Acta agriculturae Slovenica, 118/1, 1–11, Ljubljana 2022 doi:10.14720/aas.2022.118.1.2212 Original research article / izvirni znanstveni članek Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae Scopoli, 1763 (Homoptera: Aphididae) Nadia BOUABIDA 1, Karima BENOUFELLA–KITOUS 1, 2, Samia AIT AMAR 1, Ferroudja MEDJDOUB- BENSAAD 3 and Farid GRAICHE 4 Received May 20, 2021; accepted January 24, 2022. Delo je prispelo 20. maja 2021, sprejeto 24. januarja 2022 1 Laboratory of Production, Improvement and Protection of Plants, Department of Animal and Plant Biology, Faculty of Biological Sciences and Agronomic Sciences, Mouloud Mammeri University of Tizi-Ouzou, Tizi-Ouzou, Algeria 2 Corresponding author, e-mail: ben.kitous@yahoo.fr 3 Laboratory of Production, Safeguard of Threatened Species and Crops, Department of Animal and Plant Biology, Faculty of Biological Sciences and Agronomic Sci- ences, Mouloud Mammeri University of Tizi-Ouzou, Tizi-Ouzou, Algeria 4 Department of mathematics, Faculty of sciences, Mouloud Mammeri University of Tizi-Ouzou, Tizi-Ouzou, Algeria Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae Scopoli, 1763 (Homoptera: Aphididae) Abstract: The objective of our work was the valorisation of four aromatic plants growing spontaneously in Kabylia (Al- geria) by the evaluation of their insecticidal activity against the black bean aphid Aphis fabae Scopoli, 1763. These are oregano, thyme, rosemary and lavender. The plants were subjected to a phytochemical analysis to determine their secondary metabo- lites composition. The results obtained showed that the four extracts are toxic to A. fabae and can significantly reduce its populations. LD50s of 7.76 %; 8.91 %; 9.72 % and 12.88 % were recorded for extracts of oregano, rosemary, thyme and lavender respectively. In addition, the phytochemical screening shows the presence of flavonoids, tannins, saponins and polyphenols. The polyphenols extraction indicates that the oregano extract is the richest with a polyphenol content of 218.73 (± 0.22) µg GAE ml-1. This substance has significant biocidal power. Key words: Aphis fabae; mortality rate; Lamiaceae; poly- phenols; phytochemical screening Ovrednotenje biocidne aktivnosti izvlečkov listov štirih vrst ustnatic (Lamiaceae) na črno fižolovo uš, Aphis fabae Scopoli, 1763 (Homoptera: Aphididae) Izvleček: Predmet razikave je bilo ovrednotenje insekti- cidne aktivnosti štirih aromatičnih rastlin, ki rastejo samoniklo na območju Kabylije (Alžirija) na črno fižolovo uš Aphis fabae Scopoli, 1763. Izbrane rastline so bile dobra misel, timijan, rož- marin in sivka. V rastlinah so s fitokemičnimi analizami določi- li sestavo sekundarnih metabolitov. Rezultati so pokazali, da so bili vsi štirje izvlečki strupeni za črno fižolovo uš in so znatno zmanjšali njeno populacijo. Vrednosti LD50 so za izvlečke do- bre misli, rožmarina, timijana in sivke znašale 7,76 %; 8,91 %; 9,72 % in 12,88 %. Fitokemične analize rastlin so pokazale pri- sotnost flavonoidov, taninov, saponinov in drugih polifenolov. Izvleček polifenolov je pokazal, da je bila na njih najbogatejša dobra misel z vsebnostjo 218,73 ( ± 0,22) µg GAE ml-1, kar kaže na njeno znatno biocidno moč. Ključne besede: Aphis fabae; smrtnost; Lamiaceae; poli- fenoli; fitokemijska analiza Acta agriculturae Slovenica, 118/1 – 20222 N. BOUABIDA et al. 1 INTRODUCTION Herbivorous insects are responsible for significant losses to agriculture due to food damage, but also by car- rying pathogens such as viruses (Kortbeek et al., 2019). They have implemented a range of strategies allowing them to discover their host plants and then reproduce and develop at their expense (Huignard, 2013). Aphids are considered among the most important pests of crops. They cause significant financial losses, and are responsi- ble for yield losses in many cultivated plants (Tagu et al., 2004). These pests cause direct damage by sucking sap, and indirectly by transmitting phytopatogenic viruses and by secreting honeydew on which sooty mold is deposited, which decreases photosynthesis (Blackman and Eastop, 2000; 2006). The use of insecticides is one of the first methods used against aphids (Esmaieli-Vardanjani et al., 2013). However, as a result of excessive use of these prod- ucts, some aphids have developed resistance to most of them. In addition, insecticides destroy beneficial organ- isms and disrupt ecosystems (Harmel et al., 2010; Sabahi et al., 2011). It is therefore necessary to have