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Induction of anhydrobiosis and storage stability of entomopathogenic nematodes are influenced by moisture availability. Decreasing moisture content in diatomaceous earth (DE) pellets containing theSteinernema glaseriNJ-43 strain and its effect on survival time and infectivity of the nematode were determined. Pelletisation was performed in a vortex mixer, using DE Celite® 209 as the desiccant material. Pellets were stored at room temperature (23 ± 2°C) and high relative humidity (96–100%). Nematode survival and infectivity against last instar greater wax moth,Galleria mellonella,were tested daily. Initial average and average equilibrium moisture content in pellets were 66.7% and 13.6%, respectively, and the infective juveniles mean survival time was 8.8 days. A moisture transfer model based on diffusion and evaporation was evaluated to predict moisture fluctuations within the pellets. We concluded that 84% of variation inS. glaseriinfectivity onG. mellonellalarvae was explained by the survival of the nematode, whereas 52% of variation inS. glaserisurvival was explained by the loss of moisture from the pellets. The moisture transfer model achieved 78% reliability in predicting moisture content and fluctuations. Therefore, the mechanisms of moisture diffusion and evaporation from the surface to the surrounding atmosphere contribute significantly to moisture loss from the pellets. [ABSTRACT FROM AUTHOR]

Although their control is based on chemical products, the infestations by ticks (Ixodes scapularis Say) are causing great losses and damages in the livestock production worldwide. In this study, the survival of the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema carpocapsae, and Steinernema websteri in vegetal oil suspension at concentrations of 13% and 33% and their effectiveness to control ticks at concentrations of 50 ± 5 and 100 ± 10 nematodes in oil suspensions of Cymbopogon citratus, Pelargonium sp, Juniperus virginiana, Rosa sp, and Mentha piperita were evaluated in lab conditions. In field conditions, the Lethal Concentration (LC90) of S. websteri in oil suspensions of J. virginiana and C. citratus in dogs infested with ticks was evaluated. In the laboratory, it was found that an oil emulsion of C. citratus and J. virginiana at 13% maintained the survival of S. carpocapsae, H. bacteriophora, and S. websteri from 55% to 60% for a period of 96 hr. The combination of the S. websteri nematode with 50 or 100 nematodes in oil emulsions of J. virginiana at 33% presented a control effectiveness of 80–100% in adult ticks 24 hr post-application. In field, the LC90 of 119 juveniles of S. websteri in oil emulsions of J. virginiana at 33% on domestic dogs presented an accumulated and a control effectiveness of 89% after 96 hr post-application. The combined application of J. virginiana and S. websteri could be a good alternative for the control of ticks. It was observed that the time of contact and the type of vegetable oil were crucial factors to increase the effectiveness of control.

Applications of infective juveniles (IJ) of entomopathogenic nematodes (EPN) formulated in pellets are still limited. This is principally due to limited advances in the technology of formulation. We aimed to develop a new method of mechanical formulation through material flow and to analyse its effect on the survival time of encapsulated EPN by varying the granular materials, the components of the aqueous suspension, the age of the nematodes and by applying a surface coating (C) to the pellet. Three-day-old and two-month-oldSteinernema glaseriIJ were encapsulated with different proportions of diatomaceous earth (DE) and attapulgite clay (AC). The aqueous suspension containing the nematodes was prepared with double distilled water (DDW), varying proportions ofOpuntia ficus-indicamucilage (OM) or gelatin (GL), and a sunflower oil surface treatment. The pellets were stored at an average room temperature of 23 ± 6°C. The best results were obtained with the following proportions: 100DE:0AC and 50DE:50AC, using the OM suspension, three-day-old nematodes and a surface C, which resulted in an average of 14 days survival time. These results confirmed that the nematodes do not die during mechanical encapsulation and that the age of the IJ as well as the loss of moisture during storage at room temperature were the factors that decreased the survival of encapsulated EPN. It was concluded that it is necessary to use neonate IJ and to reduce the moisture transfer rate in the granular structure in order to delay the desiccation of the encapsulated nematodes. [ABSTRACT FROM AUTHOR]

