shown a reduction in attraction and in feeding [ 1 , 13 , 40 , 43 ]. These repellents can be applied topically on animals or in livestock barns [ 13 ]. The first study demonstrating the potential application of a plant-based repellent was conducted by Zhu et al. [ 44 ], in which wax-based catnip pellets spread in the manure/soil areas of cattle feedlots resulted in over 99% repellency of stable flies.

Lemongrass oil is the essential oil obtained from the aerial parts of Cymbopogon citratus (DC.) Stapf., Poaceae [ 29 ]. Geranial ( α -citral) and neral ( β -citral) are the two main active components of lemongrass oil, but other compounds, such as geraniol and citronellol, which are known repellents, are also present in small amounts [ 2 , 18 , 38 ]. Lemongrass essential oil has previously shown a repellent effect, alone or in combination, against different species of disease-transmitting mosquitoes (Diptera: Culicidae) and the house fly Musca domestica L. (Diptera: Muscidae) [ 16 , 25 , 30 , 37 ], and is already present in commercially available products [ 5 , 32 ]. Therefore, our objectives were to verify the sensitivity of antennal receptor cells of S. calcitrans to lemongrass oil and to evaluate its repellency against stable flies using a video-tracking system.

Materials and methods

Stomoxys calcitrans pupae were obtained from the laboratory colony of the National Veterinary School of Toulouse (Toulouse, France) [ 35 ]. Newly emerged flies were not sexed. Males and females were enclosed together in a cotton mesh cage (40 cm W × 25 cm H × 25 cm D) at 24 ± 2 °C with 40–50% relative humidity. Flies were fed with 10% sugar water ad libitum and, once a day, with citrated bovine blood. Experiments were conducted with 2–4-day-old flies. Flies were not fed for 24 h prior to each test.

Electroantennogram recording

Following the method used in the study by Jeanbourquin and Guerin [ 14 ], electroantennogram (EAG) recordings from antennae of S. calcitrans were made with an EAG recording device (EAG combi probe internal gain ×10, CS-55 stimulus controller and IDAC-2 signal acquisition controller, Syntech, Hilversum, the Netherlands). Recordings were made using electrolyte-filled (0.1 M KCl) glass capillary electrodes (Ø 1.5 mm, 40 mm L), with Ag/AgCl wire (Ø 0.5 mm, 20 mm L) making contact with the recording apparatus. The antenna was maintained in a humidified charcoal-filtered air stream delivered at 14.6 mL/s through a metal tube. Aliquots of pure lemongrass oil (from C. citratus DC., citral ~75%, Sigma Aldrich Chemie GmbH, Buchs, Switzerland) were prepared using hexane (95%, Carlo Erba Reagenti, Arese, Italy) at 0.1, 0.01, 0.001, 0.0001 mg/μL. Tested solutions (10 μL) were deposited on a strip of filter paper (20 × 5 mm) placed in a glass Pasteur pipette. The solvent was allowed to evaporate for 15 min before first use. The tip of the pipette was connected to the metal tube, and the test stimulus was delivered to the antenna using an air pulse (20 mL/s for 0.6 s). Stimuli were released successively in random order at 90-s intervals to avoid receptor saturation. Octenol (1-octen-3-ol, 98%, Sigma Aldrich Chemie GmbH, Buchs, Switzerland) was used as a positive control and hexane was used as a negative control. Differences in EAG responses were evaluated using a Wilcoxon signed-rank test.

To observe the flight behavior of stable flies, we used a screen cage (30 cm W × 15 cm H × 15 cm D) made of polyester mosquito netting suspended on a metal frame ( Figure 1 ). A small hole in the middle of one side of the cage was sealed with a piece of cotton wool and was used to allow the release of one fly at a time into the cage. The cage was surrounded by a shield of white foam board to prevent optical stimulation of the flies. To stimulate the fly to move in the cage, pieces of blue and black fabric (SuperMaine 300 g cotton/polyester 65/35%; TDV industries, Laval, France), commonly used to attract biting flies, were hung on each side of the foam board [ 17 , 24 ]. Illumination was provided by fluorescent tubes (frequency 50 Hz) placed below and above the screen cage. The light level in the middle of the cage was about 4600–5000 lux.

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Aerial view of the system for spatial repellency bioassays. The screen cage was made of mosquito netting suspended on a metal frame and surrounded by white foam board with blue and black fabric on each side. Two blood-soaked sanitary pads were set under the cage: one, impregnated with lemongrass oil, was placed in the treated zone, and the other, impregnated with hexane, was placed in the untreated zone. For further details see Materials and Methods section.