an alternative con- trol program and to search for new products, on the one hand, to ensure effective protection of agricultural pro- duction, and on the other hand, to contribute to sustain- able management of the environment (Bouchelta et al., 2005). For this purpose, several control methods are rec- ommended to limit the level of aphid outbreaks in crops (Harbaoui et al., 2008). Among these, the use of plant extracts endowed with insecticidal activities which of- fer a certain potential (Isman, 2000). In fact, plants have developed different mechanisms to defend themselves against insects, including the production of specialized metabolites that act as natural insecticides (Kortbeek et al., 2019). According to Huignard (2013), plants are rich in phenolic compounds. The latter are of significant physio- logical and morphological importance in the plant king- dom; they are involved in growth, reproduction and give the plant some resistance against pathogens and pests (Maroun et al., 2013). The main objective behind this work is to study the insecticidal activity of four aqueous extracts of Lamiaceae: oregano (Origanum floribundum Munby), thyme (Thymus numidicus Poir.), rosemary (Rosmarinus officinalis L.) and lavender (Lavandula stoe- chas L.) on the black bean aphid Aphis fabae Scopoli, 1793 (Hemiptera: Aphididae) which can transmit 30 vi- ral diseases to plants (Jahan et al., 2019), as well as the research of the main active elements and the content of polyphenols that exist in each plant. 2 MATERIALS AND METHODS 2.1 HARVESTING PLANT MATERIAL During this experiment, four species of Lamiaceae were tested on the black bean aphid, these are rosemary (Rosmarinus officinalis L.), thyme (Thymus numidicus Poir.), oregano (Origanum floribundum Munby) and lav- ender (Lavandula stoechas L.). Rosemary and lavender were harvested in a private garden located in the region of Draâ El Mizan at an altitude of 432  m (36° 32’8’’N. 3° 50’3’’E.). Whereas the thyme came from the moun- tains of Ait Bouadou (Ouadia) at an altitude of 828 m (36° 30’0’’N. 4° 1’0’’E.), and oregano was harvested in a private plot located in the Makouda region at an altitude of 458 m (36° 47’27’’N. 4° 4’1’’E.). 2.2 PREPARATIONS OF AQUEOUS EXTRACTS For each tested plant, leaves are dried and then crushed. 50 g of powder for each plant were macerated in 1 l of distilled water for 24 hours. Resulting solutions were filtered then stored in vials and kept in the dark. 11 doses (1 %, 2 %, 3 %, 4 %, 5 %, 10 %, 20 %, 30 %, 40 %, 50 % and 100 %) whose concentrations vary according to the dilution of the stock solution in the distilled water were prepared. 2.3 EXPERIMENTAL DESIGN An experimental design formed from 144 jars of in- fested beans are treated with the four plant extracts by means of contact. 12 batches including a control batch have been used for each extract. Aphids were given one dose per batch, with three repetitions for each dose. The three control jars were treated with distilled water. 2.4 APPLICATION OF TREATMENTS After sowing the bean seeds (‘Histal’) in plastic pots, small seedlings obtained were infested with 40 aphids; larvae of different stages and adults. After three days of infestation, the treatment was applied by spraying dif- ferent doses of the four aqueous solutions on the aphid colonies. In order to follow the chronological evolution of aphids mortality subjected to different extracts at dif- ferent concentrations, observations were made daily after spraying. A magnifying glass has been used to count the dead and alive aphids. Acta agriculturae Slovenica, 118/1 – 2022 3 Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae ... 2.5 CHARACTERIZATION OF MAIN CHEMICAL COMPONENTS Phytochemical study enables the detection of bioac- tive secondary metabolites existing in the leaves of the plants tested. It is based on coloring and precipitation re- actions by specific chemical reagents. The method used is the approach adopted by Tona et al. (1998) and Longaga et al. (2000). Besides, total polyphenol content is obtained by spectrophotometry according to Folin-Ciocalteu (FCR) method (Singleton et al., 1999). 