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Khapra beetle, Trogoderma granarium Everts, 1898 (Coleoptera: Dermestidae) is one of the most notorious pests of stored grains. This study was aimed to evaluate the efficacy of ozone against 2nd and 5th instars and adults of T. granarium in stored wheat kernels under laboratory conditions. Four ozone concentrations of 300, 600, 900, and 1200 parts per million by volume (ppmv) were used. The results revealed that the LC values were 249.76 ppmv for adult, 446.75 ppmv for 5th instar, and 275.30 ppmv for 2nd instar of T. granarium after 2 h exposure to ozone. No adults emerged when the ozone concentration was 1200 ppmv, while emergence was high in controls (91.20%) after a 1-h exposure period. The highest ozone concentration of 1200 ppmv resulted in strong grain protection against adult and 2nd and 5th instars, with a 0.63%, 0.73%, and 1.16% loss in grain weight, respectively. Chemical analysis of treated wheat kernels at a concentration of 1200 ppmv ozone showed no significant differences (p > 0.05) in fat, moisture, ash, carbohydrate, fiber, and protein content of ozone-treated wheat kernels compared to untreated grain. Thus, ozone can be effectively used to control T. granarium and provides sufficient protection for stored wheat. Keywords Khapra beetle · Ozone · Mortality · Adult emergence · Weight loss Introduction internal-feeding insect pests of stored grain. It feeds on a wide range of cereals such as wheat, rice, oats, barley, and Cereals are the main source of calories in many countries. rye (Kavallieratos et al. 2019; Papanikolaou et al. 2019). It Wheat (Triticum aestivum L.) is the most important cereal is ranked as one of the 100 worst invasive species worldwide crop because, it provides about 20 percent of total food (Usman et al. 2018). This beetle is widespread in regions calories for the human race (Rathore 2001). Stored grains with high temperatures and low humidity, and is especially require continuous protection to avoid deterioration of qual- prevalent in some areas of the Middle East, South America, ity and weight especially caused by insects (Padin et al. Africa, Asia, and Europe (Eliopoulos 2013; Ghanem and 2013). Insect infestation leads to 10–20% cereal loss sup- Shamma 2007; Harris 2009). The most destructive stage is ply in worldwide, human health risks (malnutrition) beside the larvae, which feed inside the grain, leading to quantita- the loss of millions of dollars annually (Nagpal and Kumar tive and qualitative losses during storage (Athanassiou et al. 2012; Pedigo and Rice 2014). 2019; Kavallieratos et al. 2017; Omar et al. 2012). Infected The khapra beetle, Trogoderma granarium (Everts) grain attracts external-feeding insect pests and fungi, conse- (Coleoptera: Dermestidae), is one of the most destructive quently leading to the deterioration of the grain and loses its market value, reduces the nutritional value of the grain, and becomes unfit for human use (Arain et al. 2006; Madkour * M. A. Mahmoud et al. 2012). MahmoudAli.email@example.com Control of insects in stored products depends primarily Plant Protection Department, Faculty of Agriculture, Al- on the application of insecticides, where injudicious use of Azhar University, Assiut, Egypt insecticides and fumigants cause serious problems such as Plant Protection Department, Faculty of Agriculture, toxic residues in food grains (Wasala et al. 2015). Addition- Al-Azhar University, Cairo, Egypt ally, there are worries about the environment and human Center of Plasma Technology, Al-Azhar University, Cairo, health, rising pesticide costs, the emergence of insect pests Egypt Vol.:(0123456789) 1 3 Applied Entomology and Zoology with resistance, unfavourable effects on organisms other than Materials and methods targets, and insect pests (Islam and Ahmed 2016; Meyer- Baron et al. 2015; Pourya et al. 2018). In recent years, a Insect rearing lot of study has concentrated on alternatives to insecticides for controlling stored materials, such as heating, radiation, The beetles used in this study were obtained from the biological control using bacteria and