The Citronella and Savory Essential Oils Effect on Honeybees (Apis mellifera L.) Mortality

The Citronella and Savory Essential Oils Effect on Honeybees (Apis mellifera L.) 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Mortality Khalila BENGOUGA, Farida BETTICHE, Souad TAHARCHAOUCHE, Haroun FADLAOUI, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3843753/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 10 Sep, 2025 Read the published version in International Journal of Tropical Insect Science → Version 1 posted 5 You are reading this latest preprint version Abstract Recently a trend to use small concentrations of the essential oils of ( Cymbopogonschoenanthus ) or ( Saccocalyxsatureioides ) to control pests has extended. They are often used to suppress biotic enemies such as fungus, moulds, bacteria and insects. What is not known is whether these oils also influence beneficial beings such as honeybees. In preliminary experiments foragers’ bees were exposed by inhalation to different concentrations of C. schoenanthus ) or ( S. satureioides essential oils. Results showed that bees’ mortality in inhalation test increased with increase in concentrations of essential oils and exposure time. Both essential oils showed LC 50 greater than 5 µl/ml.Meanwhile 205 µl/ml concentrations were more detrimental. Disturbance of locomotion and contractions of bees recorded by video sequences registered revealed that bees exposed to C. schoenanthus presented lesser contractions and were more active than bees exposed to S. satureioides whom presented more contractions and lesser locomotion activity. Our research provides further evidence supporting the use of conditioning techniques to evaluate the use of essential oils as optimal substances to be considered as alternative controls of pests assuming honey bee safety. Essential oils Toxicity Honey bee Figures Figure 1 Figure 2 Background Insecticides are used on different crops to control their insect-pests. Along with harmfulinsects these insecticides also kill the beneficial insects especially pollinators like honey bees (Husain et al. 2014 ).The tremendous increase in crop yields associated with the ‘green’ revolution has been possible in part by the discovery and utilization of chemicals for pest control. However, concerns over the potential impact of pesticides on human health and the environment has led to the introduction of new pesticide registration procedures, such as the Food Quality Protection Act in the United States. These new regulations have reduced the number of synthetic pesticides available in agriculture. Therefore, the current paradigm of relying almost exclusively on chemicals for pest control may need to be reconsidered. New pesticides, including natural product-based pesticides are being discovered and developed to replace the compounds lost due to the new registration requirements (Dayan et al. 2009 ). For that recourse to natural derived insect control agents such as essential oils is proposed as apromising alternative to protect beneficial insects. As their effectiveness on pathogen agents was proved especially of some plants essential oils such: basil, colocynth, jasmine, geranium, senna, chamomileas (Sharaby et al. 2020 ),aloe, ginger, garlic, and hot pepper (Siam and Othman 2020 ), neem (Gopal et al. 2021 ). To prove the antibactirial and fungicidal activities of the essential oils of Cymbopogonschoenanthus (Poaceae) and Saccocalyxsatureioides (Lamiaceae); different studies were maintained in Algeria. High efficacy against E. coli, an average efficacyagainst Staphylococcusaureus and a limited efficacy against Pseudomonasaerogenosof S.satureioides E O was reported byLahrech ( 2010 ). It was reported also as having a broad spectrum of activity as it inhibits both the growth of bacteria that moulds and yeasts. (Ariech and Hendel 2013 ). The results obtained by Sari et al. ( 2013 ) indicate that the E O of Saccocalyx possesses an intermediate anti-leishmania activity in vitro. Fungicidal activity of C.schoenanthus E O is also reported by Amraoui(2014).As well Guerrah&Segueni (2015)demonstrated methanolic extract of C.schoenanthus , antimicrobial activity against E.coli. Meanwhile insecticidal activity of C. schoenanthus emulsified oil can validly be used as alternative in management of P. Xylostella larvaeon cabbages (Laba2012). Results on the Effect of insecticide formulation based essential oil of C.schoenanthus (L.)aplicated on cotton in Togo showed a significant reduction of cotton pests Aphis gossypii Glover (Homoptera :Aphididae) population and an improvement in the production in accordance with the increase of the concentration of C. schoenanthus essential oil (Bokobana et al.2016).For now in Algeria Aous et al. ( 2019 ) studied antioxidant, antimicrobial and insecticidal properties of this plant specieds. Meanwhile the effects of both plants extracted oils to honeybees have been briefly researchedthus,Fuselli et al. (2010) have tested citronella grass ( C.nardus ) and palmarosa ( C.martinii ) from Gramineae to potentially control Paenibacillus larvae, the causal agent of American foulbrood disease (AFB). In order to promote local aromatic plants in the fight against crop pests and reduce the use of synthetic chemical pesticides and the risks associated with them, the effectiveness of an insecticide formulation based essential oil was elaborated by a project proposed by CRSTRA aiming to control Tutaabsoluta on tomatoes and Aphis gossipy on melon; thus two essential oils of two plants C.schoenanthus (Poaceae) and S.satureioides (Lamiaceae) were exploited. But as all other fight methods, the use of pesticide plants presents some favours as well as some limits (Yarou et al. 2017 ). Our subject is whether this option does not have any effect on beneficial insects especially on pollinators such as Apismellifera . Methods The aim of the present study was to evaluate toxic inhalation of C. schoenanthus and S. satureioides to bees’ workers. Vegetal material Plants of both species C. schoenanthus and S. satureioides were collected from Djelfa and Bousaadasteppic regions of Algeria during April 2016 and 2017. The plants were dried up for a period of one month in aired obscure place. Every two days plants were verified and turned to avoid mould. 