Horizontal transmission of Beauveria bassiana spores using infected males and inoculation device: impact on survival and fecundity of Ceratitis capitata (Diptera: Tephritidae)

Horizontal transmission of Beauveria bassiana spores using infected males and inoculation device: impact on survival and fecundity of Ceratitis capitata (Diptera: Tephritidae) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Horizontal transmission of Beauveria bassiana spores using infected males and inoculation device: impact on survival and fecundity of Ceratitis capitata (Diptera: Tephritidae) Cesar Galvez, Salvador Flores, Sergio Campos, Francisco Ramirez y Ramirez, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2180398/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Feb, 2023 Read the published version in Phytoparasitica → Version 1 posted 7 You are reading this latest preprint version Abstract The mode of transmission of fungus spores (horizontal transmission or assisted auto-dissemination) directly influences the effectiveness of a fungal pathogen when used as a control agent. Fungal infections cause physiological alterations leading to the host's death. During this process, the fungus uses the energy reserves in the hemolymph of insects, affecting the development and performance of individuals and, therefore, the demographic features of their populations. In this work, we evaluated topical inoculation and an auto-disseminator device in the transmission of Beauveria bassiana (Balsamo) Vuillemin conidia to Ceratitis capitata Wiedemann (Diptera: Tephritidae). Survival and fecundity were negatively affected by the action of the fungus, and mortality was influenced by the inoculation method. Inoculated sterile males were as competitive as untreated males and reduced the fecundity and survival of females. We conclude that the pathogenic action of B. bassiana reduces the survival and fecundity parameters of C. capitata infected by horizontal transmission, while the behavioral response of treated sterile males is similar to that of sterile-fertile untreated males. We discuss the potential use of this strategy as part of the pest management of C. capitata . Beauveria bassiana Ceratitis capitata entomopathogenic fungus sterile insect technique integrated pest management. Figures Figure 1 Figure 2 Introduction In natural ecosystems, entomopathogenic fungi (EPF) play an important role in the regulation of insect populations (Goettel et al., 2010 ) and therefore have great potential as biocontrol agents (Dimbi et al., 2003 ). Infection by EPF begins with the adherence of the conidium and penetration through the cuticle, followed by the consumption of nutrients present in the insect’s hemolymph by the fungus. During mycelial growth, toxins are produced as secondary metabolites, which may cause physiological and behavioral alterations (Xia et al., 2002 ; Khachatourians & Qazi, 2008 ; Molnár et al., 2010 ; Anderson et al., 2011 ; Jin et al., 2015 ). The fecundity of infected hosts can be strongly affected during the invasion process, and thus may represent an additional component in the suppression of pest populations (Shoukat et al., 2020 ; Mkiga et al., 2020 ). Baverstock et al. ( 2010 ) mention that fungal spore transmission may occur by horizontal transmission among individuals of the same species or by assisted auto-dissemination (using artificial autoinoculation devices). Both modes of transmission have been able to cause fungal infections in some target fruit fly pests (e.g., Quesada-Moraga et al., 2008 ; Dimbi et al., 2013 ; Sookar et al., 2014 ; Patt et al., 2015 ; Toledo et al., 2017 ; Yousef et al., 2018 ). In both cases, the horizontal transmission rate has been the key factor determining the spread of the pathogen within host populations (Steinkraus, 2006 ; Baverstock et al., 2010 ). Beauveria bassiana (Balsamo) Vuillemin is a facultative fungus with a wide host range found in both temperate and tropical zones (Shah & Pell, 2003 ; Zimmermann, 2007 ). The pathogenicity of B. bassiana has been evaluated in the fruit fly genus Anastrepha spp. (De la Rosa et al., 2002 ; Toledo et al., 2007 ) and in Ceratitis capitata Wiedemann (Diptera: Tephritidae) (Ekesi et al., 2002 ; Dimbi et al., 2003 ; Toledo et al., 2006 ; Flores et al., 2013 ; Toledo et al., 2017 ). Dias et al. ( 2022a ) note that B. bassiana is one of the most promising and versatile fungus species that can function as a biocontrol agent. Toledo et al. ( 2017 ) and Montoya et al. ( 2020 ) evaluated this species with the concurrent use of autoinoculation devices and the Sterile Insect Technique (SIT), observing an increase in infected wild fruit flies, and thus proposed an additive or synergistic effect of both technologies. The Mediterranean fruit