Acta agriculturae Slovenica, 119/1, 1–13, Ljubljana 2023 doi:10.14720/aas.2023.119.1.2707 Original research article / izvirni znanstveni članek Bags impregnated with garlic (Allium sativum L.) and parsley (Petrose- linum crispum (Mill.) Fuss) essential oils as a new biopesticide tool for Trogoderma granarium Everts, 1898 pest control Alzahraa Abdelaty ELMADAWY 1, Ahmed Fayez OMAR 2, 3, Tamer ISMAIL 2 Received May 28, 2022; accepted January 11, 2023. Delo je prispelo 28. maja 2022, sprejeto 11. januarja 2023 1 Plant Protection Research Institute, Stored Product Pests Department, Agricultural Research Center, Ministry of Agriculture, Giza, Egypt 2 Pesticides Chemistry and Toxicology Department, Faculty of Agriculture, Kafrelsheikh University, Kafrelsheikh, Egypt 3 Corresponding author, e-mail: ahmed.foz.fayez9@gmail.com Bags impregnated with garlic (Allium sativum L.) and pars- ley (Petroselinum crispum (Mill.) Fuss) essential oils as a new biopesticide tool for Trogoderma granarium Everts, 1898 pest control Abstract: Stored product pests can cause significant damages and great economic problems in stored commodities and grain cereal. Using synthetic pesticides in the storage pest control has adverse effects on human health. In our study, the toxicity of garlic (Allium sativum) and parsley (Petroselinum crispum) essential oils (EOs) impregnating with three types of bags were assessed against Trogoderma granarium Everts, 1898 adults after different exposure intervals. GC-MS analysis of the investigated EOs revealed that the major components of parsley and garlic were 1, 3, 8-p-menthatriene (23.34 %) and di-allyl disulfide (27.9 %), with (1.40 %) alpha-terpinene and (1 %) of di-alyl tetra-sulfide as minor components respectively. Addi- tionally, comparison the toxicity among the treated bags was assessed based on the LC50 values and a persistence efficiency of the tested EOs was carried out by the LC90 values for each bag type. In all bag types, garlic and parsley had mortality by 100 % for clothes, and 80 % for both plastic and paper bags after 7 days of exposure, respectively. After two and five days of garlic treatment, plastic bags were the most effective, but after seven days of exposure paper bags was more effective than the other two types. Finally, cloth bags treated with EOs were the most effective packaging for insect control, indicating that this ap- proach could be considered as an additional tool to the concept of stored product management.  Key words: essential oils; stored grain; bags; Trogoderma granarium Vrečke, impregnirane z eteričnimi olji česna (Allium sativum L.) in peteršilja (Petroselinum crispum (Mill.) Fuss) kot novo biopesticidno orodje za nadzorovanje indijskega žitnika (Tro- goderma granarium Everts, 1898) Izvleček: Skladiščni škodljivci lahko povzročijo znatno škodo in velike ekonomske probleme v skladiščih žit .Uporaba sintetičnih pesticidov pri nadzoru skladiščnih škodljivcev ima škodljive učinke na zdravje ljudi. V raziskavi so bili preučeva- ni toksični učinki eteričnih olj česna in peteršilja na odrasle osebke indijskega žitnika s tremi tipi vrečk (plastične, papirna- te in iz blaga), impregniranimi z eteričnimi olji obeh rastlin v različnih časovnih izpostavitvah. GC-MS analiza preučevanih eteričnih olj je pokazala, da sta bili njihovi glavni sestavini 1, 3, 8-p-mentatrien (23, 34 %) in dialil disulfid (27,9 %), z (1,40 %) alfa-terpinenom in (1 %) dialil tetrasulfidom v manjšem deležu. Dodatno je bila ocenjena primerjava strupenosti impregnira- nih vrečk na osnovi LC50 vrednosti. Trajnost učinka eteričnih olj je bila preiskušena na osnovi LC90 vrednosti za vsak tip vreč- ke. Primerjalno je impregniranost vrečk z eteričnimi olji česna in peteršilja povzročila 100 % smrtnost pri vrečkah iz blaga in 80 % smrtnost pri plastičnih in papirnatih vrečkah po sedem- dnevni izpostavitvi. Po izpostavitvi za dva in pet dni v vrečkah impregniranih s česnom so bile najbolj