100 g dried leaves of both plants were subjected to hydro-distillation by a Clevenger device of 1000 ml flask for 4 hours at 100°C.Extracted essential oil was diluted using distilled water. Bees Adult honeybee workers of Apismellifera L. were collected from the bees’ hives from Bio-Ressources station of CRSTRA located at El-Outaya. The hives at the time the bees were collected were free of diseases. No hive treatments to control diseases were conducted before collection of bees. Honeybees were collected from the flight board at the hive entrance at morning time of the day of the experiment, (OEPP 2010) and putted into a plastic container and were brought to laboratory. Inhalation Test Treatments used in the bioassays are the EO extracted of both plants where solutions of 5, 10, 15 and 20 µl/ml are prepared in distilled water on the same day just before the test. The surface residual method in glass jars (Radwan and Taha 2012) with some modifications is adopted for testing contact toxicity of different concentrations of E Os to honeybees ( A. mellifera ).Within plasticboxes(Detzeland Wink 1993) of 15x20x30 cm(Renzi2013 modified). S. satureioides (Lamiaceae) and C.schoenanthus E Os at different concentrations are tested on 26/04/2018 and 09/05/2018 respectively. In this method 3ml of E O concentration was added in the each box with a dropper on a filter paper (Melisie et al. 2015). A control treatment with 3 ml of distilled water only is adopted. After one honey bee worker was introduced, along with one control box distilled water. This was replicated three times.These boxes were placed on smooth and clean surface at 28 ± 2°C in the dark immediately after treatment application.The mortality was determined after 30 mn, 45mn, 60mn, 90 mn, 120 mn, 150mn and 180mn. Preliminary tests showed that distilled water do not affect the survival and behaviour of honeybees. Determination of mortality Mortality rate of tested honey bees subject to different concentrations of both essential oils is evaluated after 3 hours. Insects are considered as dead when they did not move even when agitating boxes. Mortality is calculated and corrected according to Abbot (1925) formula as follows: Statistical analysis Results analyse is performed using Probit procedure (Finney 1971) todetermine lethal concentration (LC 50 and LC 90 ) using SPSS device. Bees poisoning and locomotion record Photos showing the appearances of dead bees were taken as well as bees’ locomotion activity was registered through video sequences for each essential oil using digital camera (Sony model N°DSC-W150, ISO3200). Bees were observed through video sequences:locomotion activity and contractions were recorded according to a rating scale. Table.1 rating scale adopted for evaluation of locomotion activity and contractions Scale Locomotion activity Contractions 0 No locomotion activity No contraction + Weakly active (less than 1 cm displacement in 10 seconds) weak contraction (less than 2 contractions in 10 seconds) ++ Moderately active (2–3 cm displacement) medium contraction (2–3 contractions in 10 seconds) +++ Very active (more than 3 cm displacement) severe contraction (more than 3 contractions in 10 seconds) Results Mortality of honeybees was assessed after 30, 45, 60, 90, 120, 150 and 180 mn post exposure (Fig. 1 ). The results showed that bees’ mortality in this test increased with increase in concentrations of E O and exposure time. Saccocalyxsatureioides E O is found to be more toxic against Apismellifera L. adult workers at different exposure time periods and concentrations used. With high toxic effect even after minimum period (30 mn) of exposure (Fig. 1 ). Meanwhile higher toxicity is observed after 90 minutes when bees are exposed to different concentrations of E O. The inhalation toxicity of citronella and savory of different concentrations are shown inTable2 below. Table 2 Toxicity by inhalation of essential oil of 5, 10, 20 µl/ml Cymbopogonschoenanthus and Saccocalyxsatureioides on foragers of Apismellifera bees after 3 h of exposure. Values were estimated by Probit Analysis. Essential oils Concentrationµl/ml Mean Mortality LC 50 LC 90 Cymbopogonschoenanthus 5 33,33 7.85 59.85 10 61,90 15 66,66 20 66,66 Saccocalyxsatureioides 5 52,38 6.66 12.43 10 61,90 15 66,66 20 80,94 (P > 0.05, n = 15) The inhalation toxicity test of the essential oils of citronella ( C.schoenanthus ) and savory ( S.satureioides ) showed the last as the most harmful (LC50:6,66µl/ml & LC90: 12.43 µl/ml); whereas the first is the least toxic (LC 50 : 7.85 µl/ml & LC 90 : 59,85 µl/ml) to honey bees workers. The poisoning process recorded by photos showed the tongues of dead bees being extended (Fig. 2 ) after being exposed to C. schoenanthus and S. satureioides inhalation test. Thus, 75% of dead bees exposed to C.schoenanthus have their tongues extended against 83,33%of dead bees exposed to S.satureioides. Disturbance of locomotion and contractions of bees recorded by video sequences registered during exposition to C. schoenanthus and S. satureioides revealed that bees exposed to C. schoenanthus presented lesser contractions and were more active than bees exposed to S. satureioides whom presented more contractions and lesser locomotion activity (Table.3). Table.3 Percentages of locomotion activity, contractions and extended tongues registered during inhalation test. Cymbopogonschoenanthus Saccocalyxsatureioide Scale Locomotion activity Contractions Locomotion activity Contractions 0 14,29 14.29 85,71 0 + 14,29 85.71 14.29 57,14 ++ 28,57 0 0 28,57 +++ 42,85 0 0 14,29 bees with Extended Tongues 75 83.33 DISCUSSION Our study demonstrated C. schoenanthus and S. satureioides E O to be toxic for the honey bee at concentrations over 5 µl/ml by inhalation route in a short time duration 3h. Our reports concerning bees mortality are similar to those of Farooqi et al.