David Hidalgo1*
José Luis Ramírez2
Mercedes Navarrete1†Víctor Cevallos1†Mario Ramos1†Bill Bravo3Klever Carranza3
Víctor Montes3
Adalberto Á. Pérez de León4- 1Laboratory of Plant Protection, Instituto Nacional de Investigaciones Agropecuarias (INIAP), Santo Domingo Research Station, La Concordia, Ecuador
- 2Crop Bioprotection Research Unit, National Center for Agricultural Utilization Research, United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Peoria, IL, United States
- 3Facultad de Veterinaria, Universidad Técnica de Manabí, Portoviejo, Ecuador
- 4San Joaquin Valley Agricultural Sciences Center, USDA-ARS, Parlier, CA, United States
Ecuador is one of the countries in the world where ticks and tick-borne diseases are major constraints on cattle health and productivity. The intense use of synthetic acaricides to manage tick infestations resulted in widespread acaricide resistance in the tick Rhipicephalus microplus, which is known to infest over 75% of the farms where cattle are raised in the country. Sustainable and environmentally friendly alternatives to control the cattle tick R. microplus are needed urgently. This minireview describes a project at the Santo Domingo Experimental Station of the National Institute of Agricultural Research to develop biopesticides for tick management to exemplify advances in collaborative research on the use of entomopathogenic fungi as active ingredients, or mycopesticides, in formulations to control R. microplus. Research and development conducted in multiple phases revealed the in vitro and in vivo acaricidal properties of B. bassiana sensu lato (s.l.) strain INIAP L3B3. These efforts followed efficacy and safety norms issued by the government agency of Ecuador in charge of controls and regulations to protect an improve animal health, plant health, and food safety. Results described herein indicate that B. bassiana s.l. INIAP L3B3 can be registered as an eco-friendly mycopesticide alternative to synthetic chemical acaricides or could complement conventional chemical acaricide applications for integrated R. microplus management programs in support of sustainable cattle raising in Ecuador.
1 Introduction
Different species of entomopathogenic fungi have lethal effects on the tick Rhipicephalus microplus (López et al., 2009; Cafarchia et al., 2022), which is considered the most economically important hematophagous ectoparasite and vector of pathogens causing significant morbidity and mortality among cattle herds globally (Pérez de León et al., 2020). Like in neighboring countries of South America where this cattle tick is present (Rodríguez-Hidalgo et al., 2017), livestock farmers in Ecuador also face widespread resistance to synthetic acaricides, which are extensively used to control ticks including R. microplus (Lopez-Arias et al., 2014; Rodríguez-Hidalgo et al., 2017; Pérez-Otáñez et al., 2024c; Pérez-Otáñez et al., 2024a). Widespread resistance to synthetic acaricides highlights the need for alternative solutions, such as the development and commercialization of biopesticidal formulations utilizing entomopathogenic fungi as active ingredients, or mycopesticides (Jiang and Wang, 2023). Eco-friendly technologies like mycopesticides present a promising method for integrated R. microplus management (Angelo et al., 2022). However, there is a need to innovate mycopesticide formulations that are highly virulent, cost-effective, and stable (da Paixão et al., 2023; Friuli et al., 2023; da Silva Medina et al., 2024). Such advancements could enhance the commercialization and use of mycopesticides with other technologies to manage the parasitic and non-parasitic life stages of R. microplus infesting cattle and pastures, respectively. Relative to other countries in South America (Fernandes and Bittencourt, 2008; Moncada et al., 2015), research and development of effective and safe entomopathogenic fungi formulations commercially available for use under field conditions to control R. microplus in Ecuador remains to be fully realized.
This minireview highlights collaborative efforts by Ecuador’s National Institute of Agricultural Research (INIAP), particularly at the Santo Domingo Experimental Station (INIAP-SDES), through the project “Isolation, Maintenance, Reactivation, and Pelletizing of Entomopathogenic Fungi”. An objective of this project is to develop biopesticides based on entomopathogenic fungi to control ticks (INIAP, 2021). Previous studies in Ecuador documented the utility of entomopathogenic fungi to control R. microplus testing experimental mycopesticide formulations containing Lecanicillium lecanii, Metarhizium anisopliae, or Beauveria bassiana (Navarrete et al; Piguave, 2016; Morocho, 2019; Lopez and Masiel, 2021). However, cattle producers in Ecuador still lack a registered mycopesticide product that can be used with other technologies for integrated R. microplus management in their farms. Figure 1A provides an overview of the research and development pathway for registering and commercializing a mycopesticide product facilitated by the collection of Ecuadorian entomopathogenic fungi at INIAP (Arahana et al., 2013). This collection enabled bioassays to identify strains with acaricidal properties. Research progress with the INIAP-SDES project to deliver a registered mycopesticide product for R. microplus control is described below.