učinkovite plastične vrečke, po sedmih dneh izpostavitve so bile najbolj učinkovite papirnate vrečke. Zaključimo lahko, da so vrečke iz blaga, im- pregnirane z eteričnimi olji najbolj učinkovite za uravnavanje škodljivih žuželk in bi jih lahko uporabili kot dodatno orodje pri ohranjanju shranjenih pridelkov. Ključne besede: eterična olja; shranjena zrna; vrečke; Trogoderma granarium Acta agriculturae Slovenica, 119/1 – 20232 A. A. ELMADAWY et al. 1 INTRODUCTION Stored commodities are vulnerable to insect attack. Major stored grain cereal crops can be attacked by more than 600 species of beetles, moths, and other insect pests and mites, causing a great economic problem either qual- itatively or quantitatively deteriorating (Rajendra and Sriranjini, 2008). According to estimates, more than one- third of food products are lost during post-harvest stor- age due to pest infestation (Tripathi et al., 2009). Trogo- derma granarium Everts (Coleoptera: Dermestidae), the khapra beetle, is one of the most important storage pests, which has economic importance under severe phytosan- itary worries (Myers and Hagstrum 2012; EPPO 2013; Athanassiou et al., 2019). This species has been classified among the most harmful and intrusive outsider species on the planet (Lowe et al., 2000). The potential expansion of T. granarium is due to its ability to attack numerous stored cereals or related products and non-grain com- modities. It feeds on 96 commodities and infests very dry food in a very dry environment with 2 % relative humid- ity (r. h.). In addition to its ability to shuttle from one place to another according to worldwide exchange and temperature tolerance, which ranged between (21 and 40 °C) with an optimum at 35 °C (Degri and Zainab 2013; Athanassiou et al., 2016; Kavallieratos et al., 2019). Their larvae may fall under selective diapause for a long and show resistance to insecticides (Edde et al., 2012; Myers and Hagstrum, 2012; Athanassiou et al., 2015). Due to the widespread overuse of synthetic pesticides in manag- ing insect pests, including those attacking stored prod- ucts, and the development of resistances, efforts to re- duce environmental pollution represent a major issue of concern for environmental and health issues (Desneux et al., 2007). Due to expansion instances of khapra beetle re- sistance against traditional insecticides such as phos- phine, malathion and pyrethroids have more featured its monetary importance and raised another errand for the researchers to seek a new way to control it (Myers and Hagstrum, 2012; Honey et al., 2017; Khalique et al., 2018). In this way, the European Union supports a con- siderable decrease in the utilization of insecticides, by looking for less harmful substances under the Integrated Pest Management (IPM) models (Hillocks, 2012; Lucchi and Benelli 2018). During the last years, botanicals such as plant powder and essential oils have been evaluated as promising alternatives for the control of a huge range of pests on different-stored commodities (Athanassiou et al., 2014; Bohinc et al., 2020). Plant essential oils are com- plex natural mixtures of volatile organic compounds re- sulting as secondary metabolites in plants, constituted by terpenes, terpenoids, and phenol-derived aromatic com- ponents and, aliphatic components (Bakkali et al., 2008). They have been reported by many studies that essential oils have toxic effects such, as a repellent, antifeedant, antifungal, antibacterial, reproduction, and inhibitory development effects against varied insect pests (Ebadol- lahi and Jalali Sendi 2015; Tu et al., 2018; Hu et al., 2019). Packing is one of the most common methodologies for the development of bioactive preservation tools. It is confirmed that packaging offers a basic point in food quality conservation and a definitive defense against in- sect pests. Active packaging is considered a promising in- novation for food packaging. It consists of