One fly was released into the cage 15 min before the test. Bioassays were conducted using male and female stable flies during the daytime at ambient laboratory temperatures of 22–26 °C and 40–50% relative humidity. The bioassays consisted of feeding-choice tests in which the fly had access to two blood sources, one of which was treated with lemongrass oil. Citrated bovine blood (1.5 mL), previously heated at 45 °C, was placed on two sanitary pads (Ø 4 cm) from which we removed the outer layer. The outer layer of one pad was impregnated with 100 μL of lemongrass oil solution at 0.1 mg/μL, and the other outer layer with 100 μL of hexane. When the solvent had evaporated, each outer layer was repositioned on top of one of the blood-soaked sanitary pads, which were placed just under the cage floor, 20 cm apart. Fly movement was recorded using a Digital Video Camera Recorder (DCR-SR21E; Sony, Japan) set 1 m above the center of the cage. The behavior of the fly was then recorded during a 10-min period. We tested 4–6 flies each day; the behavior of 24 flies was included in this study. The room was ventilated for at least 30 min between each test, and a new screen cage was used for successive flies. The positions of the pad treated with lemongrass oil and the untreated pad were inversed each time. The cages were cleaned every day by soaking them in a 2% solution of Decon 90 (Decon Laboratories Limited, Sussex, England) for 12 h.

Video analysis

The video records of fly movement were analyzed using EthoVison XT (v. 8.0; Noldus Information Technology, Wageningen, the Netherlands) [ 28 ]. The cage was defined as an arena (30 × 15 cm) divided into three zones (each 10 × 15 cm): untreated, intermediate, and treated ( Figure 1 ). Movement was recorded at 25 video frames per second, and the fly was tracked by dynamic subtraction ( Figure 2 ). In this method, the program compares each sampled image with a reference image that is updated regularly. Image processing algorithms are applied to detect the fly against the background and to extract relevant image features. During data acquisition, EthoVision displays the live video image, tracking statistics (elapsed time, number of samples), and the x , y co-ordinates of the fly [ 28 ]. Several parameters were calculated: the distance moved (in centimeters), the total time spent in each zone (in seconds), the time spent in movement (in seconds), and the mean velocity (centimeters per second). “Moving” and “not moving” were defined with thresholds at 1 and 0.9 cm/s. A comparison between males and females was made with the non-parametric Mann-Whitney test for independent samples. Comparisons of flight parameters between the treated zone and the untreated zone were made with the non-parametric Wilcoxon signed-rank test for two samples of univariate data. All analyses were performed using PAST version 2.12 [ 12 ].

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Track showing the 10-min recording of a stable fly in the bioassay cage divided into three zones: the untreated zone, the intermediate zone, and the treated zone.

Lemongrass oil volatiles

To estimate the diffusion of lemongrass oil volatiles in the bioassay cage, we compared the atmospheric concentrations of neral and geranial, its most abundant constituents. To accomplish this, three 65 μm Polydimethylsiloxane-Divinylbenzene (PDMS-DVB) fibers (Supelco, Sigma-Aldrich, Bellefonte, PA, USA) were conditioned in the inlet of a gas chromatograph (GC) held at 250 °C for 5 min before sampling. The SPME holders were exposed in the cage for 10 min at three positions. One SPME fiber was positioned 10 cm above each of the two blood-soaked sanitary pads, and another was positioned in the middle of the cage. Relative concentrations of volatile samples were analyzed in a GC-mass spectrometry (MS; Shimadzu QP2010plus, Shimadzu Scientific Instruments, Kyoto, Japan), using helium as the carrier gas (1 mL/min). Samples were injected in splitless mode. The temperature program for GC analyses was 40 °C for 5 min, 5 °C/min to 220 °C, and 10 °C/min to 250 °C.