Figure 1. (A) Theoretical research and development approach to register and commercialize a mycopesticide formulation to manage the cattle tick Rhipicephalus microplus in Ecuador. *Adapted from Hidalgo (Hidalgo Mata et al., 2019). Phase 1: concept based on pre-existing collection including entomopathogenic fungi; Phase 2: includes mammalian and non-target organism safety determinations; Phase 3: includes photo and thermostability optimization, determination of synergy with other pesticides; Phase 4: adheres to federal regulatory framework of Ecuador to register and commercialize agricultural products (https://www.agrocalidad.gob.ec/?page_id=39111). (B) Phenotypic characteristics in R. microplus ticks following infection with B. bassiana s.l. INIAP L3B3, note color and size changes in fungus-infected ticks. (C) Adult tick cadaver in upper right corner, and tick egg-mass in lower left section, covered with mycelial growth.
2 Tick model selection
R. microplus is the most common tick infesting cattle in Ecuador (Pérez-Otáñez et al., 2024b). It is estimated that over 75% of the cattle farms in the country are infested with R. microplus (Vizcarra et al., 2015). In addition to direct blood feeding effects associated with its parasitic lifestyle, R. microplus also transmits pathogens like Babesia bovis, B. bigemina, and Anaplasma marginale that impact the health and diminish the productivity of cattle herds in Ecuador (Maya-Delgado et al., 2020). Acaricide resistance complicates R. microplus control based on chemical treatments with synthetic acaricides widely available to farmers (Paucar et al., 2022). Thus, the INIAP-SDES project targeted R. microplus to develop biopesticides based on entomopathogenic fungi for tick control.
3 Phase 1: selection of entomopathogenic fungal strains active against R. microplus
From the collection mentioned above, 80 fungal isolates obtained from a field survey with palm weevil (Rhynchophorus palmarum) traps were of particular interest. Bioassay of these isolates with the rice weevil (Sitophilus oryzae) yielded nine entomopathogenic strains native to the province of Santo Domingo de los Tsáchilas where the majority of the territory has a tropical climate with parts to the east in the Andean foothills with temperate climate (INIAP, 2021). B. bassiana s.l. strains INIAP L3B3 and INIAP O2 were selected for their activity against R. microplus (Figures 1B, C).The INIAP O2 strain was dropped from further testing because of lower activity against R. microplus than the L3B3 (data not shown). Fungal strains were propagated using rice as the solid substrate. However, during the initial bioassays with B. bassiana s.l. INIAP L3B3 it was observed that, despite a notable increase in conidia production, the highest mortality achieved was 8.3% on day 9 post-treatment at a concentration of 6.9 x 106 conidia/ml. Suspecting loss of virulence, the experiments were repeated using fungi recovered from tick larva cadavers that had sporulated. Following this reactivation, the median lethal dose (LD50) of B. bassiana s.l. INIAP L3B3 was 1.29 x 104 spores/ml (Herrera Montero et al., 2024). The molecular characteristics of B. bassiana s.l strain INIAP L3B3 was provisionally identified through Sanger sequencing of the ITS gene (unpublished data). Comprehensive characterization is yet to be completed.
