Evaluation of Metarhizium rileyi Farlow (Samson) impregnated with azadirachtin and indoxacarb against Helicoverpa armigera (Hubner)

Background Entomopathogenic fungi are the most versatile having a wide host range, capable of infecting insects at different developmental stages. In the present study, Metarhizium rileyi, at the concentrations of 102, 103, 104, 105, 106, 107 and 108 conidia/ml and sub-lethal concentrations of azadirachtin (1.02 and 1.53 ppm) and indoxacarb (0.72 ppm) were evaluated against the 1st, 2nd, 3rd, 4th and 5th larval instars of Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) under laboratory conditions. Results M. rileyi applied at 106 conidia/ml caused a maximum mortality of 83.33 and 80.00% of 1st and 2nd larval instars of H. armigera, respectively. The maximum mortality of 3rd, 4th and 5th larval instars of H. armigera with 108 conidia/ml of M. rileyi was 83.33, 76.67 and 53.33%, respectively. When M. rileyi blended with azadirachtin at 1.02 ppm, the highest mortality rate of 86.21% at 106 conidia/ml against 2nd instar larvae was resulted. Similarly, M. rileyi applied at 108 conidia /ml mixed with azadirachtin (1.53 ppm) showed 89.66% mortality of 3rd instar larvae. The 2nd instar larvae treated with M. rileyi at 106 conidia/ml, mixed with indoxacarb (0.72 ppm), the corrected mortality rate was 82.14%. Concentration mortality response of 3rd instar larvae to M. rileyi blended with indoxacarb (0.72 ppm) was 85.71% at 108 conidia/ml. The median lethal concentration (LC50) values were 5.51 × 103, 1.86 × 104, 2.81 × 105 and 5.55 × 105 conidia/ml for 1st, 2nd, 3rd and 4th larval instars, respectively, after 7 days of treatment. M. rileyi when mixed with sub-lethal concentrations of azadirachtin (1.02 ppm) and indoxacarb (0.72 ppm) resulted LC50 values of 1.09 × 104 conidia/ml and 1.37 × 104 conidia/ml against 2nd instar larvae, respectively, after 24 hours. Similarly, M. rileyi mixed with sub-lethal concentrations of azadirachtin (1.53 ppm) and indoxacarb (0.72 ppm) resulted LC50 values of 3.12 × 108 and 3.06 × 105 conidia/ml against 3rd instar larvae, respectively, after 24 hours. The study revealed that the susceptibility of larvae decreased in case of large larval instars. Conclusions M. rileyi can be utilized as one of the component of Integrated Pest Management Program for the eco-friendly management of H. armigera. As the application of M. rileyi @ 107 conidia/ml alone or in combination with azadirachtin (1.02 and 1.53 ppm) or indoxacarb (0.72 ppm) resulted to the highest mortality.

Ocimum gratissimum essential oil improved the health, innate immunity and resistance to Aeromonas hydrophila infection in Pseudoplatystoma reticulatum

The essential oil of alfavaca (Ocimum gratissimum) contains important compounds, such as 1.8-cineol and eugenol. This study evaluated the effects of diets containing alfavaca essential oil on the zootechnical performance, plasma glucose, leukocyte respiratory activity, haematology, and intestinal histomorphometry in “cachara” (Pseudoplatystoma reticulatum) challenged with Aeromonas hydrophila. A total of 300 juvenile fish, with an average total length of 14.98 ± 0.28 cm and average weight of 18.84 ± 1.22 g, distributed in 20 tanks of 80 L, were fed twice a day with a diet containing essential oil at 0.5, 1.0, or 1.5% or without its inclusion in five replicates. After 45 days, blood collection and mid-intestinal bowel were sampled, before a challenge with Aeromonas hydrophila was performed. The fish supplemented with 1.0% of essential oil presented better weight gain, mean corpuscular volume, concentration of thrombotic and neutrophils, respiratory activity of leukocytes, and height of villi (p < 0.05) when compared to other groups. The cumulative mortality response was lower for fish fed 1.5% essential oil of alfavaca diets. Thus, the use of alfavaca essential oil is recommended in the diet of cachara catfish (P. reticulatum) at the level of up to 1.0% for 45 days for improvements in the zootechnical, haematological, and intestinal histomorphometric parameters.

