The ability of the isolate VG008 of S. frugiperda granulovirus (SpfrGV) to enhance the infectivity of the isolate SfCOL of S. frugiperda multiple nucleopolyhedrovirus (SpfrMNPV) was evaluated on S. frugiperda larvae. Bioassays were performed with mixtures by using different proportions 90%:10% (M1), 95%:5% (M2) and 97.5%:2.5% (M3) of SfCOL:VG008, respectively. All mixtures showed higher insecticidal activity that SfCOL. The mixture M3 showed the highest enhancement of SfCOL reducing 11.40 times the Mean Lethal Concentration and 96 h in the Mean Time to Death. The enhancer activity of proteins derived from VG008 (GVPs) were also evaluated in mixture with SfCOL. The GVPs increased 27% larval mortality caused by SfCOL and damaged the peritrophic membrane of S. litura larvae, suggesting that the key point in this enhancing activity is the initial step of the larva colonization, the midgut infection. M3 was formulated and evaluated under greenhouse conditions in maize plants using different doses. The highest efficacy was obtained with the highest dose of M3 (8 × 1011 OBs/ha), which was similar to that found when formulated SfCOL was applied using an approximately twofold higher dose. The viral mixture M3 was selected as the active ingredient for developing a new biopesticide for a more efficient management of the pest in the field.

The tomato leafminer Tuta absoluta (Meyrick 1917) (Lepidoptera:Gelechiidae) is a devastating pest, causing losses of up to 100%. An interesting tool for its control is the use of the insect specific viruses of Baculoviridae family. Despite its high potential, its use on T. absoluta larvae has been poorly studied. In this work, Colombian granuloviruses VG013 and VG003 isolated from T. absoluta and Tecia solanivora (Lepidoptera:Gelechiidae) larvae sampled in tomato and potato crops, respectively, were morphologically, biologically and molecularly characterized. Occlusion bodies showed ovoid shape containing one nucleocapsid. Restriction endonuclease analysis revealed a pattern similar to Phthorimaea operculella granulovirus and bioinformatics studies showed that both isolates are variants of that baculovirus specie. Similar mean lethal concentrations (LC50) on T. absoluta larvae were estimated for both viruses, although VG013 exhibited shorter mean time to death than VG003. The latter reached a higher OBs yield in comparison with VG013. These results demonstrated an interesting potential of evaluated betabaculoviruses to control T. absoluta populations and pointed key features to its use under field conditions.

BACKGROUND: The hornworn Erinnyis ello is the major pest of natural rubber crops in Colombia, mainly controlled using toxic chemical insecticides. The use of E. ello Betabaculovirus is an environmentally sustainable alternative for its control. The aim of the present work was to characterize a prototype biopesticide formulation and evaluate its efficacy under different conditions. RESULTS: Quality control evaluations of formulated biopesticide revealed that all the parameters evaluated were under the permissible level. The lethal concentrations LC 50 and LC 90 of the biopesticide were 4.3 × 10 3 and 5.5 × 10 4 occlusion bodies (OBs) mL −1 , respectively. Biopesticide efficacies against second and fourth instar larvae under greenhouse conditions were higher than 80%. Evaluation of two application rates in a clonal garden resulted in 84% and 88% efficacy, comparable to that obtained with the chemical. The biopesticide in a commercial plantation showed efficacies between 74% and 82%. Biopesticide post-application persistence was estimated at least in 1 week under field natural conditions. Results allowed selection of the lowest evaluated dose (1 × 10 11 OBs ha −1 ) as the basis for further field evaluations. CONCLUSION: Formulated ErelGV showed high efficacy to control the hornworm in rubber crops and high potential to be included in integrated pest management programs, thus it could be an interesting alternative to replace agrochemicals.

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Wettable powder formulations by microencapsulation of viral occlusion bodies (OBs) of both Spodoptera frugiperda multiple nucleopolyhedrovirus from Colombia (SfCOL) and a genotypic variant (SfCOL-A) were evaluated for controlling the fall armyworm S. frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) in maize crops. Microencapsulation preserved OBs activity after three months of storage at 35ºC, where insecticidal activity loss was not greater than 12%. Additionally, the formulation protected the OBs against inactivation caused by UV-B radiation, retaining its insecticidal activity after 6 hours of UV laboratory exposure, in contrast to unformulated viral suspensions, which presented an Original Activity Remaining (OAR) between 12.1 and 50%. Under greenhouse conditions, the insect mortality was greater than 80% with microencapsulated viruses. In field trials, treatments reduced the percentage of damaged plants to levels below the economic injury level (35%) when the formulated and unformulated virus were applied at 8x1011 OBs/ha (800g/ha) dose, while the damage in the control treatment was close to 60%. Microencapsulation of SfCOL and SfCOL-A OBs provides useful advantages related to half-life and photostability.

Virus-based biopesticides are effective biocontrol agents of crop insect pests. Development of suitable formulations and production processes are necessary to obtain high-quality products easily adopted by farmers. A detailed unit operation study was carried out for the production process of a Phthorimaea operculella granulovirus-based biopesticide to control the tomato leafminer, Tuta absoluta, one of the most important pests affecting this crop. Physicochemical, microbiological, and insecticidal parameters were implemented in the process and applied to the finished product, and a scaling strategy was developed. A Quantitative Polymerase Chain Reaction (Q-PCR) technique was implemented to quantify viral concentrations in the active ingredient (5.34 ± 1.44 x10 9 Occlusion Bodies mL -1 ) and in the finished product (>1.6x10 9 OB mL -1 ), without contaminant interferences. The Q-PCR methodology was also useful to select the appropriate solid mixing time following Lacey´s mixing index (8 min). Factors and similarity principles influencing the liquid mixing process were identified in the scaling evaluation. Furthermore, the drying kinetics analysis enabled identifying a drying temperature of 35 °C, with an efficacy under controlled conditions higher HIGHLIGHTS• Technological analysis of the production for a virus-based biopesticide.• Selection of technological conditions considered to have a significant influence.• Scale-up strategy based on geometric and the dynamic similarities.• Reproducibility and repeatability assessment of the formulation process.

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