Laboratory Evaluation of Indigenous and Commercial Entomopathogenic Nematodes against Red Palm Weevil, Rhynchophorus ferrugineus (Coleoptera: Curculionidae).

The red palm weevil (RPW) is a significant threat to date palms. Conventional pest control has been ineffective. This study aims to evaluate entomopathogenic nematodes (EPNs) indigenous to Saudi Arabia and commercial against RPW. We used 33 soil samples collected from four areas of Saudi Arabia. The indigenous EPNs were isolated and cultured using an insect baiting method to obtain infective juveniles. Pathogenicity bioassays were conducted against different stages of RPW, including eggs, larvae, and adults. The bioassay was performed using all the isolates at 1 × 106 IJ/mL. Distilled water was used as a control. The results revealed that only 9.09% of soil samples contained positive EPNs. Through DNA sequencing analysis, the positive samples were identified as indigenous isolates belonging to Heterorhabditis indica and Steinernema carpocapsae EPN species. In pathogenicity tests, 90% mortality of RPW eggs was observed after five days. Similar mortality trends were seen in RPW larvae and adults, with 90% mortality recorded after ten days for all the EPN treatments. Mortality increased with the duration of post-EPN inoculation exposure. The 1 × 106 IJ/mL concentrations of EPN effectively killed various stages of RPW in the laboratory. More research is needed to test EPNs against RPW in the field.

A winning formula: sustainable control of three stored-product insects through paired combinations of entomopathogenic fungus, diatomaceous earth, and lambda-cyhalothrin.

This research aimed to assess the effectiveness of Metarhizium robertsii, diatomaceous earth (Protect-It), and lambda-cyhalothrin, for the long-term protection of stored wheat against three destructive grain insect pests, Rhyzopertha dominica, Tribolium castaneum, and Trogoderma granarium. Different treatments were applied, both alone and in paired combinations in laboratory and persistence trials. Single treatments exhibited significantly lower mortality rates in comparison to the paired treatments for all tested insect species. Among the single treatments, lambda-cyhalothrin (Lamb) resulted in significantly higher mortality rates in laboratory trials, followed by diatomaceous earth (DE) and M. robertsii (Mr), with insignificant differences between Mr and DE. Evidently, DE exhibited the highest persistence after 120 days of storage for all insect species and initial exposures, although variations in mortality rates among treatments were mostly insignificant. Overall, the most effective treatment in terms of mortality in laboratory, and persistence trials, and progeny production was DE + Lamb, followed by Mr + Lamb, and Mr + DE for all tested insect species. In general, the most susceptible insect species was R. dominica, followed by T. castaneum and T. granarium. This research highlights the effectiveness of M. robertsii, DE, and lambda-cyhalothrin in providing prolonged protection of stored wheat against all the examined grain insect species.

Evaluating the Compatibility of Spinosad and Alpha-Cypermethrin for Controlling Six Insect Pests Infesting Stored Wheat.

The deterioration of stored wheat due to pest infestations is a significant concern, with pests like Rhyzopertha dominica, Tribolium castaneum, Cryptolestes ferrugineus, Sitophilus oryzae, Oryzaephilus surinamensis, and Trogoderma granarium being major contributors. This study examined the efficacy of spinosad and alpha-cypermethrin, individually and in combination, against these pests under laboratory conditions. Spinosad was tested at two concentrations (0.05 and 0.1 mg/kg), while alpha-cypermethrin was applied at 0.05 mg/kg. The combined application of both insecticides led to significantly higher pest mortality compared to single treatments. Importantly, all treatments caused substantial pest mortality and exhibited the ability to suppress pest progeny production over time, as observed in both laboratory and persistence trials. Among the various treatment combinations, the joint application of 0.1 mg/kg spinosad and 0.05 mg/kg alpha-cypermethrin emerged as the most effective, resulting in elevated mortality and a marked reduction in pest progeny. Rhyzopertha dominica exhibited the highest susceptibility among the pests, followed by S. oryzae, T. castaneum, C. ferrugineus, O. surinamensis, and T. granarium. The remarkable performance of the joint action of alpha-cypermethrin and spinosad at low doses highlights this combination as an efficacious approach for safeguarding stored grain against these destructive insect pests, warranting further exploration.

Natural Warriors against Stored-Grain Pests: The Joint Action of Beauveria bassiana and Steinernema carpocapsae.