2.6 DATA ANALYSIS Mortalities in the treated batches (Mo) were ex- pressed according to Abbott’s formula (1925) in cor- rected mortality (Mc), taking into account natural mor- talities observed in the control batches (Mt). Mortality is expressed as a percentage calculated using the following formula: Mc = (Mo-Mt / 100-Mt) x 100 Mc: Corrected mortality rate Mo: Mortality rate in the treated population Mt: Mortality rate in the control population 2.7 KRUSKAL-WALLIS TEST The Kruskal-Wallis non-parametric test has been used to compare the average mortalities obtained with different treatments. It is based on classification in as- cending order of all observations (Legras and Kohler, 2007). 2.8 CALCULATION OF LETHAL DOSE (LD50) AND LETHAL TIME (TL50) The method of Finney (1971) based on the regres- sion of the probits of mortalities depending on the loga- rithms of the doses of solutions tested made it possible to determine the LD50. The 50 % lethal dose of each extract was estimated, after exposure of aphids to different con- centrations tested. These values were determined from an experimental curve giving the variations in mortality according to increasing concentrations of the extracts. The 50 % lethal time (TL50) was also determined by the same method. The time is transformed into a logarithm and the percentage of mortality corrected into probit. 3 RESULTS After spraying the leaf solutions of the four Lami- aceae on A. fabae populations, an increase in the per- centage of mortality of the latter appeared as a function of time and dose (Fig. 1). Total mortality was observed after 12 days of treatment at the highest doses (50 % and 100 %) for oregano, thyme and rosemary extracts, and at the 100 % dose for lavender extract (Table 1). 3.1 KRUSKAL-WALLIS TEST The mortality-dose boxplot shows that the mor- tality rate is dose dependent; as the dose increases, the mortality rate increases (Fig. 2). This is confirmed by the Kruskal Wallis test, with a p-value of 5.834e-14 for thyme, 3.891e-16 for oregano, 2.695e-12 for rosemary and 9.585e-14 for lavender. These values are less than 0.05. This means that there is a highly significant effect of dose on aphid mortality. The Mortality-time boxplot shows that the mortal- ity rate is higher at time T6 = 12 days after treatment (Fig. 3). This is confirmed by the Kruskal Wallis test with a p-value of 1.811e-14 for thyme, 9.912e-14 for oregano, 5.711e-16 for rosemary and 5.006e-16 for lavender. These values confirm that there is a very significant effect of time on aphid mortality. 3.2 DETERMINATION OF THE LD50 OF THE VARIOUS TREATMENTS TESTED ON A. FABAE After spraying the aqueous extracts of the four plants tested on different batches of aphids, at doses ranging from 1 % to 100 %, the percentages of mortal- ity transformed into probits were recorded and plotted in figure 4. The LD50 of each product confirms the re- sults obtained in the tests. Indeed, the LD50 obtained showed that the solutions tested present a high degree of toxicity towards these insects. Oregano extract gave the lowest lethal dose (7.76 %). It is therefore the most toxic extract compared to the other biopesticides with LD50s of 8.91 %, 9.72 % and 12.88 % for rosemary, thyme and lavender extracts respectively. 3.3 DETERMINATION OF THE TL50 OF THE VAR- IOUS TREATMENTS TESTED ON A. FABAE Calculation of the lethal time showed that the ex- tracts tested on the black bean aphid had a fairly high Acta agriculturae Slovenica, 118/1 – 20224 N. BOUABIDA et al. biocidal activity that could reduce 50 % of the pest popu- lation on the 5th day after spraying the solutions based of oregano leaves (123.03 hours) and of thyme (128.82 hours) and after 6 days for those based on rosemary leaves (134.89 hours) and lavender (141.25 hours) leaf solutions (Fig. 5). Figure 1: Aphid mortality rate (a: Thyme extract, b: Oregano extract, c: Rosemary extract, d: Lavender extract) Dose (%) Corrected mean Mortality (%) Oregano Thyme Rosemary Lavender 1 3.03 0.88 2.58 1.76 2 5.61 2.95 4.13 5.46 3 20.7 12.61 15.51 7.81 4 26.1 33.63 33.57 32.86 5 38.42 35.66 37.34 38.57 10 65.79 51.83 47.68 44.93 20 66.53 60.64 52.25 48.09 30 70.29 61.81 53.44 64.81 40 76.06 82.25 69.18 65.94 50 100 100 100 87.29 100 100 100 100 100 Table 1: Corrected mean mortality of populations of Aphis fabae after 12 days of treatment with the 4 aqueous extracts Thyme Oregano Rosemary Lavender Total tannins + + + + Catechetical tannins + + + + Gallic tannins + + + + Anthocyanins ˗ ˗ + ˗ Saponoside + + + + Mucilages + + + + Reducing sugars + + + + Flavonoids + + + + Total Polyphenols +++ +++ +++ +++ Glucosides + + + + Starch + + + + Protein + + + + Table 2: Results of the chemical analysis of the four plants tested (-): absence of substance; (+): presence of substance; (+++): very