A farm in the area of Santo Domingo de los Tsáchilas province where populations of R. microplus resistant to acaricides are known to infest cattle was selected for pilot study with B. bassiana s.l. INIAP L3B3 (Bravo et al., 2022). These experiments are described in videos available on the INIAP online channel (INIAP, 2022; INIAP, 2023). By comparison to the effects of chemical treatment by spray with a commercially available combination of 40% ethion + 10% cypermethrin using a backpack pump, R. microplus started to show the effects of entomopathogenic infection as early as 4 days after cattle infested naturally were sprayed similarly with B. bassiana s.l. INIAP L3B3 at a dose of 240 grams/10 liters of dechlorinated water (Figures 2A, B). Using the average number of R. microplus counted in the inguinal area as indicator of the infestation level in cattle in the control group, the relative effect of repeated treatment between B. bassiana s.l. INIAP L3B3 and 40% ethion + 10% cypermethrin was comparable, especially after the second reapplication (Figure 2B). These results were obtained without using the target lethal dose of 107 because by dissolving the 1.43×108 conidia/g in 10 liters of water the maximal concentration of conidia sprayed on cattle was 106.
Figure 2. (A) Drying and detachment of ticks following treatment with entomopathogenic fungal strain B. bassiana s.l. INIAP L3B3. Photos of the same bovine through time depicting reduction of tick infestations following treatment with B. bassiana s.l. INIAP L3B3 experimental formulation. (B) Temporal effect on average number of R. microplus ticks in each group of cattle sprayed with B. bassiana s.l. INIAP L3B3, commercial chemical acaricide mixture of 40% Ethion + 10% Cypermethrin, or water (Control). Dots represent the average number of ticks counted in inguinal area for each of the 3 cattle groups (n= 5) on the date shown. Red arrows indicate dates of chemical acaricide or fungal re-application. Graph generated with data from Bravo and Carranza, 2022.
Greenhouse studies revealed that B. bassiana s.l. INIAP L3B3 was also active against R. microplus eggs, thus expanding the potential use of this fungal strain (Ligña Sangucho et al., 2024). The study utilized various concentrations of conidia (ranging from 7.42x107 to 7.42x103 conidia/ml) applied to tick eggs in both sterile and non-sterile soil. The results showed an ovicidal efficacy of 20% at a concentration of 7.42x107 conidia/ml within six to ten days, with higher efficacy in non-sterile soil. The conidia remained viable for up to twenty days, effectively controlling not only eggs but also larvae and nymphs.
4 Phase 2: safety assessment of B. bassiana s.l. INIAP L3B3
B. bassiana is generally regarded as safe for mammals, non-target organisms and the environment (INIAP, 2023). Scientific literature on safety evaluations of B. bassiana strains intended for use as active ingredients in mycopesticide products was reviewed to observe best practices handling strain INIAP L3B3 in the laboratory and the pilot field study (Cazorla Perfetti et al., 2015; Authority et al., 2020; Sayed Ali et al., 2022; Ligña Sangucho et al., 2024). Previous reports indicated that other B. bassiana strains were safe to treat cattle infested with R. microplus (Navarrete et al., 2017; Lopez and Masiel, 2021).
No adverse reactions to the water formulation containing B. bassiana s.l. INIAP L3B3 were observed in the sprayed cattle. Cattle appeared normal after reapplication of the B. bassiana s.l. INIAP L3B3 spray formulation. No health concerns associated with spraying B. bassiana s.l. INIAP L3B3 were reported by the farm workers that treated the cattle.
5 Phase 3: potentiation and advanced mycopesticide formulation
Based on previous studies documenting that acaropathogenic activity of entomopathogenic fungi could be potentiated by the synergistic effect of other substances (Marrone, 2019; da Silva Medina et al., 2024), the combined effect of B. bassiana s.l. INIAP L3B3 with other organic and synthetic acaricides was tested. Bioassays evaluated the control of R. microplus combining B. bassiana s.l. INIAP L3B3 with plant extracts (eucalyptus, thyme, onion, and garlic) and the synthetic acaricide mixture containing cypermethrin + ethion (Herrera Montero et al., 2024). Compatibility test between B. bassiana and the botanical or chemical compounds were evaluated in vitro prior to testing with tick larvae. These bioassays indicated that while fresh garlic extract was acaricidal, it also inhibited conidial germination when combined with B. bassiana. Among the botanical extracts tested, only fresh garlic extract was effective at a concentration of 45 ml/l. Combining B. bassiana with garlic yielded an LD50 of 1.63 x 104 spores/ml, though garlic initially inhibited fungal germination until allicin, the active component, dissipated. The combination of B. bassiana s.l. INIAP L3B3 conidia with the chemical acaricide (cypermethrin + ethion) indicated synergy between the two (Herrera Montero et al., 2024). This resulted in exponential reduction of the LD50 for B. bassiana conidia from 1.14x 104 when used alone to a LD50 of 1.18 x 103 conidia/ml and 0.5ml/L of chemical acaricide when used in combination.