fungi as well as plant Plant Protection Institute, Ministry of Agriculture, Giza, extracts (Ibrahim and Al-Nasser 2009; Mohammed et al. Egypt. Adults of T. granarium were cultured in an incu- 2019). Ozone is an effective, as well as promising, alter - bator at a constant temperature of 30 °C ± 2 °C, 65 ± 5% native to chemical fumigants as a viable green insecticide relative humidity and photoperiod 12:12 h (light:dark) in against a range of stored-grain insects, which prevents the the laboratory at Plant Protection Department, Faculty need to store and dispose of hazardous chemicals; there is no of Agriculture, Al-Azhar University, Cairo Governorate. need for aeration to remove the gas after application (Isikber Before being utilized as a substrate for insect rearing, the and Athanassiou 2015). Ozone has been used in the food experiment's wheat variety (Sakha 95) was deinfested in industry for more than 100 years, and it was first used as an oven at 60 °C for one hour. Stock culture was set up the food preservative of frozen meat in 1910 (Carletti et al. by introducing one hundred adults pairs and confined for 2013). However, growing emphasis has been placed on the 7 days for oviposition into glass jars of one liter capacity positive influence of ozone application in agriculture, such each containing 500 gm wheat kernels and covered with as the utilization of ozone in food processing for inactivation muslin held in place by rubber bands. The culture was of pathogens, reduction of storage pests and degradation of reared to obtain homogeneous stock of 2nd and 5th instars various mycotoxins (Bonjour et al. 2011). Application of and adults in separate jars. The homogeneous stock was ozone in stored-products is effective in controlling insects maintained for further experiments. and microorganisms such as fungi and bacteria, and degrad- ing pesticide residues (Tiwari et al. 2010). Ozone was found to be an effective environment friendly alternative against T. Production of ozone gas granarium, since it is a highly oxidative toxic gas, which is a potent insect killer and degrades rapidly to oxygen (Mahroof Ozone gas was produced using an ozone generator cylin- and Amoah 2018). It is efficient and economical, and does drical dielectric barrier discharge (DBD) at the Laboratory not leave a residue on stored processed products (Mendez of the Center of Plasma Technology, Al-Azhar University, et al. 2003; Tiwari et al. 2010). Ozone has a different efficacy Nasr City, Cairo, Egypt. against insects inside or outside the grain, mainly because In the gas generation process, oxygen was used as the grains can increase the decomposition of ozone (Işikber input, passing through a DBD reactor. The discharge is and Öztekin 2009). Many researchers have applied ozone produced by applying a discharge voltage between two to various stored commodities and it is effective against a coaxial electrodes, with a glass dielectric between them wide range of stored insect pests such as Sitotroga cerealella and a free space where the oxygen flows. In this free space, (Olivier), Tribolium castaneum (Herbst), Sitophilus oryzae a filament discharge is produced, generating electrons with (L.), Callosobruchus maculatus (F.), Callosobruchus chin- enough energy to break down the oxygen molecules, form- ensis (L.), Ephestia cautella (Walk.), Plodia interpunctella ing ozone (O ). The DBD cell was fed with oxygen gas. (Hübner), T. granarium and Rhyzopertha dominica (F.) (Abd The concentration of the generated ozone was controlled El-Ghaffar et al. 2016, 2017; Amoah and Mahroof 2019; by the discharge current, and the oxygen gas flow rate Gad et al. 2021a; Ghazawy et al. 2021; Işikber and Öztekin was adjusted to 0.5 l/min. Ozone was applied directly into 2009; Mishra et al. 2019; Pandiselvam et al. 2019; Sabeat jars containing the T. granarium larvae and adults with and Sabr 2015; Subramanyam et al. 2017; Xinyi et al. 2019). the grains. The ozone gas is constantly renewed inside Therefore, preliminary laboratory studies were carried the exposure fumigation chamber. This renewal and the out to evaluate the efficacy of ozone gas