incorporating into the packaging material active compounds useful for food protection. Earlier studies mentioned that packages are one of the possible ways to protect stored products during storage until they reach consumers (Campbell et al., 2004). Furthermore, naturally derived compounds such as essential oils and plant extracts consolidated in active packaging can be used in place of commonly used chemical preservatives by consumers who prefer natural preservatives (Bazargani-Gilani et al., 2015). However, insect pests can infest packaged products in many ways (Mullen et al., 2012; Costa, 2014; Stejskal et al., 2017). Endeavors ought to be made not just towards the plan- ning of successful frameworks, which might hinder the food quality rot, but also towards the improvement of insect-proof packages, ready to stand up against insect penetration and/or to repel their presence from the food package’s environment. Recently, many essential oils have been employed as promising natural preservatives in packaging. A lot of exhaustive research has developed numerous packaging material types that can offer different levels of protection for stored products at a low cost for either individuals or companies (Stejskal et al., 2017). Treated packages protect the stored grain commodities by blocking insect infestations resulting from the storage environmental conditions (Paudyal et al., 2017a). Similarly, a covering containing citronella oil is applied to the containerboard for insect repellence (Wong et al., 2005). Consequently, insecticides or other protective materials can be impreg- nated into the synthesis of the storage bags or treated straightforwardly on the packaging material (Scheff et al., 2016; Kavallieratos et al., 2017a, b; Paudyal et al., 2017a, b; Scheff et al., 2017; Scheff and Arthur 2018). Active agents consolidated in packing materials could be released through evaporation of the headspace or can migrate from the package to the product by diffusion through the packaging material prolonging the specific activity and reducing the migration to the preserved ma- terial. For instance, antimicrobial cellulose packaging was achieved through the laccase-interceded joining of phenolic compounds (Elegir et al., 2008). Therefore, the Acta agriculturae Slovenica, 119/1 – 2023 3 Bags impregnated with garlic ... and parsley ... essential oils as a new biopesticide tool for Trogoderma granarium ... pest control objectives of this study were to evaluate the immediate and delayed mortality of T. granarium adults exposed to three types of available and low-cost storage bags made of plastic, paper, and clothes material impregnated with two natural essential oils garlic and parsley addition to evaluating their storage long activity. 2 MATERIALS AND METHODS 2.1 INSECT Trogoderma granarium Everts, 1898 (Coleoptera: Dermestidae) used in the bioassays were obtained from stock colonies maintained in the laboratory of Stored Product Insects of the Sakha Agricultural Research Sta- tion, Agriculture Research Center (ARC) Egypt. Adults of T. granarium < 24 h old and larvae having 2–4 mm long (Athanassiou et al., 2016) were used in the experi- ments, which were cultured on wheat at 35 °C, 65 % rela- tive humidity (RH), and continuous darkness. 2.2 ESSENTIAL OIL Garlic (Allium sativum L.) and parsley (Petrose- linum crispum [P. Mill.] Nyman ex A.W. Hill) EOs are widely available in Egypt, and they were supplied by Hashem Brothers Company for Essential Oils and Aro- matic Products (Kafr-Elsohby, Kalyoubeya, Egypt). 2.3 GC/MS ANALYSIS OF THE ESSENTIAL OIL The chemical composition of the essential oils was analyzed by gas chromatography-mass spectrometry (GC/MS) using the model (HP5890- USA) provided by an HP column (60 m × 0.25 mm, 0.25 μm film thick- ness) (HP-5 ms). The initial temperature was 60 °C and the maximum temperature was 250 °C for 65.3 min. The injector temperature was 240 °C. Relative percent- age amounts were calculated from the peaks’ total area by apparatus software. The compounds were identified by matching the mass spectra data with those held in a computer library (Wiley 275. L), according to Swigar and Silverstein (1981) and Adams (1995). All analysis steps were carried out at the laboratory of Hashem Brothers Company, Egypt. 