fly, C. capitata , is a highly polyphagous, multivoltine species that infects more than 353 host fruit species around the world (Papadopoulos, 2008 ). Currently, this pest has an extensive distribution in tropical and temperate zones, limiting the growth and development of international fresh fruit trade (Stewart & Johanson, 1999 ; Vera et al., 2002 ; Malacrida et al., 2007 ; Diamantidis et al., 2011 ; Dias et al., 2022b ). In Mexico, the SIT is applied to eradicate C. capitata transient entries using an integrated pest management (IPM) framework. The sterile insects are provided by the mass-rearing facility located in Metapa, Chiapas, where the insects have been inoculated with B. bassiana conidia to infect wild flies in the border with Guatemala. The use of autoinoculation devices to disseminate EPF conidia has been another strategy employed to control pest populations (SENASICA-SAGARPA, 2017; Flores et al., 2013 ; Toledo et al., 2017 ). The selected EPF strains must allow enough time for interactions to occur and conidia to disperse among conspecifics before the insect’s death. However, how the infection process affects the sexual competitiveness of males, as well as the resulting fitness of the female conidia recipients, is still not well known. Thus, the objectives of the present work were to determine the effect of the horizontal transmission of B. bassiana on the survival, fecundity, and fertility of C. capitata individuals using inoculated adults and auto-inoculation devices. We also determined the impact of the fungus infection on sterile male performance as vectors of EPF. The above will contribute to a better understanding of pathogen-host interactions, and thus reinforce these ecologically oriented control strategies within IPM. Materials And Methods Biological material. Males (sterile, fertile) and fertile females of C. capitata were provided by the Moscamed facility (SADER-SENASICA) located in Metapa, Chiapas, Mexico. After emergence, each sex was kept separately in Plexiglas cages (30 × 30 × 30 cm) with water and food (a 3: 1 mixture of sugar: hydrolyzed protein [MP Biomedicals, LLC, Santa Ana, California, USA]) until they reached sexual maturity. Laboratory conditions were maintained at 25 ± 1°C, 60 ± 10% R.H, and a photoperiod of 12:12 h light: dark. All tests were carried out inside a cage 3 m in diameter × 2 m high made of saran screen (20 by 20 mesh) in an isolated room inside the Moscamed facility to reinforce biological safety measures. The B. bassiana strain was formulated with 2 × 10 9 conidia per g with Celite 400 as inert material (Laboratorio de Organismos Benéficos, Hongos, Insectos y Nematodos, Talisman, Chiapas, Mexico). The viability of the fungus was determined before each test according to the microculture technique (Jiménez, 1992 ) and with observations of germinated conidia at 40× using an optical microscope (Dialux 20 EB; Leitz, Wetzlar, Germany). The conidia were considered viable when the length of the germ tube was greater than the diameter. The evaluations were carried out with batches of conidia with a recorded viability of ≥ 90%. Inoculation methods . Adults of C. capitata were infected with B. bassiana conidia either topically or using the inoculation device method: 1) Topical inoculation. We applied 0.05 g of the B. bassiana formulation to 50 fertile or sterile male or female C. capitata in a 50 ml Falcon conical centrifuge tube (29 mm diameter ⋅ 115 mm height), which was shaken gently for 10s for a homogeneous inoculation of all individuals. 2) Inoculation with PVC device. The device was constructed using a PVC tube (9 cm in diameter ⋅ 10 cm high) with two perforations of 1 cm in diameter in the lower part, and 100 ⋅ 10 mm Petri dish bottoms were used as lids on both ends. The interior of the tube was lined with yellow plush fabric (felt), similarly to Toledo et al. ( 2017 ), which was homogeneously impregnated with 2 g of the conidia formulation. Groups of 50 females or males were placed inside test tubes (25 ⋅ 150 mm) and then introduced through the perforations on the lower part of the PVC device. The insects were recovered after 10 min, which was enough time to ensure contact with the fungal conidia. Inoculated adults were separated by sex and kept in Plexiglas cages (30 ⋅ 30 ⋅ 30 cm) for 20 min to allow them to remove excess conidia by grooming. The effect of B. bassiana conidia on the fecundity, fertility, and survival of C. capitata adults was determined through the following tests: a) Effect of inoculation method. The effect of B. bassiana on C. capitata was determined by direct inoculation of females or by horizontal transmission through inoculated males. The combinations evaluated were: inoculated males + inoculated females, inoculated males + non-inoculated