(2016); who reported highly toxic effects even after minimum exposure time period of 3 hrs but of Nicotinoids and Spinosad. Thus; the essential oil of C.nardus from Argentina exhibits low toxicity against bees (LC 50 = 11.84 µL/mL) when used to control Varroosis (Giménez-Martínez et al. 2022 ). Compared to C. schoenanthus which is demonstrated to be more toxic to bees in our experiment (LC50 = 6,66µL/mL). This may be attributed to differences in plant species accordingly some researchers reported safety of using some Cymbopogon ’s species with bees such as those of Abramson et al. ( 2006 ) who showed that exposure to the odour of citronella ( C. winterianus ) was not repellent to Africanized honey bees in the northeast of Brazil. Schweitzer et al. ( 2013 ) reported that C.schoenanthus subsp. proximus inflorescences are used for swarms’ attraction in traditional beekeeping in the Burkina Faso. In the same context Santos et al. ( 2018 ) reported that the exposure of A. Mellifera foragers to EO of C. martinii and its major compound (geraniol), in addition to presenting lower toxicity in both routes of exposure, did not present negative effects on the locomotion and orientation of bees under oral exposure. As well as the finding of Sabahi et al. ( 2018 ) who showed that Lemongrass oil ( C. citratus ) have potential for controlling Varroa mites and seem to be relatively safe for larvae and adult honey bees. Extended tongues of bees have been reported as poisoning symptoms as reported by Bradbear ( 2009 ); many of the bees killed by poison will have their tongue extended.Thus; some chemical pesticides caused a locomotor deficit in honeybees (Charreton et al. 2015).Meanwhile; the walking activity of A. mellifera was negatively affected by eucalyptus EO, as well as neem seed kernel oil, which also showed a repellent effect towards honeybee foragers (Xavier et al. 2015 ). Also, oregano EO applied at sublethal doses had negative impact on the distance traveled, the movement speed and the number of stops by A. mellifera (da Silva et al. 2020 ). Our study revealed the toxicity of the essential oils of citronella ( Cymbopogonschoenanthus ) and savory ( Saccocalyxsatureioides ) through inhalation test and indicates the necessity for their conditioned use during blooming periods of flowers. Thus the present findings suggest that there is need to conduct reliable reviews of different bio-pesticides which are bring used on different field crops to guarantee durable development and supervision of beekeeping for better pollination. In general, our results showed that the exposure to S.satureioides is more harmful than the exposure to C.schoenanthus whom the toxicity effect is lesser. Indeed exposure to S.satureioides and C.schoenanthus essential oils to asses’ bees’ toxicity by inhalation route appears to be insufficient. Thus other routes of exposure especially contact and ingestion routes should be studied. Indeed sub-lethal effects on bee behaviour such as trembling, tumbling and lack of coordination should be the purpose of further studies also on the components responsible of toxicity. These preliminary results will also allow for further research on bio-pesticides based on plant essential oils. The laboratory evaluation of inhalation toxicity of E O is only one measure to check their potential insecticide effect, and mortality of honeybees can totally or moderately differ under field conditions due to different factors thus in situ assessments will provide more data in this domain. Conclusion From these findings, we suggest that use of Saccocalyxsatureioides and Cymbopogonschoenanthus essential oils expected to be insecticides could contribute to future improvement actions andconservation of A. mellifera colonies if used out blooming period of growing or after dusk were honey bees are absent. In addition, our study highlights the potential of these E Os as base bio-insecticides for the management of insect pests since these substances are toxic to insect pests and did not influence the mortality of A. Mellifera if used with precautions mentioned before. In conclusion, our preliminary mortality studies showed that bees survived exposure to citronella and Savory at limited concentrations. When the concentration increased, however, theycause mortality with the highest mortality occurring at concentrations above 7.85 and 6.66 µl/ml. Finally our preliminary results must be followed by confirming studies for more concentration and longer periods of time both in vitro and in vivo conditions. Declarations Authors’ contributions K B, S T and F B redaction of the manuscript and participated in the conduct of the inhalation essay, H F and N S realised all steps concerning vegetal material from collection to EO extraction and preparation of different concentrations. Funding The authors have no financial or proprietary interests in any material discussed in this article. Data Availability All data are available in the manuscript. Supplementary datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Ethics approval and consent to participate All authors certify that they have no afliations with or involvement in any organization or entity with any fnancial interest or non-fnancial interest in the subject matter or materials discussed in this manuscript. Consent for publication All authors consent to participate in publication of these data. Competing interests The authors declare that they have no competing interests. References Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entomol 18:265–267. 