The optimization of solid-state mass production of conidia from B. bassiana s.l. INIAP L3B3 was explored to develop large-scale applications. Enhancement of eighteen formulations was evaluated with nutritional supplements (palm kernel meal, quinoa flour, brewer’s yeast, and powdered milk) combined with rice and barley (Romero Suarez et al., 2024). These experiments revealed significant differences in conidia production. The best substrate was 2.5% brewer’s yeast + 97.5% rice + Beauveria spp. yielding 6.9 x 109 conidia/g. The highest colony-forming units (CFU) concentration achieved was 2.64 x 1010 CFU/g, while purity percentages did not significantly differ among treatments. These findings were applied to scale up production of B. bassiana s.l. INIAP L3B3 conidia. Formulation improvements ensured a high-quality mycopesticide effective against R. microplus.
6 Phase 4: registration and production
Registration of mycopesticides in Ecuador is a process that requires adherence to efficacy and safety norms issued by Agrocalidad, which is the government Agency of Ecuador in charge of controls and regulations to protect an improve animal health, plant health, and food safety. The process involves an application that includes detailed information about the applicant, the producing company, the biological characterization of the fungus, and the results of biological efficacy trials conducted in the field (Agrocalidad, 2022). Efficacy data must be generated according to the “Instructions for the Approval, Execution, and Supervision of Efficacy Tests of Pesticides and Related Products for Agricultural Use in Ecuador” issued by (Agrocalidad, 2021). Per the Instructions, trials must be supervised by authorized agencies or conducted in collaboration with universities or research centers.
The INIAP-SDES project is expanding collaboration with the National Livestock and Pasture Program of Ecuador to develop and execute confirmatory tests to treat more cattle in multiple farms with a candidate formulation of B. bassiana s.l. INIAP L3B3. It is expected that these confirmatory tests will generate the efficacy and safety information needed to register a mycopesticide product in Ecuador with B. bassiana s.l. INIAP L3B3 as the active ingredient. A plan for the commercial production of mycopesticides by SDES-INIAP was presented to the national government, and the general direction of INIAP is exploring the possibility of licensing production and commercialization to private companies.
7 Discussion
The INIAP-SDES project to research and develop mycopesticides for tick control described herein addresses the problem with widespread multi-acaricide resistant populations of R. microplus in Ecuador. R. microplus and associated diseases are considered the main health burden and cause of economic loss in cattle herds nationally (Vizcarra et al., 2015; Rodríguez-Hidalgo et al., 2017; Maya-Delgado et al., 2020; Pérez-Otáñez et al., 2024c; Pérez-Otáñez et al., 2024a). Dividing the project into 4 phases depicted in Figure 1B facilitated the organization of research and development efforts, monitoring progress, and adaptation of the strategy for completion within the projected timeline. Although progress up to Phase 3 was documented, partnership with the National Livestock and Pasture Program of Ecuador is allowing the project to be completed as planned (Bustillo Pardey and Marin, 2002; INIAP, 2022; Pérez-Otáñez et al., 2024a). The loss of virulence of the B. bassiana s.l L3B3 strain jeopardized the project, requiring the experiment to be repeated after reactivating the fungus on tick larvae to recover its virulence (Herrera Montero et al., 2024). The loss of virulence in entomopathogenic fungi can be caused by factors such as successive culturing without proper reactivation, inadequate storage conditions, or genetic changes. To restore the virulence of entomopathogenic fungi, it is crucial to conduct passages on the target insect and maintain strict quality control during the reactivation process, which is essential to ensure the fungus’s effectiveness in biological control applications (Bustillo Pardey and Marin, 2002; Tomilova et al., 2023). Although acaricide resistance was not confirmed in the R. microplus infesting cattle sprayed with the experimental B. bassiana s.l. INIAP L3B3 formulation, the management of resistant cattle ticks was recommended because 83% of farms sampled in Santo Domingo de los Tsáchilas province where acaricides are used intensely were infested with R. microplus (Pérez-Otáñez et al., 2024b). Additionally, the sustained number of ticks on the control cattle sprayed with water in the pilot study suggested that pastures in the farm were infested with the non-parasitic life stages of R. microplus. Field tests are needed to confirm the ovicidal activity observed in the greenhouse studies, which suggests that B. bassiana s.l. INIAP L3B3 formulations could be used in integrated tick management to target multiple life stages of R. microplus. The ovicidal characteristic is particularly relevant, as the commercial formulation of M. brunneum F52 used in the control of Anoplophora glabripennis showed an ovicidal effect of up to 40% under specific high-humidity conditions in the field (Goble et al., 2015).