Tribolium castaneum, Trogoderma granarium, Oryzaephilus surinamensis, Sitophilus oryzae, Rhyzopertha dominica, and Cryptolestes ferrugineus are all major pests of stored grains. In this study, the efficiency of single and joint applications of the entomopathogenic nematode (EPN) Steinernema carpocapsae at two different doses (50 and 100 IJs cm-2) and the entomopathogenic fungus (EPF) Beauveria bassiana for the management of the aforementioned pests was estimated. At single treatments, both doses of S. carpocapsae caused higher mortality rates to all six pest species compared to B. bassiana. The combined treatment of EPF and EPN resulted in higher mortality compared to single treatments. Mortality was strongly influenced by the exposure interval and the application dose of the EPN at both single and combined treatments. Maximum mortality was observed for the application of the combined treatment at the high dose of S. carpocapsae and B. bassiana. Among the different insect species tested, the maximum mortality rate was observed for R. dominica (96.62%), followed by S. oryzae (90.48%), T. castaneum (87.23%), C. ferrugineus (76.05%), O. surinamensis (70.74%), and T. granarium (57.71%). The outcomes of this study demonstrate the potential of utilizing specific combinations of EPF and EPN as effective natural enemies against stored-grain pests.

Influence of natural and non-natural diets on the fitness and rearing of Pectinophora gossypiella Saunders.

In order to develop integrated management approaches for Pectinophora gossypiella, basic studies are crucial. The two-sex life table is the most important tool for describing the fitness and population parameters of both sexes (male and female) of an insect, while the traditional life table only explains the female sex of an insect. However, no study has reported on the biology of P. gossypiella using two-sex life table tools. Therefore, this study explains the rearing dynamics of P. gossypiella on a cotton seed-based artificial diet and a natural diet (mature cotton bolls). According to the results, the oviposition period of P. gossypiella was recorded to be longer on the artificial diet (9.07 ± 0.24) compared to the natural diet (7.40 ± 0.11). The total fecundity of P. gossypiella was greater on the artificial diet (125.94 ± 3.06) in comparison to the natural diet (60.37 ± 1.10). The population parameters, including intrinsic rate of increase, finite rate of increase, gross reproductive rate, and net reproductive rate of P. gossypiella were highest on the artificial diet in comparison to the natural diet. This study concluded that the cotton seed-based artificial diet was most suitable for the rearing of P. gossypiella. In the future, P. gossypiella may be studied in depth in light of the findings in this study.

Treatment of Four Stored-Grain Pests with Thiamethoxam plus Chlorantraniliprole: Enhanced Impact on Different Types of Grain Commodities and Surfaces.

An insecticide containing the neonicotinoid thiamethoxam + the diamide chlorantraniliprole was evaluated against adults of Rhyzopertha dominica, Tribolium castaneum, Trogoderma granarium, and Sitophilus oryzae under laboratory bioassays both on freshly treated grain as well as on treated grain stored over 90 days for its persistence in efficacy. In laboratory bioassays, the insecticide was applied on wheat, maize, or rice at four doses, while in persistence bioassays on wheat at the same doses. Mortality and progeny were assessed in both laboratory and persistence bioassays. After 14 days of exposure, S. oryzae exhibited 100% mortality on all three commodities at the highest dose, while R. dominica showed complete mortality on wheat or rice and T. castaneum on wheat. For a period of 90 days, S. oryzae exhibited 42.69% mortality, followed by R. dominica (35.26%), T. castaneum (27.08%), and T. granarium (18.63%) at the highest dose. Progeny was successfully suppressed in all cases of complete mortality in laboratory bioassays and for S. oryzae for 90 days in persistence bioassays. Laboratory trials were also performed on plywood, concrete, ceramic tile, and steel at one dose. The highest mortality was observed on steel, followed by concrete, ceramic tile, and plywood for all insect species tested. This study demonstrates that thiamethoxam + chlorantraniliprole is effective against the tested species depending on exposure, storage period, surface, commodity, and dose.

The Potential of Two Entomopathogenic Fungi and Enhanced Diatomaceous Earth Mixed with Abamectin: A Comprehensive Study on Mortality, Progeny Production, Application Method, and Surface Application against Tribolium castaneum.