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Xavier V M, Message D, Picanço, Chediak M, Santana Junior P A, Ramos R S, Martins J C (2015) Acute Toxicity and Sublethal Effects of Botanical Insecticides to Honey Bees.J. Insect Sci.15(1): 137; DOI: 10.1093/jisesa/iev110. Cite Share Download PDF Status: Published Journal Publication published 10 Sep, 2025 Read the published version in International Journal of Tropical Insect Science → Version 1 posted Editorial decision: Major revisions 03 Sep, 2024 Reviewers agreed at journal 16 Aug, 2024 Reviewers invited by journal 16 Aug, 2024 Editor assigned by journal 12 Jan, 2024 First submitted to journal 11 Jan, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3843753","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":341216996,"identity":"3e33d016-b760-4b77-8445-b1c9775ec3c0","order_by":0,"name":"Khalila BENGOUGA","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-0177-8266","institution":"CRSTRA","correspondingAuthor":true,"prefix":"","firstName":"Khalila","middleName":"","lastName":"BENGOUGA","suffix":""},{"id":341216997,"identity":"e5bb89fc-4fa6-44ae-9c86-d14da7d6b3af","order_by":1,"name":"Farida BETTICHE","email":"","orcid":"","institution":"CRSTRA","correspondingAuthor":false,"prefix":"","firstName":"Farida","middleName":"","lastName":"BETTICHE","suffix":""},{"id":341216998,"identity":"18a4fbf1-ab7c-4296-b9d2-23acbfcc7dce","order_by":2,"name":"Souad TAHARCHAOUCHE","email":"","orcid":"","institution":"CRSTRA","correspondingAuthor":false,"prefix":"","firstName":"Souad","middleName":"","lastName":"TAHARCHAOUCHE","suffix":""},{"id":341216999,"identity":"31c5f1da-b8b9-4575-b158-647eefb7452d","order_by":3,"name":"Haroun FADLAOUI","email":"","orcid":"","institution":"CRSTRA","correspondingAuthor":false,"prefix":"","firstName":"Haroun","middleName":"","lastName":"FADLAOUI","suffix":""},{"id":341217000,"identity":"10ef89e8-2677-4de3-a2ec-e5689612ecb0","order_by":4,"name":"Nora SALEMKOUR","email":"","orcid":"","institution":"CRSTRA","correspondingAuthor":false,"prefix":"","firstName":"Nora","middleName":"","lastName":"SALEMKOUR","suffix":""}],"badges":[],"createdAt":"2024-01-07 22:21:58","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3843753/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3843753/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s42690-025-01613-5","type":"published","date":"2025-09-10T15:56:58+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":64438181,"identity":"8f8c7b4a-18dc-4387-803a-f356025c4147","added_by":"auto","created_at":"2024-09-13 07:38:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":49389,"visible":true,"origin":"","legend":"\u003cp\u003eRate of mortality after exposure to different concentrations of both EO.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3843753/v1/d520f25b270df116517056c9.png"},{"id":64438180,"identity":"6483683b-8b13-44ed-bd14-7e237de6de5e","added_by":"auto","created_at":"2024-09-13 07:38:49","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":320952,"visible":true,"origin":"","legend":"\u003cp\u003eDead bees’ extended tongues \u003cstrong\u003ea\u003c/strong\u003e, \u003cstrong\u003eb:\u003c/strong\u003e\u003cem\u003eC. schoenanthus\u003c/em\u003e5 \u0026amp; 20 µl/ml;\u003cstrong\u003ec\u003c/strong\u003e, \u003cstrong\u003ed:\u003c/strong\u003e\u003cem\u003eS.satureioides\u003c/em\u003e10 \u0026amp; 15 µl/ml.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3843753/v1/7ced872045455847668f3684.png"},{"id":91359148,"identity":"b8e071ee-c877-46b0-bb42-4964ee2572e9","added_by":"auto","created_at":"2025-09-15 16:05:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":982258,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3843753/v1/a6c3d19b-0ccc-425b-9013-dbc5454b3a97.pdf"}],"financialInterests":"","formattedTitle":"The Citronella and Savory Essential Oils Effect on Honeybees (Apis mellifera L.) Mortality","fulltext":[{"header":"Background","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eInsecticides are used on different crops to control their insect-pests. Along with harmfulinsects these insecticides also kill the beneficial insects especially pollinators like honey bees (Husain et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).The tremendous increase in crop yields associated with the \u0026lsquo;green\u0026rsquo; revolution has been possible in part by the discovery and utilization of chemicals for pest control. However, concerns over the potential impact of pesticides on human health and the environment has led to the introduction of new pesticide registration procedures, such as the Food Quality Protection Act in the United States. These new regulations have reduced the number of synthetic pesticides available in agriculture. Therefore, the current paradigm of relying almost exclusively on chemicals for pest control may need to be reconsidered. New pesticides, including natural product-based pesticides are being discovered and developed to replace the compounds lost due to the new registration requirements (Dayan et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). For that recourse to natural derived insect control agents such as essential oils is proposed as apromising alternative to protect beneficial insects. As their effectiveness on pathogen agents was proved especially of some plants essential oils such: basil, colocynth, jasmine, geranium, senna, chamomileas (Sharaby et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2020\u003c/span\u003e),aloe, ginger, garlic, and hot pepper (Siam and Othman \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), neem (Gopal et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTo prove the antibactirial and fungicidal activities of the essential oils of \u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e(Poaceae) and\u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e (Lamiaceae); different studies were maintained in Algeria. High efficacy against E. coli, an average efficacyagainst Staphylococcusaureus and a limited efficacy against Pseudomonasaerogenosof \u003cem\u003eS.satureioides\u003c/em\u003e E O was reported byLahrech (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). It was reported also as having a broad spectrum of activity as it inhibits both the growth of bacteria that moulds and yeasts. (Ariech and Hendel \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The results obtained by Sari et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) indicate that the E O of Saccocalyx possesses an intermediate anti-leishmania activity in vitro. Fungicidal activity of \u003cem\u003eC.schoenanthus\u003c/em\u003e E O is also reported by Amraoui(2014).As well Guerrah\u0026amp;Segueni (2015)demonstrated methanolic extract of \u003cem\u003eC.schoenanthus\u003c/em\u003e, antimicrobial activity against E.coli.