high substance content Acta agriculturae Slovenica, 118/1 – 2022 5 Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae ... Figure 2: Mortality-dose boxplot for the four aqueous extracts (a: Oregano extract, b: Thyme extract, c: Rosemary extract, d: Lavender extract) Figure 3: Mortality-Time boxplot for the four aqueous extracts (a: Oregano extract, b: Thyme extract, c: Rosemary extract, d: Lavender extract) 6 Acta agriculturae Slovenica, 118/1 – 2022 N. BOUABIDA et al. Figure 4: Aphid mortality rate as a function of the logarithm of the dose (a: Oregano extract, b: Thyme extract, c: Rosemary extract, d: Lavender extract) Figure 5: Aphid mortality rate as a function of the logarithm of time (a: Oregano extract, b: Thyme extract, c: Rosemary extract, d: Lavender extract) 7Acta agriculturae Slovenica, 118/1 – 2022 Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae ... 7 3.4 CHEMICAL COMPOSITION OF THE EX- TRACTS TESTED The results of the phytochemical characterization tests for the four species made it possible to demonstrate the presence of several common compounds between them. These are polyphenols, flavonoids, tannins, sapon- osides, mucilges, reducing sugars, glucosides, starch and proteins (Table 2). The presence of anthocyanins was only observed in Rosmarinus officinalis. Our results also showed that all four plants have very high polyphenol content. 3.5 TOTAL PHENOL CONTENT UV/Visible spectrophotometry made it possible to quantify the level of polyphenols present in the extracts prepared from the four plants. The results obtained are expressed in microgram equivalents of gallic acid stand- ard used per ml of extract (µg  GAE ml-1 extract) and determined by the equation: y = ax + b. The results are reported in Figure 6. The results obtained revealed that the four plants are rich in polyphenols with a total phenol content that var- ies between 64.2 µg GAE ml-1 and 218.73 µg GAE ml-1. From these results, it appears that oregano contains the highest concentration of polyphenols compared to the other Lamiaceae species. 4 DISCUSSION Under the conditions of this study, aqueous extracts obtained from Lamiaceae leaves had an effect on the mortality of A. fabae populations. The results obtained after the bioassays with the 4 biopesticides showed a di- rect relationship between aphid mortality rates on the one hand and product concentration and exposure time on the other hand. Aphids were very sensitive to the bioassays as extract concentrations increased and time passed. In the range of eleven concentrations tested, the 50 % and 100 % doses induced high toxicity on aphids after 12 days of contact. The reference control caused low mortality rates com- pared to the biopesticides tested. Several studies have shown that the toxic effect of plant extracts is related to the concentration of the ex- tract and the period of exposure. The work of Habou et al. (2011) showed that the bi- ocidal effect of the essential oil of Jatropha Jatropha cur- cas L. on the black bean aphid increases with the increase of the dose. They obtained a mortality rate of 100 % for the 15 % dose after 96 h. Authors indicate that the num- ber of dead aphids increases with the duration of treat- Figure 6: Polyphenol content of the four Lamiaceae expressed in micrograms of gallic acid equivalents per milliliter of extract (µg GAE ml-1) 8 Acta agriculturae Slovenica, 118/1 – 2022 N. BOUABIDA et al. ment, it is after 72 hours that they observe a high mortal- ity rate and which becomes stable between 96 and 120 hours. Besides, Akantetou et al. (2011) mentioned that the mortality rate of aphids subjected to different concen- trations of oil of the whitish basil (Ocimum canum Sims) increased numerically in a linear manner according to the observation periods (1, 3, 5 and 24 hours). Similarly, Laznik et al. (2011) stated that the highest mortality rate of Aphis pomi populations occurred after the third day after treatment with Symphytum officinale and Calendula officinalis extract compared to the first and second day post-treatment. The insecticide effect-dose relationship was also confirmed by the study conducted by Benoufella-Kitous et al. (2014) on A. fabae. According to these authors, the two aqueous extracts of nettle and fern showed a consid- erable insecticidal effect at the highest dose, i.e. 73.8 % and 75.1 % mortality respectively. The biocidal effect of nettle and fern on A. fabae increased during the days fol- lowing