Field tests are needed to confirm the synergistic effect observed in vitro when B. bassiana s.l. INIAP L3B3 was combined with the cypermethrin + ethion acaricidal mixture. This highlights the nuances of formulating acaricide mixtures with entomopathogenic fungi on acaricidal efficacy because the compatibility of B. bassiana can vary depending on the class of acaricide or members of the same acaricide class used in the mix (Cafarchia et al., 2022; Friuli et al., 2023). Our observations confirmed the reported acaricidal property of garlic and despite inhibiting fungal germination, B. bassiana s.l. INIAP L3B3 could be alternated with garlic extract to treat infested pastures (Herrera Montero et al., 2024). Advances in formulation science and technology could allow further improvements for efficient mass production of B. bassiana s.l. INIAP L3B3. The project is considering a pellet formulation using local inputs to make production of a high-quality B. bassiana s.l. INIAP L3B3-based mycopesticide more cost-effective (Hidalgo and Adrian, 2019; Hidalgo Mata and Tello Torres, 2022).
Development of a robust regulatory framework to register and control mycopesticides and other biopesticides in Ecuador since the second decade of this century provided a critical path for this project (Agrocalidad, 2021; Agrocalidad, 2022). Clarity on the registration process expedites implementation research and the translation of efficacy and safety results from laboratory and field studies (Jiang and Wang, 2023), which could accelerate the commercialization of mycopesticides for sustainable R. microplus control. The results obtained with this project thus far indicate that B. bassiana s.l. INIAP L3B3 can be registered as an eco-friendly mycopesticide alternative to synthetic chemical acaricides or could complement conventional chemical acaricide applications for integrated R. microplus management programs in support of sustainable cattle raising in Ecuador.
Author contributions
DH: Writing – original draft, Writing – review & editing, Conceptualization, Funding acquisition, Investigation. JLR: Writing – original draft, Writing – review & editing. MN: Conceptualization, Project administration, Resources, Writing – review & editing. VC: Project administration, Resources, Writing – review & editing. MR: Project administration, Validation, Writing – review & editing. BB: Formal Analysis, Investigation, Methodology, Writing – review & editing. KC: Formal Analysis, Investigation, Methodology, Writing – review & editing. VM: Formal Analysis, Methodology, Writing – review & editing. AAPL: Writing – original draft, Writing – review & editing.
Funding
The author(s) declare that financial support was received for the research and/or publication of this article. Research by DH was funded by INIAP-SDES project “Isolation, Maintenance, Reactivation, and Pelletizing of Entomopathogenic Fungi”. USDA-ARS appropriated project 5010-22410-022-000-D is the primary source of research funding for JLR. Research by AAPL is funded in part by USDA-ARS appropriated project 2034-22000-015-000-D.
Acknowledgments
The authors are grateful for the support of cattle producers and agricultural research stakeholders to innovate sustainable technologies for integrated tick management.
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Publisher’s note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
Author disclaimer
Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. The mention of a proprietary product does not constitute an endorsement or a recommendation by the USDA for its use. USDA is an equal opportunity provider and employer.
References
Agrocalidad (2021). Instructivo para la aprobación, ejecución y supervisión de ensayos de eficacia de plaguicidas y productos afines de uso agrícola en Ecuador.
Agrocalidad (2022). Manual Técnico de Procedimientos para el registro y control de agentes de control Biológico, extractos vegetales, preparados minerales y semioquímicos.