\u003c/p\u003e \u003cp\u003eMeanwhile insecticidal activity of \u003cem\u003eC. schoenanthus\u003c/em\u003e emulsified oil can validly be used as alternative in management of \u003cem\u003eP. Xylostella\u003c/em\u003e larvaeon cabbages (Laba2012). Results on the Effect of insecticide formulation based essential oil of \u003cem\u003eC.schoenanthus\u003c/em\u003e (L.)aplicated on cotton in Togo showed a significant reduction of cotton pests \u003cem\u003eAphis gossypii\u003c/em\u003eGlover (Homoptera :Aphididae) population and an improvement in the production in accordance with the increase of the concentration of \u003cem\u003eC. schoenanthus\u003c/em\u003e essential oil (Bokobana et al.2016).For now in Algeria Aous et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) studied antioxidant, antimicrobial and insecticidal properties of this plant specieds. Meanwhile the effects of both plants extracted oils to honeybees have been briefly researchedthus,Fuselli et al. (2010) have tested citronella grass (\u003cem\u003eC.nardus\u003c/em\u003e) and palmarosa (\u003cem\u003eC.martinii\u003c/em\u003e) from Gramineae to potentially control \u003cem\u003ePaenibacillus\u003c/em\u003e larvae, the causal agent of American foulbrood disease (AFB).\u003c/p\u003e \u003cp\u003eIn order to promote local aromatic plants in the fight against crop pests and reduce the use of synthetic chemical pesticides and the risks associated with them, the effectiveness of an insecticide formulation based essential oil was elaborated by a project proposed by CRSTRA aiming to control \u003cem\u003eTutaabsoluta\u003c/em\u003e on tomatoes and \u003cem\u003eAphis gossipy\u003c/em\u003e on melon; thus two essential oils of two plants \u003cem\u003eC.schoenanthus\u003c/em\u003e (Poaceae) and \u003cem\u003eS.satureioides\u003c/em\u003e (Lamiaceae) were exploited.\u003c/p\u003e \u003cp\u003eBut as all other fight methods, the use of pesticide plants presents some favours as well as some limits (Yarou et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Our subject is whether this option does not have any effect on beneficial insects especially on pollinators such as \u003cem\u003eApismellifera\u003c/em\u003e.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv\u003e\n \u003cp\u003eThe aim of the present study was to evaluate toxic inhalation of \u003cem\u003eC. schoenanthus\u003c/em\u003e and \u003cem\u003eS. satureioides\u003c/em\u003e to bees\u0026rsquo; workers.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec3\"\u003e\n \u003ch2\u003eVegetal material\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003ePlants of both species \u003cem\u003eC. schoenanthus\u003c/em\u003e and \u003cem\u003eS. satureioides\u003c/em\u003e were collected from Djelfa and Bousaadasteppic regions of Algeria during April 2016 and 2017. The plants were dried up for a period of one month in aired obscure place. Every two days plants were verified and turned to avoid mould.\u003c/p\u003e\n \u003cp\u003e100 g dried leaves of both plants were subjected to hydro-distillation by a Clevenger device of 1000 ml flask for 4 hours at 100\u0026deg;C.Extracted essential oil was diluted using distilled water.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\"\u003e\n \u003ch2\u003eBees\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003eAdult honeybee workers of \u003cem\u003eApismellifera\u003c/em\u003e L. were collected from the bees\u0026rsquo; hives from Bio-Ressources station of CRSTRA located at El-Outaya. The hives at the time the bees were collected were free of diseases. No hive treatments to control diseases were conducted before collection of bees. Honeybees were collected from the flight board at the hive entrance at morning time of the day of the experiment, (OEPP 2010) and putted into a plastic container and were brought to laboratory.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\"\u003e\n \u003ch2\u003eInhalation Test\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003eTreatments used in the bioassays are the EO extracted of both plants where solutions of 5, 10, 15 and 20 \u0026micro;l/ml are prepared in distilled water on the same day just before the test. The surface residual method in glass jars (Radwan and Taha 2012) with some modifications is adopted for testing contact toxicity of different concentrations of E Os to honeybees (\u003cem\u003eA. mellifera\u003c/em\u003e).Within plasticboxes(Detzeland Wink 1993) of 15x20x30 cm(Renzi2013 modified).\u003cem\u003eS. satureioides\u003c/em\u003e (Lamiaceae) and \u003cem\u003eC.schoenanthus\u003c/em\u003e E Os at different concentrations are tested on 26/04/2018 and 09/05/2018 respectively. In this method 3ml of E O concentration was added in the each box with a dropper on a filter paper (Melisie et al. 2015). A control treatment with 3 ml of distilled water only is adopted. After one honey bee worker was introduced, along with one control box distilled water. This was replicated three times.These boxes were placed on smooth and clean surface at 28\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C in the dark immediately after treatment application.The mortality was determined after 30 mn, 45mn, 60mn, 90 mn, 120 mn, 150mn and 180mn.\u003c/p\u003e\n \u003cp\u003ePreliminary tests showed that distilled water do not affect the survival and behaviour of honeybees.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\"\u003e\n \u003ch2\u003eDetermination of mortality\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003eMortality rate of tested honey bees subject to different concentrations of both essential oils is evaluated after 3 hours. Insects are considered as dead when they did not move even when agitating boxes. Mortality is calculated and corrected according to Abbot (1925) formula as follows:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\"\u003e\n \u003ch2\u003eStatistical analysis\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003eResults analyse is performed using Probit procedure (Finney 1971) todetermine lethal concentration (LC\u003csub\u003e50\u003c/sub\u003e and LC\u003csub\u003e90\u003c/sub\u003e) using SPSS device.