females, non-inoculated males + inoculated females. Fungal conidia were applied to adults either by topical inoculation or with the PVC device. A treatment with non-inoculated males + non-inoculated females was included as a control treatment. This resulted in seven treatments, three combinations × two inoculation methods + control. For each treatment, 10 fertile fly pairs were placed in plastic containers of 1.75 L (17.3 cm high, 21.2 × 11.3 cm) provided with water and food (3:1 mixture of sugar: hydrolyzed protein). The container was placed in a horizontal position and kept under laboratory conditions for 10 days. As oviposition substrate, the opening of the plastic container was covered with a piece of white lycra fabric (Dorian Gray Likra Pantyhose). Laid eggs fell into a petri dish with tap water placed below the container opening. Each container was one experimental unit, and each repetition was carried out with a different production batch of C. capitata. We performed five replicates per treatment for horizontal transmission. b) Inoculation of sterile males. The effect of infected sterile males on the survival, fecundity, and fertility of fertile females was compared with that of inoculated fertile males and the interaction between fertile and inoculated sterile males. The treatments were (1) inoculated sterile males and (2) non-inoculated sterile males, both interacting with fertile males. The topical method was used to apply the conidia to both sterile and fertile males. According to each treatment, 10 males and 10 females were placed in plastic containers of 1.75 L as described above. In the fertile-sterile male treatments, 5 sterile males (inoculated or healthy) and 5 fertile males were placed with 10 fertile females. Each container was an experimental unit with six replicates per treatment, each one with a different production batch of C. capitata. Fecundity, fertility, mortality, and mycosis were estimated as follows: Fecundity and egg fertility. Eggs were collected in a Petri dish bottom (150 ⋅ 20 mm) with 200 ml of sterile water placed under the oviposition substrate. For each treatment, eggs were recovered daily using a piece of black organza cloth (10 ⋅ 10 cm) and were aligned and counted using a stereomicroscope (Nikon, SMZ645, USA). A sample of 100 eggs was placed in an incubation chamber (plastic Petri dish of 100 ⋅ 10 mm, where a piece of black cloth was placed on a piece of 2-mm-thick moistened filter paper). After four days, the hatched eggs were counted using a stereomicroscope with a 4⋅ objective. Egg fertility was expressed as percentage of hatched eggs. Mortality and fungal sporulation. The number of dead adults in each treatment was recorded daily. Dead adults were disinfected for 20 sec in a 1% sodium hypochlorite solution and then washed three times with sterile distilled water to eliminate saprophytic microorganisms. Subsequently, the specimens were placed in a humid chamber (sterile wet filter paper in 100 × 10 mm plastic Petri dishes) sealed with Parafilm (Bemis Company, Inc. Neenah, WI, 54956) and kept under laboratory conditions. After five days, the flies were examined under a stereomicroscope to verify the presence of mycosis. Statistical analysis . In each test, survival curves over the course of the ten days of evaluation were compared using a log-rank test. Fecundity, number of eggs per live female per day, was log-transformed to obtain homoscedasticity prior to performing an analysis of variance (ANOVA), and mean comparisons were performed with a Tukey test (α = 0.05). Fertility, expressed as the proportion of hatched eggs, was analyzed with a GLM with binomial distribution and a logit link function. Treatment means were compared by contrasts with a significance level of 95%. In the case of horizontal transmission of B. bassiana calculated in treatments with inoculated and non-inoculated adults, mycosis in recipient flies was analyzed using a generalized linear model (GLM) with binomial distribution and a logit link function, and treatments were compared by orthogonal contrasts (α = 0.05). All analyses were conducted in JMP® 11.0.0 (JMP® SAS Institute, http://www.jmp.com ). Results a) Inoculation method Survival of females (χ 2 = 217.98, d.f. = 6, P < 0.001) and males (χ 2 = 189.34, d.f. = 6, P < 0.001) differed significantly between treatments. The survival of directly inoculated females was reduced to at least 20% at 5 days. In treatments with inoculated males, the survival of non-inoculated females showed a gradual reduction throughout the 10 days. In the control treatment, the mortality was lower than 20% after 10 days. The survival curve of females did not differ between topical inoculation and the use of an inoculation device (Fig. 1 ). The number of eggs