Angelo I. C., Ramirez J. L., Golo P. S., Fernandes É. K. K., Pedrini N., Bittencourt V. R. E. P. (2022). Editorial: entomopathogenic fungi for the control of arthropod pests. Front. Physiol. 13. doi: 10.3389/fphys.2022.885131
Arahana V., Bastidas N., Torres M., Gonzalez P. (2013). Genetic diversity of an entomopathogenic fungi collectionfrom Ecuador using a modifiedAFLP approach. Avances en Cienc. e Ingenierias 5, B49–B57.
Authority E. F. S., Anastassiadou M., Arena M., Auteri D., Brancato A., Bura L., et al. (2020). Peer review of the pesticide risk assessment of the active substance Beauveria bassiana strain 203. EFSA J. 18, e06295. doi: 10.2903/j.efsa.2020.6295
Bravo V., Bill K., Carranza M., Klever A. (2022). Uso de hongos entomopatogenos como alternativa en el control biologico de garrapatas en la provincia de Santo Domingo de los Tsachilas (Santo Domingo, EC: INIAP-Estación Experimental Santo Domingo, Protección Vegtal, 2022: Universidad Tecnica de Manabi).
Bustillo Pardey A. E., Marin M. P. (2002). ¿Cómo reactivar la virulencia de Beauveria bassiana para el control de la broca del café? (Turrialba (Costa Rica: CATIE).
Cafarchia C., Pellegrino R., Romano V., Friuli M., Demitri C., Pombi M., et al. (2022). Delivery and effectiveness of entomopathogenic fungi for mosquito and tick control: Current knowledge and research challenges. Acta Tropica. 234, 106627. doi: 10.1016/j.actatropica.2022.106627
Cazorla Perfetti D., Acosta Quintero M., Morales Moreno P. (2015). Evaluación de la Patogenicidad Inhalatoria del Micoinsecticida Beauveria bassiana LF14 en Ratones. Rev. Investigaciones Veterinarias del Perú. 26, 565–576. doi: 10.15381/rivep.v26i4.11213
da Paixão F. R. S., Muniz E. R., Catão A. M. L., Santos T. R., Luz C., Marreto R. N., et al. (2023). Microsclerotial pellets of Metarhizium spp.: thermotolerance and bioefficacy against the cattle tick. Applied Microbiology and Biotechnology 107, 2263–2275. doi: 10.1007/s00253-023-12467-7
da Silva Medina G., Rotondo R., Rodríguez G. R. (2024). Innovations in agricultural bio-inputs: commercial products developed in Argentina and Brazil. Sustainability. 16, 2763. doi: 10.3390/su16072763
Fernandes É. K. K., Bittencourt V. R. E. P. (2008). Entomopathogenic fungi against South American tick species. Exp. Appl. Acarology. 46, 71–93. doi: 10.1007/s10493-008-9161-y
Friuli M., Pellegrino R., Lamanna L., Nitti P., Madaghiele M., Demitri C. (2023). Materials engineering to help pest control: A narrative overview of biopolymer-based entomopathogenic fungi formulations. J. Fungi. 9, 918. doi: 10.3390/jof9090918
Goble T. A., Hajek A. E., Jackson M. A., Gardescu S. (2015). Microsclerotia of metarhizium brunneum F52 applied in hydromulch for control of asian longhorned beetles (Coleoptera: cerambycidae). J. Economic Entomology. 108, 433–443. doi: 10.1093/jee/tov013
Herrera Montero L. M., Romero Peláez A. B., Gómez Mendoza G. A., Navarrete Párraga M. E., David Adrian H. M. (2024). Control de garrapatas mediante Beauveria bassiana cepa INIAP L3B3 en combinación con moléculas orgánicas y químicas. Emergentes - Rev. Científica. 4, 583–596. doi: 10.60112/erc.v4i2.166
Hidalgo M., Adrian D. (2019). Desarrollo de una metodología de peletización para Beauveria bassiana con insumos Peruanos (Biblioteca Agricola Nacional, Repositorio Institucional, Universidad Nacional Agraria La Molina: Universidad Nacional Agraria La Molina).
Hidalgo Mata D. A., Tello Torres C. M. (2022). Manual para la producción de hongos entomopatógenos y análisis de calidad de bioformulados.