2.4 STORAGE BAGS Three different types of bags were used in the exper- iments: polypropylene (BOPP) bags that are utilized for containing and moving products like food sources, pro- duce, powders, ice, magazines, chemicals, and waste it is a common form of packaging. Kraft paper (KP) which are some of the most popular forms of packaging for con- sumers and finally clothes bags, which are considered a popular alternative to plastic and paper shopping bags, because cloth bags do not cause the environmental harm as plastic bags. 2.5 BIOASSAYS 2.5.1 Contact The standard solution of each tested essential oil (EO) was obtained by diluting 1 ml of crude oil in 100 ml acetone then, four concentrations of each essential oil, (1.25, 2.5, 5, and 7.5 mg ml-1) of garlic, & (7.5, 10, 12.5, and 15 mg ml-1) of parsley were prepared from the stand- ard solution of garlic and parsley then impregnated into tested bags. Each concentration was sprayed into the test- ed bags with 1 ml as a fine mist that contained the appro- priate concentration of each oil. Bags were splashed with a brush on both sides. After treatment with each EOs, the brush was carefully cleaned with acetone and subse- quently, the following treatment was applied to the other type. An additional series of bags were prepared without any treatment and served as controls. The sprayed bags were left to become dry for 24 h (h) at 30 °C and 65 % r. h. Then, in each type of bag, 20 g of wheat was placed, and ten adults of Trogoderma granarium were put in each bag and then closed with stapler staples. Each concentra- tion was replicated three times for each type and placed in incubators set at 30 C and 65  % r. h. Accumulative mortality counts were recorded at 2, 5, and 7 days after treatment corrected by Abbott’s formula (1925). 2.6 LONG-TERM EXPERIMENT Long–term experiment was conducted to evaluate the persistence and long-active effectiveness of various concentrations of testing essential oils loaded with testes bags, this experiment was carried out as follows. After calculating the LC90 value of testing essential oil for each type of bag individually, 9 bags of each type were treated with the 2 days calculated LC90 value of garlic and parsley and stored treated until further investigation. After one month of storage three bags of each treated type (paper, plastic, and cloth) were infected with 10 adults of T. gra- narium insect and another three untreated bags served as a control, mortality was observed after 2 days. This ex- Acta agriculturae Slovenica, 119/1 – 20234 A. A. ELMADAWY et al. periment was repeated after three months as mentioned above and recorded mortality rate. 2.7 STATISTICAL ANALYSIS The results were analyzed by one-way analysis of variance ANOVA followed by the Least Significant Dif- ference test for mean separation. p values of ≤ 0. 05 were considered significant. The experiments were performed in triplicate, the data presented are the mean ± SE. The lethal concentration for 50  % mortality (LC50) was de- termined by log-probit analysis (Finney, 1971), and the data were analyzed by determining chi-square values and degrees of freedom. The analysis of data was performed with SPSS program version 24.0 for Windows (SPSS Inc., IBM Corp.) 3 RESULTS AND DISCUSSION 3.1 CHEMICAL COMPOSITION TESTED ESSEN- TIAL OILS The major components of testing essential oils are summarized in Tables 1 & 2. A total of 33 components were identified for parsley essential oil according to their retention indices in Table 1, the major constitu- ents representing 84.63 % of the total component were trimethyl bicylo (13.01  %), beta pinene (8.28  %), beta myrcene (3.93  %), beta phellandrene (3.81  %),1,3,8-p- menthatriene (23.34  %), benzodioxole (11.67  %), apiol (12.72 %) and benzofuran (7.87 %). Whereas the minor constituents were chavicol (2.51 %), benzene-methyl (2.06 %), and alpha-terpinene (1.40 %). Where in Table 2 a total of 42 components were identified for garlic es- sential oil according to their retention indices, the major constituents representing 94.4 % of the total component were dimethyl disulfide (1.4 %), diallyl sulfide (9.5 %), al- lyl methyl disulfide (8.3 %), dimethyl trisulfide (2.9 %), diallyl disulfide ( 27.9 %), allyl (Z)-1 propenyl disulfide (2.2  %), allyl (E)-1-propenyl disulfide (3.7  %), allyl methyl trisulfide (17.7  %), 4-methyl-1,2,3-trithiolane (1.2 %), 2-vinyl-4H-1,3-dithiine (1.8 %), diallyl trisulfide (16.8 %) and diallyl tetrasulfide (1 %). Essential oils from various plant species could rep- resent an alternative way to existing synthetic pesticide agents, either by direct application or by loading on a car- rier. In our study chemical composition of parsley (EO) P. crispum agrees with earlier studies from different re- gions all over the world (Zhang et al., 2006; Soher et al., 2014) with some differences in component percentage due to climate change and environmental factors. Also, the chemical composition of garlic (EO) A. sativum is in agreement with those (Satyal et al., 2017; Mossa et al., 2018), who reported that garlic oil from different topo- graphical areas has shown subjective similitudes, yet quantitative contrasts in the groupings of the organosul- fur compound. 3.2 TOXICITY BIOASSAY In the current study A. sativum and P. crispum, es- sential oils exhibited strong insecticidal activity against T. granarium adults with a significant difference between treated concentrations along the test period and within bag types. Where the results demonstrate that A. sati- vum oil showed high efficiency over P. crispum oil against khapra beetle with all tested bags. Mortality percentage for garlic increased as a function of time exposure, where the highest concentration of garlic recorded 100 % mor- tality in clothes bags, 86.66 % for plastic bags, and 96.66 % for paper bags after 7 d of exposure (Fig 1c). On the other hand, parsley recorded mortality by 80 % for plastic Compounds Percent Composition % Molecular formula Retention time (min) Trimethyl bicylo 13.01 C10H16 6.20 Beta pienen 8.28 C10H16 7.52 Beta myrcene 3.93 C10H16 7.97 Beta phellandrene 3.81 C10H16 9.31 1,3,8-p-menthatriene 23.34 C10H14 12.85 Benzodioxole 11.67 C7H6 O2 30.05 Apiol 12.72 C12H14 O4 36.02 Benzofuran 7.87 C8H6 O 12.08 Table 1: Main components of parsley (Petroselinum crispum) essential oil analyzed by gas chromatography-mass spectrometry (GC-MS) Acta agriculturae Slovenica, 119/1 – 2023 5 Bags impregnated with garlic ... and parsley ... essential oils as a new biopesticide tool for Trogoderma granarium ... pest control Compounds Percent Composition % Molecular formula Retention time(min) Dimethyl disulfide 1.4 C2H6S2 12.32 Diallyl sulfide 9.5 C6H10S2 14.25 Allyl methyl disulfide 8.3 C4H8S2 15.26 Dimethyl trisulfide 2.9 C2H6S3 16.13 Diallyl disulfide 27.9 C6H10S2 18 Allyl (Z)-1-propenyl disulfide 2.2 C4H8S2 18.21 Allyl (E)-1-propenyl disulfide 3.7 C6H10S2 18.33 Allyl methyl trisulfide 17.7 C4H8S3 18.96 4-Methyl-1,2,3-trithiolane 1.2 C3H6S3 19.21 2-Vinyl-4H-1,3-dithiine 1.8 C6H8S2 20.23 Diallyl trisulfide 16.8 C6H10S3 21.68 Diallyl tetrasulfide 1 C6H10S4 25.66 Table 2: Main components of garlic (Allium sativum) essential oil, analyzed by gas chromatography-mass spectrometry (GC-MS) Tested oil Time (d) LC50 Value (mg kg-1) Confidence Interval 95 % Slope value Chi-Square (X2)Lower Upper Garlic 2 days 19.26 a 24.06 15.39 1.165 0.035 5 days 9.86 d 7.88 12.33 0.599 0.238 7 days 1.240 f 0.99 1.55 0.107 0.874 parsley 2 days 17.98 b 15.00 22.48 1.575 0.598 5 days 12.01 c 7.508 15.02 1.022 0.483 7 days 3.65 