on the 2nd and continuous flow of ozone gas during the exposure period 5th instars and adults of T. granarium, as one of the major of insects works to avoid the half-life time of the ozone, economic stored grain pests in Egypt. The effects of various which ranges from 20 to 30 min. This continuous flow ozone concentrations on insect mortality and adult emer- makes the ozone gas pass constantly through the fumiga- gence, as well as the harm caused to wheat kernels, were tion chamber, which contains the insects inside and con- studied. stantly renewed. The ozone concentration inside the jars 1 3 Applied Entomology and Zoology was measured using an ozone analyzer (Model H1-AFX- oxygen flow rate 0.5 (l/min) at different ozone concentra- Instrumentation, USA). The input voltage of the AC test tions of 300, 600, 900, and 1200 ppmv is 3.4, 4.0, 5.6, and set (specifically, a variable high voltage transformer) was 6.8 kV, respectively. 220 V at 50 Hz. A voltage transformer was connected to two outer and inner electrodes, separated by a gap. Volt- Power measurement method ages used to generate ozone were controlled through a transformer control box (Variac Variable AC Power Trans- The power is analyzed following the original work of Man- former Regulator) as shown in Fig. 1. ley (1943), who has utilized voltage–charge Lissajous fig- ures to characterize the average consumed power through the discharge (Ballinger et al. 2005; Biganzoli et al. 2014; Characterization of electrical properties Janeco et al. 2011; Manley 1943). The charge–voltage char- acteristic plot is revealed in Fig. 3. The two values effective The current–voltage oscillogram of the coaxial DBD reactor discharge capacitance is indicated by obtaining two distinct system, recorded at atmospheric pressure and room tempera- slopes of the Q–V plot. The reactor (coaxial dielectric bar- ture, is presented in Fig. 2. The filamentary modes discharge rier discharge) power formula was shown in equations by current in DBD when the applied voltage exceeds the break- Manley (1943), where the total power (Pel) is related to the down voltage. operating frequency (ƒ) and the peak voltageV . From max In Fig. 2, waveforms of the applied voltage to coaxial this so-called Lissajous Figure, the minimum external volt- DBD reactor and associated current measured for ozone at age V at which the ignition occurs, the electric energy min Fig. 1 Schematic diagram of the ozone generation and fumiga- tion setup 1 3 Applied Entomology and Zoology 5 30 5 60 4kV Ozoneconcentration=300 ppmv Ozone concentration=600ppmv 3.4kV 25 4 4 20 40 3 3 2 2 10 20 1 1 0 0 0 0 -5 -1 -1 -10 -20 -2 -2 -15 -3 -3 -20 -40 -4 -4 -25 -5 -30 -5 -60 -0.05-0.04-0.03-0.02-0.010.000.010.020.030.040.05 -0.05-0.04-0.03-0.02-0.010.000.010.020.030.040.05 Time (ms) Time (ms) 8 60 10 40 Ozoneconcentration=1200ppmv Ozoneconcentration=900 ppmv 9 5.6kV 6.8kV 5 6 5 20 4 30 1 1 0 0 0 0 -1 -5 -1 -10 -2 -10 -2 -3 -20 -15 -4 -3 -5 -20 -4 -30 -6 -25 -5 -7 -40 -30 -6 -8 -50 -35 -9 -7 -10 -40 -8 -60 -0.05-0.04-0.03-0.02-0.010.000.010.020.030.040.05 -0.05-0.04-0.03-0.02-0.010.000.010.020.030.040.05 Time (ms) Time (ms) Fig. 2 Waveforms of the applied voltage to reactor and associated current measured for ozone at oxygen flow rate 0.5 (l/min) Fig. 3 Lissajous diagrams 300 ppmv measured for ozone at flow rate 900 ppmv 1.00 600 ppmv of 0.1 (l/min) Qmax 1.00 0.75 0.50 Q 900 ppmv 0.25 max 0.00 V 0.75 -10 -8 -6 -4 -2 0 246 810 1200 ppmv Voltage (kV) -0.25 min -0.50 C =dQ/dV -0.75 0.50 C=dQ/dt -1.00 0.25 0.00 -7 -6 -5 -4 -3 -2 -1 01234567 Voltage (kV) -0.25 -0.50 P =0.12 Watt el P =0.17 Watt -0.75 el P =0.47 Watt el -1.00 P =1.12 Watt el 1 3 Appliedvoltage (kV) Applied voltage (kV) Charge (C µ ) Dischargecurrent (mA) Discharge current (mA) Appliedvoltage (kV) Applied voltage (kV) Charge (C) Dischargecurrent (mA) Discharge current (mA) Applied Entomology and Zoology consumed per voltage cycle E and the electric power (P ) Eec ff t of ozone gas on chemical constituents el el can be estimated by the following relations (Ballinger et al. of wheat kernels 2005): E = 2(V Q − Q V ) ≡ area of (Q−V) diagram Chemical