Postharvest Fumigation of Fresh Citrus with Cylinderized Phosphine to Control Bean Thrips (Thysanoptera: Thripidae)

Bean thrips (BT), Caliothrips fasciatus (Pergande), is a pest of concern to certain countries that import fresh citrus fruit from California, USA. A series of laboratory-scale exploratory fumigations with phosphine at 4.9 ± 0.3 °C (mean ± 2 SD; x¯±2s) were conducted to evaluate the postharvest control of adult BT. Models of the duration–mortality response predicted ca. 99% mortality of BT populations when headspace concentrations of phosphine, [PH3], are maintained at levels ≥0.4 g m−3 (250 ppmv (µL L−1)) and ≤1.5 g m−3 (1000 ppmv (µL L−1)) for 12 h, with the duration representing the lower bound of the 95% confidence level (CL). Confirmatory fumigations, each lasting 12 h, were then conducted using BT-infested sweet oranges, Citrus sinensis (L.), at pulp temperature (T) ≤ 5 °C to corroborate the exploratory results. Three formulations of cylinderized phosphine were used: 1.6% phosphine by volume in nitrogen, VAPORPH3OS®, and ECOFUME®, all applied at two levels, ca. 1.5 g m−3 (1000 ppmv (µL L−1)), as well as 0.5 g m−3 (300 ppmv (µL L−1)). Collectively, across the formulations, an applied dose of ca. 1.5 g m−3 (1000 ppmv (µL L−1)) resulted in 0 survivors from 38,993 (probit 8.60, 95% CL; probit 9, 72% CL) treated BT, while an applied dose of 0.5 g m−3 (300 ppmv (µL L−1)) resulted in 0 survivors from 31,204 (probit 8.56, 95% CL; probit 9, 70% CL) treated BT. Results were discussed in the context of commercial and operational features of quarantine and pre-shipment (QPS) uses of phosphine to treat fresh fruit and, specifically, the control of BT in fresh citrus exported from California, USA, to Australia.

Concentration–Mortality Response of Mexican Populations Fall Armyworm (Lepidoptera: Noctuidae) to Commercial Formulations of Bacillus Thuringiensis

The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is the most economically important pest of maize (Zea mays L.) grown in Mexico. In order to identify biologically based management tactics for this pest, we determined the concentration–mortality response of 16 S. frugiperda populations collected in Mexico to two commercial formulations of Bacillus thuringiensis Berliner (Bt) (XenTari® [Bt subsp. aizawai] and DiPel® [Bt subsp. kurstaki]; Valent de México SA De CV, Jalisco, México). Laboratory bioassays established median lethal concentrations (LC50s) for each Bt formulation × population combination. We also determined mean larval weight 7 d after exposure and the number of larvae that reached the third instar of development. The populations were susceptible to both Bt formulations but they were, overall, more susceptible to Bt subsp. aizawai (XenTari) than to Bt subsp. kurstaki (DiPel). These results can serve as a reference to detect changes in S. frugiperda response to these Bt commercial products over time.