e 2.92 4.56 0.257 0.325 Table 3: Toxicity assessment of garlic and parsley essential oils against Trogoderma granarium adults after different exposure peri- ods in plastic bags Values in the row followed by the same letters are not significantly different (p > 0.05) according to ANOVA and Duncan multiple-comparison tests Tested oil Time (d) LC50 Value (mg kg-1) Confidence Interval 95 % Slope value Chi-Square (X2)Lower Upper Garlic 2 days 6.31 d 5.05 7.89 0.504 0.600 5 days 5.21 e 4.17 8.08 0.641 2.085 7 days 4.33 f 3.46 7.07 0.717 0.087 parsley 2 days 18.89 a 14.94 23.61 1.869 0.347 5 days 16.14 a 12.91 20.18 1.22 0.086 7 days 8.34 c 3.54 10.08 1.187 0.505 Table 4: Toxicity assessment of garlic and parsley essential oils against Trogoderma granarium adults after different exposure periods in paper bags Values in the row followed by the same letters are not significantly different (p > 0.05) according to ANOVA and Duncan multiple-comparison tests Acta agriculturae Slovenica, 119/1 – 20236 A. A. ELMADAWY et al. migant toxicity than phosphine against Callosobruchus maculatus (Fabricius, 1775). In addition, Kavallieratos et al. (2020) mentioned that eight essential oils could be considered grain protectants to manage adults and lar- vae of T. granarium. Janaki et al. (2018) showed that T. granarium adults were highly susceptible to the Cyperus rotundus L. EO when applied to filter paper at 0.04 μl cm- 2, causing 94 % repellence after 2 h of exposure. A comparison between tested bags to illustrate the most powerful type of bags results in Fig. 3, showed that paper and clothes bags impregnated with garlic oil was the most effective one after two and five days post- treatment, but with increasing the investigation period to seven days clothes bags showed high effectiveness than the other two types. However, in parsley treatment, Fig. 4 clothes bags were the most effective one than the other two bag types during the tested period. Coating is the most common methodology for the advancement of bioactive packing, our bioassay was es- tablished for the evaluation of insect control by oil-im- pregnated packaging through three types of bags. Bags are a trustworthy method of storage, due to the expansion of the assurance they can give against insect invasions is of high significance (Paudyal et al., 2017a). Previous studies have demonstrated the efficiency of loading dif- ferent toxic agents against stored product insects. Wong et al. (2005) indicated that commercial citronella could reduce beetle infestation, initially by approximately 50 % when applied at 0.2 g m-2 of carton board in addition to its repellent effect for 16 weeks when applied directly as a coating into carton board in ethanol solution. Our results are in line with Kavallieratos et al. (2017a) who documented that deltamethrin impregnated into ZeroFly storage bags was very effective against larger grain borer Prostephanus truncatus (Horn, 1878) by 94.4 % after 5 d of exposure, the rice weevil Sitophilus oryzae (L., 1763) by 100 % after 1 d of exposure, and by 96.7 % on the ware- bags, 80 % for paper bags, and 73.33 % for clothes bags at the highest concentration after 7 d of exposure as shown in Fig. 2 c. In Table 3, A. sativum essential oil had the lowest LC50 values by 19.26, 9.86, and 1.24 mg kg -1 after two, five, and seven days respectively, opposite to 17.98, 12.01, and 3.65 mg kg-1 for P. crispum for plastic bags treatment. The same was in both paper and clothes bags (Tables 4 & 5), where LC50 values were 6.31, 5.21, and 4.33 mg kg -1 for A. sativum compared to 18.89, 16.14, and 8.34 mg kg-1 for P. crispum after two, five and seven days in paper bags treat- ment, while clothes bags recorded LC50 values by 6.13, 2.24, and 1.211 mg kg-1 for A. sativum oil compared to 6.63, 3.32, and 1.47 mg kg-1 for P. crispum after the same investigation period. In this framework, our findings indicate that es- sential oils impregnated into different