analysis was conducted using a near-infrared el max max 0 spectroscopy (model DA1650 e FOSS Corporation, Den- el mark) to determine the effect of ozone on the ash, carbo- P = = fE (1) el el hydrate, fat, fiber, moisture, and protein levels in a 50 g sample of infested wheat kernels untreated and treated In this study, the consumed power was determined to with ozone at a concentration of 1200 ppmv for 1 h (Taha be 0.12 W at applied voltage of 3.4 kV, 0.17 W at 4.0 kV, et al. 2016). Carbohydrate content was calculated accord- 0.47 W at 5.6 kV and 6.8 kv at 1.12 watts. Treatments were ing to AOAC (2010): % carbohydrate content = [100 − (% applied in case of ozone concentration where ƒ is frequency protein + % moisture + % ash + % lipids + % fiber)]. of 50 Hz, as shown in Fig. 2 and 3. Eec ff t of ozone gas application on the khapra beetle Statistical analysis Each muslin cloth bag containing 50 g of wheat kernels Data were subjected to analysis of variance (ANOVA) received 16 T. granarium larvae in their 2nd or 5th instars or using the Statistical Analysis System at a 5% signifi- eight pairs of adults (1–3 days old) separately. Rubber bands cance level. The mean differences were separated using were used to tightly shut the bag. The bags were exposed the Least Significant Difference (LSD) and are shown as to direct ozone application in a fumigation chamber. Four means ± SE. Probit analysis was conducted to reckon L C ozone concentrations were used (300, 600, 900, and 1200 and confidence limits using SPSS 16.0 software (Chicago, ppmv) with exposure of 1 h or 2 h. Three replicates were IL, USA). made for each treatment (sixteen larvae or eight pairs of adults per replicate). After exposure, the treated groups were transferred (insects and wheat kernels) carefully into glass Results jars (250 ml capacity each) that were covered with a muslin cloth, and secured with rubber bands. The control treatment Eec ff t of ozone gas on mortality of larvae and adults was the same but without ozone exposure. Seven days after of T. granarium exposure to treatments, the mortality of T. granarium lar- vae and adults was counted, and all adults were removed. The probit estimates from the mortality response of 2nd and Then, all jars were retained under the same conditions. The 5th instars and adults of T. granarium exposed to different mortality percentages were corrected using Abbott's formula ozone concentrations are summarized in Table 1. The results (1925). Data on pupal and adult formation were recorded revealed that the L C values were 266.08 ppmv for adult, until the emergence of F1 adults. The percent reduction 280.29 ppmv for 2nd instar and 691.0 ppmv for 5th instar of of pupal and adult emergence was calculated according to T. granarium after 1 h exposure to ozone while, the effect of Silassie and Getu (2009) using the formula: ozone after 2 h exposure with L C values 275.30, 446.75 Reduction rate (%) = Cn − Tn/Cn × 100;where Cn is the and 249.76 ppmv was recorded for the 2nd and 5th instars number of newly emerged insects in the control group, and and adults of T. granarium, respectively. The results showed Tn is the number of newly emerged insects in a treatment a negative relationship between exposure time and L C val- group. ues. The results showed that, regardless of exposure time, Sixty days after treatment, the number of F1 adults that the toxicity of ozone was adult ˃ 2nd instar ˃ 5th instar. The emerged was counted and recorded. Also, the percent of slope values ranged from 1.71 to 2.44, indicating a homoge- adult emergence was calculated using the following formula nous effect across the insects in their susceptibility to ozone. (Howe 1971): Adult emergence (%) = Number of adult emerged/Number of pupae × 100. Eec ff t of ozone gas on adult emergence The percent weight loss of the grains was estimated after 60 days following the method given by Jackai and Asante The data in Table 2 shows the effect of ozone gas concentra- (2003): tions on T. granarium adult emergence after 1 h of treatment. Weight loss (%) = IW – FW/IW × 100; where IW is the The results show that the mean number of