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\"\u003e\n \u003ch2\u003eBees poisoning and locomotion record\u003c/h2\u003e\n \u003cdiv\u003e\n \u003cp\u003ePhotos showing the appearances of dead bees were taken as well as bees\u0026rsquo; locomotion activity was registered through video sequences for each essential oil using digital camera (Sony model N\u0026deg;DSC-W150, ISO3200). Bees were observed through video sequences:locomotion activity and contractions were recorded according to a rating scale.\u003c/p\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003eTable.1\u0026nbsp;\u003c/strong\u003erating scale adopted for evaluation of locomotion activity and contractions\u003c/p\u003e\n \u003cdiv\u003e\n \u003ctable id=\"Taba\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eScale\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLocomotion activity\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eContractions\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo locomotion activity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo contraction\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWeakly active (less than 1 cm displacement in 10 seconds)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eweak contraction (less than 2 contractions in 10 seconds)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eModerately active (2\u0026ndash;3 cm displacement)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003emedium contraction (2\u0026ndash;3 contractions in 10 seconds)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVery active (more than 3 cm displacement)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esevere contraction (more than 3 contractions in 10 seconds)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eMortality of honeybees was assessed after 30, 45, 60, 90, 120, 150 and 180 mn post exposure (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe results showed that bees\u0026rsquo; mortality in this test increased with increase in concentrations of E O and exposure time. \u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e E O is found to be more toxic against \u003cem\u003eApismellifera\u003c/em\u003e L. adult workers at different exposure time periods and concentrations used. With high toxic effect even after minimum period (30 mn) of exposure (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Meanwhile higher toxicity is observed after 90 minutes when bees are exposed to different concentrations of E O.\u003c/p\u003e \u003cp\u003eThe inhalation toxicity of citronella and savory of different concentrations are shown inTable2 below.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eToxicity by inhalation of essential oil of 5, 10, 20 \u0026micro;l/ml \u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e and\u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e on foragers of \u003cem\u003eApismellifera\u003c/em\u003e bees after 3 h of exposure. Values were estimated by Probit Analysis.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEssential oils\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eConcentration\u0026micro;l/ml\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean Mortality\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLC\u003csub\u003e50\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eLC\u003csub\u003e90\u003c/sub\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e33,33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e7.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e59.85\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e61,90\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66,66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66,66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e52,38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e6.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e12.43\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e61,90\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e66,66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e80,94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e(P\u0026thinsp;\u0026gt;\u0026thinsp;0.05, n\u0026thinsp;=\u0026thinsp;15)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eThe inhalation toxicity test of the essential oils of citronella (\u003cem\u003eC.schoenanthus\u003c/em\u003e) and savory (\u003cem\u003eS.satureioides\u003c/em\u003e) showed the last as the most harmful (LC50:6,66\u0026micro;l/ml \u0026amp; LC90: 12.43 \u0026micro;l/ml); whereas the first is the least toxic (LC\u003csub\u003e50\u003c/sub\u003e: 7.85 \u0026micro;l/ml \u0026amp; LC\u003csub\u003e90\u003c/sub\u003e: 59,85 \u0026micro;l/ml) to honey bees workers.\u003c/p\u003e \u003cp\u003eThe poisoning process recorded by photos showed the tongues of dead bees being extended (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) after being exposed to \u003cem\u003eC. schoenanthus\u003c/em\u003eand \u003cem\u003eS. satureioides\u003c/em\u003einhalation test. Thus, 75% of dead bees exposed to\u003cem\u003eC.schoenanthus\u003c/em\u003ehave their tongues extended against 83,33%of dead bees exposed to \u003cem\u003eS.satureioides.\u003c/em\u003e\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eDisturbance of locomotion and contractions of bees recorded by video sequences registered during exposition to \u003cem\u003eC. schoenanthus\u003c/em\u003e and \u003cem\u003eS. satureioides\u003c/em\u003erevealed that bees exposed to \u003cem\u003eC. schoenanthus\u003c/em\u003e presented lesser contractions and were more active than bees exposed to \u003cem\u003eS. satureioides\u003c/em\u003e whom presented more contractions and lesser locomotion activity (Table.3).\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eTable.3\u003c/b\u003e Percentages of locomotion activity, contractions and extended tongues registered during inhalation test.