types of bags can contribute to managing one of the stored-product insect species with economic importance Trogoderma grana- rium. The EO of A. sativum was the most effective among the tested EOs against T. granarium, causing a higher mortality rate than P. crispum oil with all types of test- ing bags. Botanicals incorporate a wide scope of prom- ising compounds used for developing novel, effective, and ecologically economical pesticides (Stevenson et al., 2017; Pavela et al., 2019a). Extracts and oils of garlic have already been advertised as pest control products. Yang et al. (2009) found that garlic essential oil loaded on coated polyethylene glycol (PEG) in the size of the na- noparticles could effectively control Tribolium castaneum (Herbst) insect. In addition, George et al. (2010) found that the garlic oil was highly toxic to the red poultry mite, Dermanyssus gallinae (De Geer, 1778). On the other side, the insecticidal activity of parsley has been demonstrated in some studies on arthropods (Bortolucci et al., 2015; Mansour et al., 2015). Whereat, Massango et al. (2016) reported that the essential oil of parsley showed low fu- Tested oil Time (d) LC50 Value (mg kg-1) Confidence Interval 95 % Slope value Chi-Square (X2)Lower Upper Garlic 2 days 6.13 b 4.68 9.65 1.013 0.214 5 days 2.24 d 0.144 3.47 0.438 1.95 7 days 1.211 f 0.76 2.18 0.366 2.685 parsley 2 days 6.63 a 4.52 8.29 0.732 0.828 5 days 3.32 c 2.66 4.15 0.189 0.055 7 days 1.47 e 1.17 3.59 0.162 0.183 Table 5: Toxicity assessment of garlic and parsley essential oils against Trogoderma granarium adults after different exposure periods in clothes bags Values in the row followed by the same letters are not significantly different (p > 0.05) according to ANOVA and Duncan multiple-comparison tests Acta agriculturae Slovenica, 119/1 – 2023 7 Bags impregnated with garlic ... and parsley ... essential oils as a new biopesticide tool for Trogoderma granarium ... pest control Fig 1: Mortality percentage of Trogoderma granarium adults in bags treated with garlic (Allium sativum) essential oil after different exposure periods (a, b and c) house beetle, Trogoderma variabile (Ballion, 1878) after 5 d of exposure. In the same trend, Kavallieratos et al. (2017b) found that treated bags with chlorfenapyr and pirimiphos-methyl were effective to kill all adults of P. trunactus and the lesser grain borer, Rhyzopertha domini- ca (F., 1792) (Coleoptera: Bostrychidae) after 3 d of expo- sure. The same was reported by Paudyal et al. (2017a) on S. oryzae and the red flour beetle, T. castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae) adults. Our results showed that all bag types had a mortality efficiency against tested insects, whereas Kavallieratos and Bouk- ouvala (2018) in their studies reported that three tested types of bags showed equal mortality levels of T. grana- rium. In addition, Herrera et al. (2021) reported that the Acta agriculturae Slovenica, 119/1 – 20238 A. A. ELMADAWY et al. Fig 2: Mortality percentage of Trogoderma granarium adults in bags treated with parsley (Petroselinum crispum) essential oil after different exposure periods (a, b and c) Acta agriculturae Slovenica, 119/1 – 2023 9 Fig. 3: Comparison between bags types treated with garlic oil (Allium sativum) according to LC50 value Fig. 4: Comparison between bags types treated with parsley oil (Petroselinum crispum) according to LC50 value Bags impregnated with garlic ... and parsley ... essential oils as a new biopesticide tool for Trogoderma granarium ... pest control Acta agriculturae Slovenica, 119/1 – 202310 A. A. ELMADAWY et al. insecticidal effect of the biopesticide of the silo bag with the added essential oil of Mentha x piperita L. released from the silo bag against R. dominica (F.) showed 100 % mortality during the time examined therefore silo bag could be used to control different pests in stored grain. Also, Vendl et al. (2021) mentioned that the treatment