adults emerged initial weight and FW is the final weight. from untreated grains was 37.33. Ozone was most effective at 900 and 1200 ppmv, since no adult beetles emerged, while 1 3 Applied Entomology and Zoology Table 1 Toxicity of ozone gas against T. granarium 2nd and 5th instars and adults after exposure periods of 1 and 2 h Exposure Stage Mean ± SE time (hrs) a b c 2 d e f LC (ppmv) 95% Confidence limits (ppmv) Slope Intercept (χ ) df p Lower Upper 1 2nd instar 280.29 ± 199.08 164.46 ± 163.98 445.42 ± 234.38 2.26 ± 0.36 − 6.32 ± 1.04 0.2 2 0.9 5th instar 691.0 ± 319.30 383.95 ± 313.87 26,713.24 ± 25,291.5 1.71 ± 0.01 − 5.11 ± 0.11 0.34 2 0.84 Adult 266.08 ± 45.68 10.53 ± 10.53 448.74 ± 51.18 1.76 ± 0.09 − 4.24 ± 0.27 1.82 2 0.45 2 2nd instar 275.30 ± 39.34 67.48 ± 40.55 413.59 ± 36.47 2.44 ± 0.13 − 5.96 ± 0.47 1.31 2 0.65 5th instar 446.75 ± 28.42 205.73 ± 36.99 621.03 ± 38.88 2.36 ± 0.14 − 6.25 ± 0.39 2.55 2 0.3 Adult 249.76 ± 42.07 34.05 ± 33.36 407.35 ± 37.72 2.07 ± 0.16 − 4.96 ± 0.52 1.18 2 0.58 The concentration causing 50% mortality Slope of the concentration-mortality regression line Intercept of the regression line Chi square value Freedom degree Probability value Table 2 Effect of ozone gas on adult emergence of T. granarium after Eec ff t of ozone gas on pupation and adult an exposure of 1 h emergence of 2nd and 5th instars Concen- Mean ± SE tration Table 3 indicates the effect of ozone gas on pupation and No. of adult emer- Adult emergence Reduction (%) (ppmv) adult emergence of 2nd and 5th instars after a 1-h exposure. gence (%) For the 2nd instar, the pupation reduction of T. granarium a a 0 37.33 ± 0.88 91.20 ± 1.82 – were 58.49 and 96.29%, while adult emergence reduction b ab 300 3.66 ± 0.33 65.71 ± 7.19 90.12 ± 1.09 were 78.63 and 100%, at 300 and 1200 ppmv, respectively. c b 600 1.66 ± 0.33 55.55 ± 5.53 95.53 ± 0.88 For the 5th instar, the pupation reduction were 81.64 and cd c 900 0.33 ± 0.33 16.66 ± 16.68 99.07 ± 0.92 100%, while adult emergence reduction were 93.63 and d c 1200 0.00 ± 0.00 0.00 ± 0.00 100.00 ± 0.00 100%, at 300 and 1200 ppmv, respectively. F 1170 18.9 Eec ff t of ozone gas on weight loss Means in the same column, followed by the same letter are not sig- nificantly different at 0.05 level of probability (df = 4) Table 4 shows mean percentage of wheat kernels weight loss when treated with ozone gas at four concentrations com- the mean number of adult emergence was 3.66 adults at 300 pared with untreated. The mean percentage weight loss of ppmv. The percent reduction of adult emergence was 99.07% the ozone-untreated grains were 5.06%, 5.06%, and 3.56% and 100% at 900 and 1200 ppmv, respectively. for the 2nd and 5th instars and adults, respectively. The Table 3 Effect of ozone gas on Concentration Mean reduction (%) ± SE pupation and adult emergence (ppmv) of 2nd and 5th instars of T. 2nd instar 5th instar granarium after a 1-h exposure Pupation Adult emergence Pupation Adult emergence period b b c a 300 58.49 ± 8.17 78.63 ± 6.30 81.64 ± 4.74 93.63 ± 3.19 ab ab bc a 600 77.52 ± 7.11 93.33 ± 6.67 87.45 ± 2.93 96.96 ± 3.33 a a ab a 900 93.51 ± 3.34 96.66 ± 3.33 96.96 ± 3.03 100.00 ± 0.00 a a a a 1200 96.29 ± 3.71 100.00 ± 0.00 100.00 ± 0.00 100.00 ± 0.00 F 8.52 3.72 7.14 1.76 Means in the same column, followed by the same letter are not significantly different at 0.05 level of prob- ability (df = 3) 1 3 Applied Entomology and Zoology Table 4 Effect of ozone gas on loss in grain weight caused by T. gra - Discussion narium 2nd and 5th instars and adults after a 1-h exposure period The study indicated that ozone is effective against T. gra - Concentration Mean weight loss (%) ± SE (ppmv) narium, and the toxicology data show that mortality hap- 2nd instar 5th instar Adult pens quickly and gets worse with continued ozone expo- a a a 0 5.06 ± 0.24 5.06 ± 0.24 3.56 ± 0.24 sure. These observations are in agreement with Abo-El-Saad b b b 300 3.63 ± 0.27 3.46 ± 0.57 2.82 ± 0.03 et al. (2011) who demonstrated that ozonation at 2.0 ppmv b bc c 600 3.06 ± 0.54 2.86 ± 0.06 1.90 ± 0.05 caused 83% and 27% mortality against E. cautella adults b c d 900 2.7 ± 0.36 