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabb\" border=\"1\"\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e\u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e\u003cem\u003eSaccocalyxsatureioide\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eScale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLocomotion activity\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eContractions\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLocomotion activity\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eContractions\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14,29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e14.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e85,71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e14,29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e85.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e14.29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57,14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e28,57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e28,57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e+++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e42,85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14,29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ebees with Extended Tongues\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e83.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eOur study demonstrated \u003cem\u003eC. schoenanthus\u003c/em\u003eand \u003cem\u003eS. satureioides\u003c/em\u003eE O to be toxic for the honey bee at concentrations over 5 \u0026micro;l/ml by inhalation route in a short time duration 3h. Our reports concerning bees mortality are similar to those of Farooqi et al.(2016); who reported highly toxic effects even after minimum exposure time period of 3 hrs but of Nicotinoids and Spinosad.\u003c/p\u003e \u003cp\u003eThus; the essential oil of \u003cem\u003eC.nardus\u003c/em\u003e from Argentina exhibits low toxicity against bees (LC\u003csub\u003e50\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;11.84 \u0026micro;L/mL) when used to control Varroosis (Gim\u0026eacute;nez-Mart\u0026iacute;nez et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Compared to \u003cem\u003eC. schoenanthus\u003c/em\u003e which is demonstrated to be more toxic to bees in our experiment (LC50\u0026thinsp;=\u0026thinsp;6,66\u0026micro;L/mL). This may be attributed to differences in plant species accordingly some researchers reported safety of using some \u003cem\u003eCymbopogon\u003c/em\u003e\u0026rsquo;s species with bees such as those of Abramson et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) who showed that exposure to the odour of citronella (\u003cem\u003eC. winterianus\u003c/em\u003e) was not repellent to Africanized honey bees in the northeast of Brazil. Schweitzer et al. (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) reported that \u003cem\u003eC.schoenanthus\u003c/em\u003e subsp. \u003cem\u003eproximus\u003c/em\u003e inflorescences are used for swarms\u0026rsquo; attraction in traditional beekeeping in the Burkina Faso. In the same context Santos et al. (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported that the exposure of \u003cem\u003eA. Mellifera\u003c/em\u003e foragers to EO of \u003cem\u003eC. martinii\u003c/em\u003e and its major compound (geraniol), in addition to presenting lower toxicity in both routes of exposure, did not present negative effects on the locomotion and orientation of bees under oral exposure. As well as the finding of Sabahi et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) who showed that Lemongrass oil (\u003cem\u003eC. citratus\u003c/em\u003e) have potential for controlling Varroa mites and seem to be relatively safe for larvae and adult honey bees.\u003c/p\u003e \u003cp\u003eExtended tongues of bees have been reported as poisoning symptoms as reported by Bradbear (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2009\u003c/span\u003e); many of the bees killed by poison will have their tongue extended.Thus; some chemical pesticides caused a locomotor deficit in honeybees (Charreton et al. 2015).Meanwhile; the walking activity of \u003cem\u003eA. mellifera\u003c/em\u003e was negatively affected by eucalyptus EO, as well as neem seed kernel oil, which also showed a repellent effect towards honeybee foragers (Xavier et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Also, oregano EO applied at sublethal doses had negative impact on the distance traveled, the movement speed and the number of stops by \u003cem\u003eA. mellifera\u003c/em\u003e (da Silva et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur study revealed the toxicity of the essential oils of citronella (\u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e) and savory (\u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e) through inhalation test and indicates the necessity for their conditioned use during blooming periods of flowers. Thus the present findings suggest that there is need to conduct reliable reviews of different bio-pesticides which are bring used on different field crops to guarantee durable development and supervision of beekeeping for better pollination.\u003c/p\u003e \u003cp\u003eIn general, our results showed that the exposure to \u003cem\u003eS.satureioides\u003c/em\u003e is more harmful than the exposure to \u003cem\u003eC.schoenanthus\u003c/em\u003e whom the toxicity effect is lesser.\u003c/p\u003e \u003cp\u003eIndeed exposure to \u003cem\u003eS.satureioides\u003c/em\u003e and \u003cem\u003eC.schoenanthus\u003c/em\u003e essential oils to asses\u0026rsquo; bees\u0026rsquo; toxicity by inhalation route appears to be insufficient. Thus other routes of exposure especially contact and ingestion routes should be studied. Indeed sub-lethal effects on bee behaviour such as trembling, tumbling and lack of coordination should be the purpose of further studies also on the components responsible of toxicity. These preliminary results will also allow for further research on bio-pesticides based on plant essential oils.\u003c/p\u003e \u003cp\u003eThe laboratory evaluation of inhalation toxicity of E O is only one measure to check their potential insecticide effect, and mortality of honeybees can totally or moderately differ under field conditions due to different factors thus in situ assessments will provide more data in this domain.