Comparison of Toxicological Bioassays for Whiteflies

Two Bemisia tabaci populations from Georgia and Florida, USA, were tested for their response to insecticides across different toxicological bioassay methods. Five insecticides in four Insecticide Resistance Action Committee (IRAC) groups (imidacloprid (4A), dinotefuran (4A), flupyradifurone (4D), pyriproxyfen (7C) and cyantraniliprole (28)), were evaluated against a water check. The routes of application to the plant used were either leaf drench or (systemic) root drench. The four different whitefly bioassay methodologies tested were two published IRAC methods, a clip cage method, and a new tube method. A split–split experimental design was used to assess any interactions between application route, bioassay method and insecticide treatment. Application route had no significant effect on efficacy. However, bioassay method affected overall whitefly mortality, with the dish method having reduced mortality compared to other methods, except for the clip cage method. High rates of cyantraniliprole, dinotefuran and flupyradifurone insecticides resulted in the highest incidence of adult whitefly mortality. Significant interactions relative to percent adult mortality were found between the insecticide and bioassay method for both populations assayed. The clip cage method was more sensitive in terms of dose mortality response followed by the cup and tube methods. The dish method was the least responsive to insecticide dose. Other interactions are discussed.

Synergism Between Phosphine (PH3) and Carbon Dioxide (CO2): Implications for Managing PH3 Resistance in Rusty Grain Beetle (Laemophloeidae: Coleoptera)

Strong resistance to phosphine (PH3) in the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Laemophloeidae: Coleoptera) poses a serious risk to stored-grain biosecurity. Resistant populations hold risk of surviving in PH3 fumigation, particularly in storage structure that limits achieving very high concentrations of PH3, demanding the need for alternative fumigation strategies. Cofumigation with PH3 and carbon dioxide (CO2) is one alternative approach that has the potential to be used widely. CO2 fumigation of adults of strongly PH3-resistant reference strain of C. ferrugineus, for 48 h, showed that the effective concentration (LC50) of CO2 was 30.99%. This 30% level of CO2 in combination with PH3 decreased the LC50 of PH3 from 6.7 mg/liter to 0.84 mg/liter, an eightfold increase in PH3 efficacy relative to PH3 fumigation in normal air. The LC99.9 decreased from 16.2 mg/liter to 5.8 mg/liter, a 2.8-fold increase in PH3 efficacy. Comparison of mortality response data of PH3 alone and the PH3 + CO2 mixture confirmed that CO2 enhances the toxicity of PH3 synergistically in addition to exerting its own toxicity. These results were validated against three independently field-derived strains of strongly resistant C. ferrugineus that confirmed that observed enhancement in toxicity with the PH3 + CO2 mixture was consistent, irrespective of differences in resistance phenotypes and inherent tolerance levels. Results of the current study provide further opportunities to develop new commercially viable strategy to control strongly PH3-resistant C. ferrugineus.

Potential Synergy between Spores of Metarhizium anisopliae and Plant Secondary Metabolite, 1-Chlorooctadecane for Effective Natural Acaricide Development

Date palm dust mites are important pests severely infesting valuable nutritious fruits (dates) of date palm. In search of an alternative to acaricides, joint action of Metarhizium anisopliae EBCL 02049 spores and 1-Chlorooctadecane was evaluated as a potential candidate for the management of Oligonychus afrasiaticus through natural products. In this regard, in vitro tests were performed to evaluate the interaction of M. anisopliae spores with multiple doses of 1-Chlorooctadecane (0.8, 1.6, 2.4, 3.2, and 4.0 mg/mL). Compatibility bioassay results evidenced from vegetative growth (77.7–84.40 mm), sporulation (5.50–7.30 × 106 spores/mL), and germination (96.70–98.20%), revealed that all the tested doses are compatible (biological index > 82) with the spores of M. anisopliae. The impact of combined treatment of spores with 1-Chlorooctadecane in different proportions (Scheme I, II, III, and IV) compared to their sole application against O. afrasiaticus was evaluated by concentration–mortality response bioassays. Results showed that all the combined treatments revealed high mortality compared to the sole application, which showed relatively slow mortality response over time. Toxicity recorded from Scheme IV combinations (80% 1-Chlorooctadecane: 20% Spores), exhibited strong synergistic interaction (joint toxicity = 713). Furthermore, potent interactions have overcome the host antioxidant defense at the final stage of infection by tremendously reducing catalase, and superoxide dismutase activities. These experiments demonstrated fungal–toxin joint synergistic interaction as a promising date palm dust mite management option.