This study determined the efficacy of Beauveria bassiana (Bals. -Criv.) Vuill., Metarhizium anisopliae (Metchnikoff) Sorokin, and diatomaceous earth mixed with abamectin (DEA) alone and in their combinations for the integrated management of larvae and adults of Tribolium castaneum (Herbst) from three field populations of Pakistan (Multan, Rawalpindi, and Rahim Yar Khan) and one laboratory population (Faisalabad). Treatments were applied on three surfaces, namely, viz. steel, concrete, and jute bags, implementing two application methods, dusting and spraying. The combined treatments were more effective in comparison with single treatments for both larvae and adults. Overall, the highest mortality rates were recorded in the Faisalabad population, followed by the Rehaim Yar Khan, Rawalpindi, and Multan populations. Progeny production was suspended 21 days after exposure to the combined treatment of DEA and both fungi in all populations except Rawalpindi. Larvae were found to be more susceptible than adults in all treatments and intervals. Dusting was more efficient than spraying for both larvae and adults and for all the populations studied. The present study provides a wholistic understanding of the impact of different factors on the success of the combined treatments using DEA and entomopathogenic fungi, supporting their use as surface treatments.

Acoustic Comparisons of Red Palm Weevil (Rhynchophorus ferrugineus) Mortality in Naturally Infested Date Palms after Injection with Entomopathogenic Fungi or Nematodes, Aluminum Phosphide Fumigation, or Insecticidal Spray Treatments.

Red palm weevil (RPW) management is important to the economic success of date palm agriculture. Monitoring with acoustic sensors was conducted in naturally infested trees in date palm orchards for six months after treatments with entomopathogenic fungi (Beauveria bassiana and Metarhizium anisopliae), entomopathogenic nematodes (Steinernema carpocapsae), aluminum phosphide, emamectin benzoate, or fipronil to evaluate their efficacy in an integrated pest management treatment vs. a distilled water injection. Reductions in the mean rates of RPW sound impulse bursts over time after treatment were used as indicators of RPW mortality. Entomopathogenic fungi and nematodes, aluminum phosphide, and emamectin benzoate were the most effective treatments, reducing RPW impulse burst rates within 2-3-months to levels indicating absence of infestation. However, when applied as a spray, fipronil had only a minor effect. The results indicate that treatments utilizing entomopathogenic fungi or nematodes can beneficially manage RPW in palm orchards and can help to limit treatments that may induce insecticide resistance or cause human and environmental harm. Furthermore, the use of an acoustic sensor can be beneficial in monitoring the activities of insect borers inside the tree trunk.

Development of Insecticide Resistance in Field Populations of Onion Thrips, Thrips tabaci (Thysanoptera: Thripidae).

The present study evaluated insecticide resistance in field populations of onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), collected from eight different onion-growing regions of Punjab, Pakistan. These field-collected populations were assessed for resistance development against eight commonly used active ingredients including deltamethrin, lambda-cyhalothrin, imidacloprid, acetamiprid, spinosad, spinetoram, cypermethrin, and abamectin. In leaf dip bioassays, T. tabaci adults showed varied levels of resistance towards different insecticides. Moderate or high levels of resistance to deltamethrin (58-86 fold), lambda-cyhalothrin (20-63 fold), and cypermethrin (22-54 fold) were observed in T. tabaci field populations. There were very low to moderate resistance levels to imidacloprid (10-38 fold), acetamiprid (5-29 fold), and abamectin (10-30 fold). The lowest levels of resistance were detected in thrips exposed to spinosad (3-13 fold) and spinetoram (3-8 fold). Insecticide resistance levels varied among populations collected from various geographic locations, but all populations exhibited elevated levels of resistance to deltamethrin. Thrips tabaci populations with higher resistance levels were most commonly found from the southern part of Punjab, Pakistan. Our findings revealed that spinosyns could be used as alternatives to conventional insecticides for the successful management of T. tabaci in onion fields.

Field Evaluation of Promising Indigenous Entomopathogenic Fungal Isolates against Red Palm Weevil, Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae).

The rate of the sounds (i.e., substrate vibrations) produced by the movement and feeding activity of red palm weevil (RPW) pest infestations in a date palm tree was monitored over time after trees were separately treated with injection of entomopathogenic fungal isolates, Beauveria bassiana and Metarhizium anisopliae, or water treatment as the control. The activity sensing device included an accelerometer, an amplifier, a digital recorder, and a signal transmitter that fed the data to a computer that excluded background noise and compared the rates of bursts of movement and feeding sound impulses among treated trees and controls. Observations were made daily for two months. The rates of bursts were representative of the feeding activity of RPW. The unique spectral pattern of sound pulses was typical of the RPW larval feeding activity in the date palm. The microphone confirmed that the same unique tone was produced in each burst. Two months after fungal injection, the RPW sound signal declined, while the RPW sound signal increased in the control date palms (water injection). The mean rates of bursts produced by RPW decreased to zero after the trees were injected with B. bassiana or M. anisopliae compared with the increased rates over time in the control treatment plants.