Results and discussion

Our investigation showed that S. calcitrans EAG amplitudes increased significantly in a dose-dependent fashion with increasing doses of lemongrass oil in the stimulus pipette. The mean EAG amplitude elicited by each dose (0.001 mg: 2.06 ± 0.37 mV; 0.01 mg: 3.37 ± 0.47 mV; 0.1 mg: 5.80 ± 0.67 mV; 1 mg: 6.50 ± 0.57 mV) was significantly greater than that elicited by hexane (1.46 ± 0.29 mV) ( Figure 3 ) and there was no significant difference between lemongrass oil and the octenol at 1 mg on filter paper (6.64 ± 0.55 mV). Octenol is a very strong chemostimulant for S. calcitrans antennae [ 36 , 41 ] and a good attractant in the field [ 11 ]. The study by Zhu et al. [ 44 ] was the first to report that stable fly antennae are also capable of detecting repellents such as catnip oil. In our study, EAG responses to lemongrass oil at 10 μg (~3350 μV) were nearly five times higher than the EAG responses to the same amount of catnip oil (~700 μV recorded by Zhu et al. [ 44 ]). These results indicate that lemongrass oil is a strong stimulant for the olfactory receptor cells of S. calcitrans and thus a suitable candidate for behavioral tests.

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Mean relative EAG amplitudes recorded from Stomoxys calcitrans antennae ( n  = 7) stimulated with lemongrass essential oil at doses of 0.001 mg, 0.01 mg, 0.1 mg, and 1 mg. Hexane was used as negative control. EAG amplitudes are relative to the value of 100% for octenol at 1 mg in the stimulus syringe. Differences in EAG amplitudes were evaluated using the Wilcoxon signed-rank test. Significant differences are indicated by different letters ( p  ≤ 0.05).

In the bioassays, the amount of lemongrass oil on the treated pad used in all tests was 10 mg. Relative concentrations of neral and geranial in the arena were assessed by the height of their peaks in mass chromatograms to reveal a 12-fold decrease in the atmospheric concentration of lemongrass oil between the treated and untreated pads. It should be noted, however, that this measurement was taken in the absence of a fly. The air flow induced by the flight activity of a fly in the cage might partially disturb this ratio during a test.

We tested 11 males and 13 females in the bioassay cage ( Table 1 ). First, we compared the flight activity between the two sexes. The distance moved is considered to be the main indicator of the activity level of a fly [ 22 ]. At the beginning, females were more active than males (in terms of time spent in movement and velocity) ( Figure 4 ). Over the duration of the 10-min recordings, the distance moved by females gradually decreased to reach a level similar to males. This decrease in movement might have been due to exposure to lemongrass oil, or simply to acclimation to the bioassay cage. This is an open question as no tests were conducted without a treated pad. However, locomotor activity was sexually distinct, as has been observed in fruit flies, Drosophila melanogaster [ 10 ].

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Time-course (mean ±  SE ) of the distance moved by stable fly males and females for 10 min following release into a bioassay cage. Curves represent the distance moved for each successive 1-min period during 10-min recordings.

Comparison of the flight activity of male and female stable flies (Mann-Whitney U test), and comparison of the behavior of flies (both sexes) between the zone treated with lemongrass oil and the untreated zone (Wilcoxon W signed-rank test).

Data that show significant differences are indicated in bold.

Comparing the behavior of stable flies in the zone treated with lemongrass oil with their behavior in the untreated zone did not reveal any significant differences between the two zones in terms of the time spent in movement or in the mean velocity of movement ( Table 1 ). However, stable flies spent significantly more time flying in the untreated zone than in the treated zone during the tests. Moreover, we observed nine stable flies feeding on the untreated pad, whereas none fed on the treated pad. The attractiveness of the untreated blood-soaked pad versus the treated pad explains the difference in the total time spent between the two zones. These findings suggest that lemongrass oil could be used as a repellent against stable flies. However, further investigations on spatial repellency and feeding deterrence are necessary to demonstrate that lemongrass oil is as effective as catnip oil against stable flies in the field [ 44 ]. Video tracking appears to be a useful tool to study insect behavior in response to repellent volatiles [ 6 , 33 ], especially for flies, which are otherwise difficult to track.

Acknowledgments

We would like to thank the École Nationale Vétérinaire (ENVT) of Toulouse for providing us with S. calcitrans pupae and CSIRO-CIRAD for access to their EthoVision XT system. We are very grateful to Bruno Buatois, CEFE-CNRS, for his technical assistance. All electroantennogram recordings and GCMS analyses were acquired at the CEFE-CNRS – Platform of Chemical Analysis in Ecology of the LabEx CeMEB, “Centre Méditerranéen de l’Environnement et de la Biodiversité”, 1919 route de Mende, 34293 Montpellier Cedex 5, France.

Cite this article as : Baldacchino F, Tramut C, Salem A, Liénard E, Delétré E, Franc M, Martin T, Duvallet G & Jay-Robert P: The repellency of lemongrass oil against stable flies, tested using video tracking. Parasite, 2013, 20 , 21.

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