Phyllophaga spp. are a complex of edaphic insect pests that are present in the corn crops ( Zea mays ) in México, which are usually controlled with increasing dosages of broad-spectrum chemical insecticides. Several entomopathogenic nematode species can produce acceptable control levels of these larvae. However, the synergistic interaction between fungi and entomopathogenic nematodes (EPN) could improve the control of this insect. This study investigates the mortality of larvae of Phyllophaga vetula by the effect of the separate or combined application of the fungus Metarhizium anisopliae M1cog strain (Ma) and the nematodes Steinernema carpocapsae All strain (Sc) or Steinernema glaseri NJ-43 strain (Sg). In laboratory, dosages of 1 × 10 6 or 1 × 10 8  spores/larva and 250 infective juveniles were applied on medium or large size P. vetula larvae contained in vials with sterilized agricultural soil as the assay arena. The separate application of Ma did not kill any larvae, but Sg and Sc killed 40 and 80% of the larvae, respectively. However, the Ma and Sc combination had an important antagonistic interaction that decreased the mortality to 40%, but the combination Ma and Sg had a slight additive interaction that increased the mortality to 47%. The most determining factor in larvae mortality was the nematode used, with Sg as the species with best performance in 6 of the 12 treatments evaluated and with a maximum effectivity of 80% on medium-size larvae if combined with a low dosage of Ma. The combined application of an entomopathogenic fungus and EPN showed no consistent effects on the mortality percentage of P. vetula , mostly because the fungus was not isolated from Phyllophaga larvae., Phyllophaga spp. are a complex of edaphic insect pests that are present in the corn crops ( Zea mays ) in México, which are usually controlled with increasing dosages of broad-spectrum chemical insecticides. Several entomopathogenic nematode species can produce acceptable control levels of these larvae. However, the synergistic interaction between fungi and entomopathogenic nematodes (EPN) could improve the control of this insect. This study investigates the mortality of larvae of Phyllophaga vetula by the effect of the separate or combined application of the fungus Metarhizium anisopliae M1cog strain (Ma) and the nematodes Steinernema carpocapsae All strain (Sc) or Steinernema glaseri NJ-43 strain (Sg). In laboratory, dosages of 1 × 10 6 or 1 × 10 8  spores/larva and 250 infective juveniles were applied on medium or large size P. vetula larvae contained in vials with sterilized agricultural soil as the assay arena. The separate application of Ma did not kill any larvae, but Sg and Sc killed 40 and 80% of the larvae, respectively. However, the Ma and Sc combination had an important antagonistic interaction that decreased the mortality to 40%, but the combination Ma and Sg had a slight additive interaction that increased the mortality to 47%. The most determining factor in larvae mortality was the nematode used, with Sg as the species with best performance in 6 of the 12 treatments evaluated and with a maximum effectivity of 80% on medium-size larvae if combined with a low dosage of Ma. The combined application of an entomopathogenic fungus and EPN showed no consistent effects on the mortality percentage of P. vetula , mostly because the fungus was not isolated from Phyllophaga larvae.

The inhibitory effect of Chrysomya rufifacies (Macquart) and Cochliomyia macellaria (Fabricius) larval excretions-secretions (ES) on Staphylococcus aureus was determined using a portable colorimetric method without the need for any dedicated spectral instrument. Polystyrene 96 well microplates were used and 100 μl of the bacterial inoculum (5 × 105 CFU/ml) plus 100 μl of the dipteran exosecretions at different concentrations were added to each well. Subsequently, 50 μl of a 1% solution of the triphenyl tetrazolium chloride stain was added to each well to determine the bacterial viability. The color development in each well was measured with the ImageJ software S. aureus was exposed to different concentrations of the ES of both species individually. At a concentration of 800 ppm ES of C. rufifacies or Co. macellaria, bacterial growth was inhibited 97.45 ± 1.70% and 82.21 ± 1.88%, respectively. As expected, exposure to a lower concentration (i.e., 50 ppm) was less inhibitory (C. rufifacies ES, 77.65 ± 4.25% and Co. macellaria ES, 43.54 ± 4.63%). This study demonstrates for the first time the bactericidal activity of C. rufifacies and Co. macellaria ES against S. aureus. This finding is promising as it could result in the identification and synthesis of proteins capable of suppressing pathogen development in wounds. Additionally, the proposed method can simplify the use of expensive laboratory instruments for antimicrobial activity determination., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