INIAP (2021). Informe anual Instituto Nacional de Investigaciones Agropecuarias (INIAP) (Estación Experimental Santo Domingo). Available at: http://repositorio.iniap.gob.ec/handle/41000/5622. (Accessed November 10, 2024)
INIAP (2022). Development of Fungal-based biopesticides. Available online at: https://www.youtube.com/watch?v=OcUudq7ZnwY&t=8s. (Accessed August 5, 2024)
INIAP (2023). Tick detachment post treatment with entomopathogenic fungi formulations. Available online at: https://www.youtube.com/watch?v=l3263Plef9k. (Accessed August 5, 2024)
Jiang Y., Wang J. (2023). The registration situation and use of mycopesticides in the world. J. Fungi. 9, 940. doi: 10.3390/jof9090940
Ligña Sangucho K. D., Gómez Mendoza G. A., Ávila Vaca R. R., Navarrete Párraga M. E., Hidalgo Mata D. A. (2024). Efecto ovicida de Beauveria spp. sobre huevos de garrapata Rhipicephalus microplus en invernadero. Emergentes - Rev. Científica. 4, 597–610. doi: 10.60112/erc.v4i2.167
López E., López G., Orduz S. (2009). Control de la garrapata Boophilus microplus con Metarhizium anisopliae, estudios de laboratorio y campo. Rev. Colombiana Entomología. 35, 42–46. doi: 10.25100/socolen.v35i1.9187
Lopez S., Masiel L. (2021). Bioensayo para el control de la garrapata (Rhipicephalus microplus) de bovino utilizando diluciones del hongo Beauveria bassiana, manta (Universidad Laica “Eloy Alfaro” de Manabi).
Lopez-Arias A., Villar-Argaiz D., Chaparro-Gutierrez J. J., Miller R. J., De Leon A. A. P. (2014). Reduced Efficacy of Commercial Acaricides against Populations of Resistant Cattle Tick Rhipicephalus microplus from Two Municipalities of Antioquia, Colombia. Environ. Health Insights 8s2, EHI.S16006. doi: 10.4137/EHI.S16006
Marrone P. G. (2019). Pesticidal natural products – status and future potential. Pest Manage. Science. 75, 2325–2340. doi: 10.1002/ps.v75.9
Maya-Delgado A., Madder M., Benítez-Ortíz W., Saegerman C., Berkvens D., Ron-Garrido L. (2020). Molecular screening of cattle ticks, tick-borne pathogens and amitraz resistance in ticks of Santo Domingo de los Tsáchilas province in Ecuador. Ticks Tick-borne Diseases. 11, 101492. doi: 10.1016/j.ttbdis.2020.101492
Moncada G. A., Villar A. D., Chaparro G. J., Angulo A. J., Mahecha L. L. (2015). Aproximación al uso de hongos entomopatógenos y vacunas para el control sostenible de garrapatas en sistemas ganaderos: revisión. Avances en Investigación Agropecuaria 19.
Morocho S. (2019). Evaluación in vivo de Metarhizium anisopliae (Metchnikoff) y Beauveria bassiana (Balls) sobre garrapatas (Acari: Ixodidae) que parasitan el ganado vacuno (en El Chaco, Ecuador: Universidad de las Fuerzas Armadas). DV.