of different pieces of food packaging, cardboard spacer, and pallet with bergamot oil had critical repellent activ- ity against Sitophilus granarius and Tribolium confusum Jacquelin du Val, 1863. The storage experiment summarized in Table 6 showed that all tested bags had lasting toxic efficiency against adult T. granarium along the storage period that extended for three months. Where after one month of storage mortality percentage ranged from 90 to 70 % with the superiorly of paper bags over clothes and plas- tic bags for both tested essential oils respectively. On the other side, garlic oil was significantly more effective than parsley with all bag types after 3 months of storage, where the mortality rate was 20, 40, and 33.3 % for paper, plas- tic, and clothes bags respectively compared to 16.6, 30, and 26.66 in parsley. In the long-term experiment, results indicate that LC90 concentration of garlic and parsley essential oils applied to tested bag types could produce protection to stored grains against T. granarium infection for a while lasting for three months. This technique offers the pos- sibility of developing a new approach with many advan- tages. Amalraj et al. (2020) reported that black pepper and ginger essential oils consolidated in chitosan- (CS), gum arabic– (GA), and polyethylene glycol (PEG) showed high antimicrobial activity against a wide range of bacteria such as Bacillus cereus Frankland & Frankland 1887, Staphylococcus aureus Rosenbach 1884, Escherichia coli Migula 1895, and Salmonella typhimurium Lignières 1900, as a promising option in food packaging and wound dressing materials. Also, Mapossa et al. (2021) reported that studies searched for controlled-release for- mulations could be effective against mosquito vectors of malaria parasites. Moreover, plant powder from Xylopia aethiopica (Dunal) A. Rich. can serve as a carrier when mixed with its essential oil giving longer-term protection for grain against Callosobruchus maculatus insect due to lipids or other components present in the fruit powder may adsorb the terpenes, slowing down their release in the flask as reported by Habiba et al. (2010). Fig 5: Storage efficiency of garlic oil (Allium sativum) and parsley (Petroselinum crispum) oil impregenated in different bags types after one and three months Bags Mortality (%) after 1 month of storage Mortality (%) after 3 months of storage Garlic Parsley Garlic Parsley Paper 90 b 86.66 b 20 bc 16.66 bc Plastic 73.33 b 70 b 40 b 30 b Cloth 76.66 b 76.66 b 33.33 b 26.66 bc Control 0 a 0 a 0 a 0 a Table 6: Mortality percentage of Trogoderma granarium adults exposed to bags treated with LC90 value of garlic and parsley essen- tial oils after different storage times Values in the row followed by the same letters are not significantly different (p > 0.05) according to ANOVA and Duncan multiple-comparison tests Acta agriculturae Slovenica, 119/1 – 2023 11 Bags impregnated with garlic ... and parsley ... essential oils as a new biopesticide tool for Trogoderma granarium ... pest control 4 CONCLUSION Controlling T. granarium adults could be attainable by impregnating both garlic and parsley essential oil in three types of storage bags polypropylene (BOPP), kraft paper (KP), and clothes. Garlic oil had a greater mortal- ity effect on T. granarium than parsley oil. The BOPP and clothes bags showed higher mortality than (KP) bags with both garlic and parsley oil after 7 days of exposure. It was revealed that the treatment of clothes bags is the most effective approach for protecting grain against stor- age pests, which might have significant ramifications for the utilization under industrial conditions. For long- term storage results indicated that all tested bags had lasting toxicity effectiveness extended for 3 months vary- ing from one type to another, whereas paper bags (KP) was the most lasting one over the other two types. 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