An alternative control method to the use of chemical insecticides against soil dwelling insect pests is the application of entomopathogenic nematodes formulated in alginate beads for enhanced shelf life. The aim was to compare the benefit on nematode survival and infectivity of: (i) pre-conditioning of juveniles, and (ii) coating of alginate beads. The nematodes Steinernema glaseri , Steinernema carpocapsae, and Heterorhabditis bacteriophora were reproduced in last instar larvae of the wax moth Galleria mellonella and the infective juveniles emerged were subjected to two capture treatments: white traps and plaster of Paris, the latter was utilized as a pre-conditioning treatment. A total of 1,000 infective juveniles were formulated in each sodium alginate bead with or without an alginate coating. The beads were stored at 23 ± 3°C and a bidistilled water suspension of nematodes was set as a control. The survivorship of these formulates and their infectivity on Galleria mellonella larvae were evaluated every 7 d post-formulation (dpf). In beads without pre-conditioning juveniles nor coating, Steinernema carpocapsae had the higher survival rate (58.8%), the longest survival time (28 dpf) and the higher infectivity (100%) as well. Pre-conditioning improved the survival and infectivity of Steinernema glaseri by 22.5% at 21 dpf and 70.0% at 14 dpf, respectively. Alginate coating increased survival (21.7%) and infectivity (95%) of Heterorhabditis bacteriophora for up to 28 dpf. The combination of pre-conditioning and coating slightly favored the survival (10% for up to 14 dpf) of Steinernema glaseri and infectivity (100% for up to 35 dpf) of Steinernema carpocapsae . Non pre-conditioned Steinernema carpocapsae formulated in uncoated beads was the combination with better performance in survivorship (58.8%) and infectivity (100%) at 28 dpf. It was concluded that non pre-conditioned Steinernema carpocapsae formulated in uncoated beads was the combination with better survivorship and infectivity., (© The Society of Nematologists.)

Objective: To determine larvicidal activity of the essential oil, hydrolat and botanical extracts derived from leaves of Pseudocalymma alliaceum on mosquito larvae of Culex quinquefasciatus., Methods: Groups of twenty larvae were used in the larvicidal assays. The mortality, relative growth rate, the larval and pupal duration and viability was estimated. The essential oil was analyzed by solid phase microextraction using gas chromatography coupled to mass spectrometry., Results: Essential oil at 800 ppm showed larvicidal activity at 24 h with lethal values of LC50 and LC90 of 267.33 and 493.63 ppm. The hydrolat at 20% and 10% on 2nd stage larvae showed 100% effectiveness after 24 h. The aqueous extract at 10% had a relative growth index of 0.58, while the ethanolic and methanolic extract obtained values of 0.76 and 0.70 and control reached 0.99. Larvae treated with 10% of methanol, ethanol and aqueous extract showed a reduction in larval duration of 5.00, 2.20 and 4.35 days; ethanol extract at 1% provoke decrease of 2.40 days in the development and exhibited an increment of 3.30 days when treated with 0.01%. Aqueous, ethanol and methanol extracts at 10% reduced in 6.15, 3.42 and 5.57 days pupal development. The main compounds were diallyl disulfide (50.05%), diallyl sulfide (11.77%) and trisulfide di-2-propenyl (10.37%)., Conclusions: The study demonstrated for the first time, the larvicidal activity of the essential oil and hydrolat of Pseudocalymma alliaceum; aqueous, ethanol and methanol extracts inhibited the normal growth and development of mosquito larvae, prolonging and delaying larval and pupal duration., (Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)