Navarrete F., Intriago A., Álava N., Aguilar N. (2017). Determinacion de la dosis de los hongos entomopatógenos (Beauveria bassiana y Lecanicillium lecanii) para el control de garrapata, en ganado vacuno in vitro e in vivo2017. Available online at: https://www.engormix.com/ganaderia/garrapatas-bovinos/determinacion-dosis-hongos-entomopatogenos_a41154/. (Accessed August 5, 2024)
Paucar V., Pérez-Otáñez X., Rodríguez-Hidalgo R., Perez C., Cepeda-Bastidas D., Grijalva J., et al. (2022). The associated decision and management factors on cattle tick level of infestation in two tropical areas of Ecuador. Pathogens. 11, 403. doi: 10.3390/pathogens11040403
Pérez de León A. A., Mitchell Rd I. I. I., Watson D. W. (2020). Ectoparasites of cattle. Veterinary Clinics: Food Anim. Practice. 36, 173–185. doi: 10.1016/j.cvfa.2019.12.004
Pérez-Otáñez X., Paucar-Quishpe V., Saegerman C., Grijalva-Olmedo J., Pérez-Escalante C., Jácome L., et al. (2024a). Drivers and evolution of acaricide resistance and multi-resistance in two Ecuador’s subtropical livestock farming areas. Veterinaria Italiana. doi: 10.12834/VetIt.3471.23969.2
Pérez-Otáñez X., Rodríguez-Hidalgo R., Enríquez S., Celi-Erazo M., Benítez W., Saegerman C., et al. (2024b). High-resolution prediction models for Rhipicephalus microplus and Amblyomma cajennense s.l ticks affecting cattle and their spatial distribution in continental Ecuador using bioclimatic factors. Exp. Appl. Acarology 92, 439–462. doi: 10.1007/s10493-023-00883-3
Pérez-Otáñez X., Vanwambeke S. O., Orozco-Alvarez G., Arciniegas-Ortega S., Ron-Garrido L., Rodríguez-Hidalgo R. (2024c). Widespread acaricide resistance and multi-resistance in Rhipicephalus microplus in Ecuador and associated environmental and management risk factors. Ticks Tick-borne Diseases. 15, 102274. doi: 10.1016/j.ttbdis.2023.102274
Piguave R. (2016). Evaluación del efecto acaricida del hongo entomopatógeno (Lecanicillium lecanii) en el control de las garrapatas en el ganado vacuno (Universidad de Guayaquil; Facultad de Ciencias para el Desarrollo).
Rodríguez-Hidalgo R., Pérez-Otáñez X., Garcés-Carrera S., Vanwambeke S. O., Madder M., Benítez-Ortiz W. (2017). The current status of resistance to alpha-cypermethrin, ivermectin, and amitraz of the cattle tick (Rhipicephalus microplus) in Ecuador. PLoS One 12, e0174652. doi: 10.1371/journal.pone.0174652
Romero Suarez J. M., Santos Pazmiño J. P., Gómez Mendoza G. A., Navarrete Párraga M. E., David Adrián H. M. (2024). Optimización de Sustratos Sólidos para la Producción de Conidios de Beauveria spp. e Isaria spp. Emergentes - Rev. Científica. 4, 611–626. doi: 10.60112/erc.v4i2.168
Sayed Ali S., El-Saadany H. M., Kotb G. A. M., Elshaer N., Melebary S. J., Soliman S. M., et al. (2022). Biosafety evaluation of two Beauveria bassiana products on female albino rats using acute oral test. Saudi J. Biol. Sci. 29, 103293. doi: 10.1016/j.sjbs.2022.103293
Tomilova O. G., Kryukov V. Y., Kryukova N. A., Tolokonnikova K. P., Tokarev Y. S., Rumiantseva A. S., et al. (2023). Effects of passages through an insect or a plant on virulence and physiological properties of the fungus Metarhizium robertsii. PeerJ. 11, e15726. doi: 10.7717/peerj.15726
Keywords: Beauveria bassiana, mycopesticide, cattle tick, Rhipicephalus microplus, microbial control
Citation: Hidalgo D, Ramírez JL, Navarrete M, Cevallos V, Ramos M, Bravo B, Carranza K, Montes V and Pérez de León AÁ (2025) Research advances in Ecuador on use of entomopathogenic fungi for control of the cattle tick, Rhipicephalus microplus: the case of Beauveria bassiana sensu lato strain INIAP L3B3. Front. Fungal Biol. 6:1492395. doi: 10.3389/ffunb.2025.1492395
Received: 06 September 2024; Accepted: 06 March 2025;
Published: 03 April 2025.
Edited by:
Chengshu Wang, Chinese Academy of Sciences (CAS), ChinaReviewed by:
Mavis Agyeiwaa Acheampong, University of Ghana, GhanaCopyright © 2025 Hidalgo, Ramírez, Navarrete, Cevallos, Ramos, Bravo, Carranza, Montes and Pérez de León. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: David Hidalgo, ZGF2aWQuaGlkYWxnb0BpbmlhcC5nb2IuZWM=; ZGFhZGhpbWFAZ21haWwuY29t
†ORCID: Mercedes Navarrete, orcid.org/0000-0001-7862-0697, Victor Cevallos, orcid.org/0000-0003-4390-8193, Mario Ramos, orcid.org/0000-0003-2264-2808