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eFrom these findings, we suggest that use of \u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e and \u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e essential oils expected to be insecticides could contribute to future improvement actions andconservation of \u003cem\u003eA. mellifera\u003c/em\u003e colonies if used out blooming period of growing or after dusk were honey bees are absent. In addition, our study highlights the potential of these E Os as base bio-insecticides for the management of insect pests since these substances are toxic to insect pests and did not influence the mortality of \u003cem\u003eA. Mellifera\u003c/em\u003e if used with precautions mentioned before.\u003c/p\u003e \u003cp\u003eIn conclusion, our preliminary mortality studies showed that bees survived exposure to citronella and Savory at limited concentrations. When the concentration increased, however, theycause mortality with the highest mortality occurring at concentrations above 7.85 and 6.66 \u0026micro;l/ml. Finally our preliminary results must be followed by confirming studies for more concentration and longer periods of time both in vitro and in vivo conditions.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eK B, S T and F B redaction of the manuscript and participated in the conduct of \u0026nbsp;the inhalation essay, H F and N S realised all steps concerning vegetal material from collection to EO extraction and preparation of different concentrations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no financial or proprietary interests in any material discussed in this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data are available in the manuscript. Supplementary datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors certify that they have no afliations with or involvement in any organization or entity with any fnancial interest or non-fnancial interest in the subject matter or materials discussed in this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors consent to participate in publication of these data.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAbbott WS (1925) A method of computing the effectiveness of an insecticide. 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Environ. 21(4): 288-304.\u003c/li\u003e\n \u003cli\u003eSharaby A M F, Gesraha M A, Fallatah S A B (2020)Botanical extracts against the potato tuber moth, Phthorimaeaoperculella (Zeller 1873) (Lepidoptera: Gelechiidae), during storage conditions. Egypt.\u0026nbsp;J.\u0026nbsp;Biol.\u0026nbsp;Pest Control./doi.org/10.1186/s41938-020-00294-1.\u003c/li\u003e\n \u003cli\u003eXavier V M, Message D, Pican\u0026ccedil;o, Chediak M, Santana Junior P A, Ramos R S, Martins J C (2015) Acute Toxicity and Sublethal Effects of Botanical Insecticides to Honey Bees.J. Insect Sci.15(1): 137; DOI: 10.1093/jisesa/iev110. \u0026nbsp;\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Essential oils, Toxicity, Honey bee","lastPublishedDoi":"10.21203/rs.3.rs-3843753/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3843753/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u0026nbsp;Recently a trend to use small concentrations of the essential oils of (\u003cem\u003eCymbopogonschoenanthus\u003c/em\u003e) or (\u003cem\u003eSaccocalyxsatureioides\u003c/em\u003e) to control pests has extended. They are often used to suppress biotic enemies such as fungus, moulds, bacteria and insects. What is not known is whether these oils also influence beneficial beings such as honeybees. In preliminary experiments foragers’ bees were exposed by inhalation to different concentrations of \u003cem\u003eC. schoenanthus\u003c/em\u003e) or (\u003cem\u003eS. satureioides\u003c/em\u003e essential oils. Results showed that bees’ mortality in inhalation test increased with increase in concentrations of essential oils and exposure time. Both essential oils showed LC\u003csub\u003e50\u003c/sub\u003e greater than 5 µl/ml.Meanwhile 205 µl/ml concentrations were more detrimental. Disturbance of locomotion and contractions of bees recorded by video sequences registered revealed that bees exposed to C. schoenanthus presented lesser contractions and were more active than bees exposed to S. satureioides whom presented more contractions and lesser locomotion activity. Our research provides further evidence supporting the use of conditioning techniques to evaluate the use of essential oils as optimal substances to be considered as alternative controls of pests assuming honey bee safety.\u003c/p\u003e","manuscriptTitle":"The Citronella and Savory Essential Oils Effect on Honeybees (Apis mellifera L.) Mortality","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-09-13 07:38:44","doi":"10.21203/rs.3.rs-3843753/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revisions","date":"2024-09-03T11:18:18+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-08-16T19:56:54+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-16T15:55:54+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-01-12T07:00:02+00:00","index":"","fulltext":""},{"type":"submitted","content":"International Journal of Tropical Insect Science","date":"2024-01-11T08:30:27+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"3bc5a83a-17fb-4fb2-8946-d57e98197ee4","owner":[],"postedDate":"September 13th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-09-15T16:03:42+00:00","versionOfRecord":{"articleIdentity":"rs-3843753","link":"https://doi.org/10.1007/s42690-025-01613-5","journal":{"identity":"international-journal-of-tropical-insect-science","isVorOnly":false,"title":"International Journal of Tropical Insect Science"},"publishedOn":"2025-09-10 15:56:58","publishedOnDateReadable":"September 10th, 2025"},"versionCreatedAt":"2024-09-13 07:38:44","video":"","vorDoi":"10.1007/s42690-025-01613-5","vorDoiUrl":"https://doi.org/10.1007/s42690-025-01613-5","workflowStages":[]},"version":"v1","identity":"rs-3843753","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3843753","identity":"rs-3843753","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}