2.20 ± 0.37 1.30 ± 0.34 and larvae after 12 h respectively. Bonjour et al. (2011) c d e 1200 0.73 ± 0.12 1.16 ± 0.06 0.63 ± 0.08 reported that 50 ppmv ozone for 4 d caused 100% mortality F 21.1 19.6 40.5 of immature P. interpunctella. According to McDonough et al. (2011), adults of P. interpunctella died completely after Means in the same column, followed by the same letter are not sig- being exposed to 500 ppmv of ozone for 60 min. According nificantly different at 0.05 level of probability (df = 4) to Hansen et al. (2012), the successful implementation of ozone technology necessitates an enough concentration of ozone gas concentration of 300 ppmv resulted in grain pro- ozone for a suitable period of time, which changes depend- tection against 2nd and 5th instars and adults with a 3.63%, ing on the environmental factors. Hussain (2014) reported 3.46%, and 2.82% loss in grain weight, respectively. Moreo- that mortality percentages gradually increased by increas- ver, the highest ozone concentration of 1200 ppmv resulted ing the exposure period to ozone gas, when E. cautella in strong wheat kernels protection against adult and 2nd and larvae were exposed to 80 ppmv ozone, the mortality rate 5th instars, with a 0.63%, 0.73%, and 1.16% loss in grain was 38.92% for a 1-h exposure period. Subramanyam et al. weight, respectively. Generally, all treatments with ozone (2014) examined the toxicity of ozone at two concentra- gas to control T. granarium significantly reduced the loss in tions (0.43 and 0.86 g/m ) against adults of R. dominica grain weight compared to untreated. and demonstrated that the insect was highly susceptible to ozone treatment; the toxicity of ozone after 5 d was higher Eec ff t of ozone gas on wheat kernels quality than that after 1 d of treatment. According to Jemni et al. (2015), exposing Ectomyelois ceratoniae (Zeller) larvae Figure 4 shows the percentages of fat, moisture, ash, car- to 170.7 ppmv of ozone resulted in 82% mortality in just bohydrate, fiber, and protein in ozone-treated and untreated 80 min. According to Sabeat and Sabr (2015), the appli- wheat kernels. The fat percentage in ozone-treated grains cation of ozone to suppress different T. granarium stages were 1.85%, 1.44%, and 1.89%, while in untreated grains resulted in 100% mortality. Silva et al. (2016) evaluated the were 2.08%, 1.63%, and 2.03%, for 2nd and 5th instars and toxicity of ozone (750.8 ppmv) against R. dominica adults adults, respectively (Fig. 4A). The moisture percentage in on wheat kernels and found that ozone exposure from 8.69 to ozone-treated grains were 8.67%, 8.14%, and 9.01%, while 13.08 h and from 11.28 to 18.11 h caused mortality rates of in untreated grains were 9.64%, 9.64%, and 9.36% for 2nd 50% and 95%, respectively. Subramanyam et al. (2017) also and 5th instars and adults, respectively (Fig. 4B). The ash recommended that higher ozone doses (200 and 400 ppmv) percentage in ozone-treated grains were 0.41%, 0.79%, are required to control R. dominica adults and reported that and 0.81%, while in untreated grains were 0.35%, 0.99%, adult mortality after one day were 67% and 42%, respec- and 0.65% for 2nd and 5th instars and adults, respectively tively. Gad et al. (2021b) demonstrated that treatment with (Fig. 4C). The carbohydrate percentage in ozone-treated 600 ppmv (1.2 g/m ) of ozone caused complete mortal- grains were 72.47%, 72.26%, and 71.72%, while in untreated ity of C. maculatus adults at all exposure times (0.5–5 h) grains were 72.56%, 71.60%, and 71.90% for 2nd and 5th after 5 d. Similar results obtained as larval mortality of E. instars and adults, respectively (Fig. 4D). The fiber percent - cautella, P. interpunctella, T. granarium and T. castaneum age in ozone-treated grains were 2.49%, 2.83%, and 2.50%, increased as concentration and exposure periods increased. while in untreated grains were 2.09%, 2.99%, and 2.25%, Complete mortality was observed after 8 h. The LT50–99 for 2nd and 5th instars and adults, respectively (Fig. 4E). values of ozone gas against the larvae decreased as concen- The protein percentage in ozone-treated grains were 14.11%, tration increased. Caterpillars were more sensitive to O than 14.54%, and 14.07%, while in untreated grains were 13.24%, grubs. Data also showed that the effective effect of ozonation 13.15%, and 13.81%, for 2nd and 5th instars and adults, towards the four larval species indicated that not all insects respectively (Fig. 4F). had the same sensitivity to ozone gas (Ghazawy et al. 2021). 