Silencing of vitellogenin gene contributes to the promise of controlling red palm weevil, Rhynchophorus ferrugineus (Olivier).

Red palm weevil [Rhynchophorus ferrugineus (Olivier)], is native to South Asia and expanding its distribution range globally. Recent invasions of red palm weevil around the world, including Saudi Arabia, has become a global constraint for the production of palm species. Although, several control measures have been tested, none of them seemed successful against this invasive species. Therefore, we focused on silencing the reproduction control gene vitellogenin (Vg) based on RNA interference (RNAi) strategy for its possible application in the management of R. ferrugineus. The Vg is a major yolk protein precursor critical for oogenesis. To do this, fat body transcriptome of R. ferrugineus female adults was sequenced, which provided partial Vg gene transcript (FPKM 5731.60). A complete RfVg gene transcript of 5504 bp encoding 1787 amino acids was then sequenced using RCAE-PCR strategy and characterized. Phylogenetic analysis suggested that RfVg has closer ancestry to the coleopteran insects. The RfVg-based RNAi significantly suppressed the expressions of Vg gene. The 15, 20 and 25 days post-injection periods suppressed Vg expressions by 95, 96.6 and 99%, respectively. The suppressed Vg expressions resulted in the dramatic failure of Vg protein expression, which caused atrophied ovaries or no oogenesis and ultimately eggs were not hatched. These results suggest that knockdown of Vg gene involved in R. ferrugineus reproduction is a promising target for RNAi-based management of R. ferrugineus.

Prediction of survival ratios of Cadra cautella (Lepidoptera: Pyralidae) different life stages after treated with ultraviolet radiation in dates.

Date palm, is a tree of economic importance which is grown around the world, including Saudi Arabia. Its fruit is nutritious and possesses medicinal benefits. Almond moth, is a serious date fruits pest in the field as well as in the storage and causes severe economic losses. In the given research, ultraviolet radiation type B (UV-B, 315 nm) harmful effects were evaluated against all developmental stages of C. cautella. One and 3-d-old eggs, 12 and 18-d-old larvae, 1-d and 6-d-old pupae, and 1-d-old adults, were exposed to UV-B for different intervals. Eggs were exposed for 0-30 min and 0% hatchability was achieved both for 1-d and 3-d-old eggs after 30 min. The larvae were exposed for 6-24 h, and after 24 h, mortality was 100 and 97% for 12 and 18-d-old larvae, respectively. Similarly, the pupae were exposed for 0-30 h, and 100% mortality was achieved after 30 h for 1-d-old pupae. Furthermore, none of the 6-d-old pupae emerged as an adult after 12 h of exposure. When adults were exposed for 1-4 d, no mortality was observed; however, UV-B reduced fecundity and hatchability in the treated adults. The susceptibility order was as follows: eggs > larvae > pupae > adults. Several uncharacteristic behaviors of C. cautella were noted, such as females depositing eggs openly on food items and containers, mature larvae exiting from food, larvae starting to wander for pupation, and pupation occurring typically outside the food. The application of UV-B could be an effective management strategy because all developmental stages of C. cautella were susceptible to UV-B that might be helpful to protect the dates from C. cautella infestation.

Comparative efficacy of CO2 and ozone gases against Ephestia cautella (Lepidoptera: Pyralidae) larvae under different temperature regimes.

Comparative efficacy of three different modified atmospheres: 100% CO2, 75% CO2 + 25% N2, and 22 ppm ozone were examined against larval mortality of the almond moth, Ephestia cautella (Walker) (Lepidoptera: Pyralidae) at temperature regimes of 25°C and 35 ± 2°C and 60 ± 5% relative humidity, and 9:15 dark and light. Wandering young larval instars, which are fast growing, large enough in size and considered as more tolerant to modified atmosphere, were collected directly from the rearing culture, placed inside pitted date fruits of vars.: "Khudri," "Ruziz," and "Saqie," were treated with aforementioned gases for 24, 48, and 72 h. The immediate and delayed larval mortality was recorded after each exposure timing. Ozone possessed the strongest fumigant toxicity causing 100% mortality with all varieties, at 25 and 35°C after 24 h exposure and was more effective than 75% CO2 that caused 83 and 100% immediate mortality with variety ruziz at 25 and 35°C, respectively. Extending the treatments exposure time to 72 h, 100% mortality was recorded by exposing larvae to any of the studied gases at 25 and 35°C. These results suggest that gases and temperature used in this study can be effectively used to control E. cautella in dates and stored grains.