the application of the aqueous extracts, reaching a maximum after 6 days. Our results are confirmed by those found by Baba-Aissa et al. (2017) who showed that the toxicity of the essential oil of sour orange is depend- ent on the dose, indicating that more the dose increases, more the formulation presents a greater biocidal effect which results in a reduction in the density of popula- tions of A. fabae. In addition, Kulimushi’s study (2014) showed that the degree of toxicity of the extracts is re- lated to the dose used. After treatment of populations of A. fabae with aqueous extracts of garlic and papaya leaves and their combination, the average number of aphids de- creased from 1 to 36 aphids and on average 86.92 % of aphids were controlled. These results are in agreement with those Saïfi and Belhamra (2018) noted that the toxicity of the es- sential oil of Thymus pallescens de Noë depends on the concentration. These authors recorded on populations of A. fabae, after 24 h, a mortality rate of 34.75 % at the dose of 12 µl ml-1. In a study on the insecticidal activity of sage against the black bean aphid, Benoufella-kitous et al. (2020) demonstrated that the effect of this plant with regard to this pest is the highest 9 days after treat- ment. Furthermore, Oulebsir-Mohand Kaci et al. (2015) reported that the mortality rate of Myzus persicae (Sulzer, 1767) treated with two plant extracts Eucalyptus globulus Labill. and Thymus vulgaris L. increases proportionally with increasing dose. The latter state that the D4 dose (8 µl ml-1) shows a mortality rate of 65.4 % and 71.7 % re- spectively for the two extracts and that the aphid mortal- ity rate is increasing over time. Likewise, in a study on the biological activity of santolina (Santolina africana Jord. & Fourr) against the aphid Aphis craccivora C.L.Koch, 1854[, Lebbal et al. (2017) demonstrated that the toxic effect of this plant with regard to this pest was proportio- nal to the dose, with a mortality rate of 80 % at the dose of 15  % after 24 hours. According to Kumar and Patel (2017), the extract of Cassia angustifolia M.Vahl. showed a toxic effect on Brevicoryne brassicae (L., 1758) with a mortality rate of 100 % at doses of 7 % and 10 % after 72 hours of exposure. On the other hand, Acheuk et al. (2017) mentioned that the crude ethanolic extract of Artemisia judaica L. revealed potent insecticidal effects against the black aphids, A. fabae. Total mortality (100 %) was reached 2 hours after treatment with the highest concentration. Be- sides, Lebbal et al. (2018) found that the extract obtained by maceration of Thymus algeriensis Bioss. & Reut. at a concentration of 25 % was the most effective, with a lar- val mortality rate of 70 % of A. fabae individuals after 24 h. The toxic effects of the solutions could depend on their chemical composition and the sensitivity level of the insects. According to Saidj (2007), among the plants whose efficacy has been evaluated, aromatic plants of the Labiatae family were the most active as direct insecticides but also as inhibitors of oviposition and larval develop- ment of insects. Chiasson and Beloin (2007) suggested that biopesticides act directly on the cuticle of insects and mites, especially soft-bodied ones such as aphids. The phytochemical study of the four aqueous ex- tracts showed that these plants contain mainly flavo- noids, tannins, saponosides and other polyphenols. These results confirm that plants of the Lamiaceae family are medicinal plants rich in secondary compounds that have a toxic effect against insect pests. Similarly, Asghari et al. (2017), in a phytochemical analysis study carried out on several plants of Lamiaceae used in medicine in Aligudarz region, in Iran, show that the three species of thyme: Thymus daenesis Celak, Thymus eriocalyx (Ron- niger) Jalas, and Thymus lancifolius Celak are rich in fla- vonoids and tannins. According to Huignard (2013), secondary com- pounds in plants can cause the death of insects that try to consume them by disrupting the functioning of the nerv- ous system, the digestive system or by preventing larval growth. For instance, phenolic compounds are toxic when ingested by phytophagous insects (Kortbeek et al., 2019). They are present in all parts of higher plants: roots, stems, leaves, flowers, fruits and seeds (Medic-Saric et al., 2003; Boizot and Charpentier, 2006). According to Gale- otti et al. (2008), secondary metabolites such as alkaloids, saponosides, polyphenols (flavonoids and tannins) have pharmacological and toxicological activity. Some alka- loids, anthocyanins, flavonoids, quinines, lignans, ster- oids, and terprnoids have commercial application in the pharmaceutical and biomedical fields and are included in 9Acta agriculturae Slovenica, 118/1 – 2022 Evaluation of biocidal activity of four Lamiaceae leaves on the black bean aphid Aphis fabae ... drugs, dyes, flavours, fragrances and insecticides (Teix- eira Da Silva, 2004). The results obtained revealed that the four plants are rich in polyphenols with a total phenol content var- ied between 64.2 µg GAE ml-1 and 218.73 µg GAE ml-1. These results of the quantitative analysis of polyphenols are close to those of several authors. Celiktas et al. (2007) noted polyphenol concentrations for the crude extract of Rosmarinus officinalis ranging from 34.1 to 119 mg GAE g-1. Cocan et al. (2018), in a study on the biological ac- tivity of rosemary officinale extracts, state that this plant has a polyphenol content that is 86.05 ± 0.40 mg GAE g-1. Fadili et al. (2015) mentioned that Rosmarinus offici- nalis and Thymus satureioides Coss. are rich in polyphe- nols for all the fractions studied and the concentration of polyphenols varies between 21.66 ± 2mg GAE g-1 to 185.71 ± 4 mg GAE g-1. Kholkhal et al. (2013) reported that the polyphenol content of Thymus ciliatus Desf. is 64.23 mg GAE g-1. In Morocco, Bachiri et al. (2016) noted that Lavandula stoechas and L. dentata L. are composed of polyphenol contents of 150.34 mg g-1 and 184.02 mg g-1 respectively. In general, oregano extract was the most effective (LD50 = 7.76 %, TL50 = 123.03 hours) on aphid popula- tions. Low LD50 values indicate strong insecticidal activ- ity and low TL50 values indicate a rapid biocidal effect. The obtained results seem to show that the polyphenols with the highest concentration in this plant would be the active ingredient that plays a determining role in the bi- ocidal activity of this plant. The richness of the aqueous extract in chemically active compounds could explain the traditional use of this plant in various fields. How- ever, the synergistic additive effect of the different com- pounds may also be a factor explaining the remarkable activity revealed by the rosemary extract. The latter was found to be highly toxic to aphids (LD 50 = 8.91 %). Lavender extract ranked last (LD50 = 12.88 %, TL50 = 141.25 hours) compared to the other solutions tested despite its high polyphenol content. This would suggest that the insecticidal activity of plants is not limited to some of their major constituents; it could also be due to some minority constituents. According to Akantetou et al. (2011), the difference in toxicity between the different extracts could be explained by the growing conditions, the harvesting period and the climatic and edaphic con- ditions. The distribution of secondary metabolites may change during plant growth. This may be related to con- ditions of high temperature, sun exposure, drought and salinity, which stimulate the biosynthesis of secondary metabolites such as polyphenols (Falleh et al., 2008; Za- ouali et al., 2010). Some works has shown that the toxic effect of ex- tracts depends on the nature of the plant. According to Khalfi-Habes and Sellami (2010), oregano shows a stronger insecticidal action than rosemary and thyme. These results confirm those obtained in the present study. Similarly, Kumar and Patel (2017) state that Curcuma angustifolia Roxb. causes a higher toxic effect compared to the other plants tested (Cercis gigantean L., Cannabis sativa L., Parthenuim hysterophorus L., Lobelia chinensis Lour., Solanum nigrum L. and Ageratum conyzoides L.). In a study on the biological activity of nettle and fern on the black bean aphid, Benoufella-Kitous et al. (2014) showed that the most significant toxic effect was record- ed for the second species. Benoufella-Kitous (2015) notes that among 8 plant species tested against A. fabae, the most toxic extracts were those from the leaves of toothed lavender with an efficacy of 99.4 % at the 10 % dose, sage with an efficacy of 98.5 % at the 40 % dose and garlic with an efficacy of 97.9 % at the 10 % dose. 5 CONCLUSION The present study showed the relative importance of the use of botanical pesticides, namely aqueous extracts of oregano, rosemary, thyme and lavender leaves, against the black bean aphid. 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