1 3 Applied Entomology and Zoology Fig. 4 Effect of ozone gas on the quality of wheat kernels. A fat; B moisture; C ash; D carbohydrate; E fiber; and F protein Adult emergence of T. granarium was decreased in adult emergence percentages of T. granarium adults. Xinyi treated wheat kernels with all tested ozone concentrations et al. (2019) found that treatment with ozone at 0.42 g/ after 60 days of treatment. The results revealed 100% m suppressed adult progeny production of R. dominica reduction in Fl adult emergence of T. granarium at the in wheat. Gad et al. (2021a) mentioned that the number highest concentration of ozone (1200 ppmv). Similar find- of progeny of C. maculatus and C. chinensis was signifi- ings have been recorded by Bonjour et al. (2011) where a cantly decreased in all ozone concentrations after 45 d of 70 ppmv ozone treatment for 4 d caused a progeny reduc- treatment. tion of 100% of Oryzaphilus surinamensis (L.) and 98% This study also demonstrated that the wheat kernels of T. castaneum. Moreover, Abd El-Ghaffar et al. (2017) were protected for 60 days from weight loss brought found that increasing the gas concentration decreased the on by T. granarium by ozone treatment at the highest 1 3 Applied Entomology and Zoology Funding Open access funding provided by The Science, Technology concentration of 1200 ppmv. Our results are in line with & Innovation Funding Authority (STDF) in cooperation with The Pleijel and Uddling (2012) who observed that ozone less- Egyptian Knowledge Bank (EKB). This research did not receive any ens the typical weight loss of wheat kernels. Gad et al. specific grant from funding agencies in the public, commercial, or not- (2021a) reported similar findings, stating that all ozone for-profit sectors. treatments decreased the weight loss of treated cowpea Declarations seeds in comparison to untreated seeds. According to Dong et al. (2022), 1440 min of ozone exposure at 700 Conflict of interest The authors declare no conflict of interest. ppmv completely controlled all life stages of R. dominica and T. castaneum insects in barley without having a nega- Open Access This article is licensed under a Creative Commons Attri- bution 4.0 International License, which permits use, sharing, adapta- tive effect on the weight of the grain. tion, distribution and reproduction in any medium or format, as long Chemical analysis of infested wheat kernels treated at as you give appropriate credit to the original author(s) and the source, a concentration of 1200 ppmv ozone observed no signifi- provide a link to the Creative Commons licence, and indicate if changes cant changes in fat, moisture, ash, carbohydrate, fiber, and were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated protein content compared with infested wheat kernels that otherwise in a credit line to the material. If material is not included in were not treated with ozone. Changes in the nutrient con- the article's Creative Commons licence and your intended use is not tents of treated wheat kernels with ozone were supported permitted by statutory regulation or exceeds the permitted use, you will by the results of Wang et al. (2008), who found that the need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://cr eativ ecommons. or g/licen ses/ b y/4.0/ . protein content of ozone-treated corn is lower than that of untreated corn; this finding indicated that protein was destroyed by ozonation, thereby influencing the nutritional References value of the corn. Gozé et al. 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Applied Entomology and Zoology – Springer Journals
Published: May 1, 2023
Keywords: Khapra beetle; Ozone; Mortality; Adult emergence; Weight loss
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