per female was higher at day 3 and 4 in each treatment, with the control treatment having the highest number of eggs (Fig. 1 ). Fecundity was significantly affected by treatment (F = 18.58, d.f. = 6, 24, P < 0.001). The control treatment had the highest fecundity, and the treatments with directly inoculated females showed the lowest fecundity. When comparing the inoculation methods, the treatments with topical inoculation showed lower fecundity than those with the device method (Table 1 ). Table 1 Fecundity and egg fertility of Ceratitis capitata females in treatments using inoculated insects with Beauveria bassiana conidia, applied by topical or device method. Treated Sex Method of inoculation Fecundity (Eggs/♀ ± S. E.) Egg Fertility (% Eclosion ± S.E.) Transmission (% mycosis ± S.E.) ♂-♀ Device 64.3 ± 12.0 d 74.7 ± 2.1 c --- Topical 50.1 ± 10.0 d 68.6 ± 2.6 d --- ♂ Device 155.8 ± 34.6 b 78.6 ± 1.4 b 66.0 ± 12.9 a Topical 148.8 ± 34.1 bc 70.8 ± 1.6 c 74.5 ± 11.8 a ♀ Device 72.6 ± 9.9 cd 78.6 ± 2.0 b 76.0 ± 11.8 a Topical 50.0 ± 9.3 d 77.6 ± 1.5 b 80.0 ± 11.1 a None Control 243.1 ± 29.8 a 86.2 ± 1.0 a --- Means followed by the same letter in each column are not significantly different (α = 0.05) The fertility of C. capitata was significantly affected in treatments with the fungus spore (χ 2 = 399.59, d.f. = 6, P < 0.001), with the control having the highest proportion of hatched eggs. The treatments with directly inoculated males showed the lowest egg hatching percentage. The treatments with topical inoculation of males showed the lowest fertility values (Table 1 ). Fungal sporulation in inoculated adults varied from 94 to 98% between males and females in the different treatments (χ 2 = 8.23, d.f. = 7, P = 0.312). The horizontal transmission of B. bassiana did not differ significantly between treatments where inoculated and non-inoculated individuals coexisted (χ 2 = 3.76, d.f. = 3, P = 0.288) (Table 1 ). b) Infection of sterile males Female survival (χ 2 = 217.59, df = 5, P < 0.001) was significantly different between treatments. The survival curve of females in the treatments with inoculated males was significantly reduced compared to treatments with non-inoculated males (χ 2 = 216.88, d.f. = 1, P < 0.001). The survival curve of females exposed to inoculated males (χ 2 = 0.47, d.f. = 2, P = 0.789) and non-inoculated males (χ 2 = 1.15, d.f. = 2, P = 0.64) was not affected by the male strain used (Fig. 2 ). The number of eggs per female per day showed the highest peak between days 3 and 4 in each treatment. In the treatments with non-inoculated males, the number of eggs per female exhibited a gradual decrease, while there was greater variation after day 6 in the treatments with inoculated males (Fig. 1 ). The total fecundity for the entire period was significantly lower in treatments with inoculated males than in treatments with non-inoculated males (F = 17.25, d.f. = 1.25, P < 0.001). Fecundity did not differ significantly between the treatments with sterile, fertile, or sterile and fertile males (F = 0.82, d.f. = 2, 25, P = 0.450) (Table 2 ). Table 2 Fecundity and egg fertility of Ceratitis capitata females in field cage tests, using Beauveria bassiana inoculated sterile and no-inoculated fertile males. Male strain Treatment Fecundity (Eggs/♀ ± S.E.) Egg Fertility (% Eclosion ± S.E.) Transmission (% mycosis ± S.E.) Fertile No-inoculated 226.0 ± 24.7 ab 85.6 ± 0.9 a --- Inoculated 148.1 ± 40.4 ab 72.9 ± 1.5 c 73.3 ± 11.1 a Sterile-Fertile No inoculated 232.9 ± 15.8 a 64.8 ± 1.3 d --- Inoculated 150.6 ± 30.8 ab 81.3 ± 1.2 b 73.3 ± 11.1 a Sterile No inoculated 211.0 ± 29.3 ab --- --- Inoculated 125.5 ± 39.4 b --- 73.3 ± 11.1 a Means followed by the same letter in each column are not significantly different (α = 0.05). The egg fertility of C. capitata was significantly affected by treatment (χ 2 = 724.7, d.f. = 3, P < 0.001). Inoculation of fertile males significantly reduced the percentage of hatched eggs. When inoculated or non-inoculated sterile males interacted with fertile males and females, egg fertility was lower than with non-inoculated sterile males (Table 2 ). Horizontal transmission to females did not differ significantly between the three treatments that included males inoculated with B. bassiana , with a mycosis percentage of 73.3% in each treatment (Table 2 ). Discussion As expected, the survival and fecundity of C. capitata were affected by the infection of B. bassiana . The use of inoculated sterile males or autoinoculation devices resulted in the horizontal transmission of B. bassiana to untreated insects. Both inoculation methods allowed the successful interaction between treated and untreated insects, allowing thus a horizontal transmission between them, which provides a better understanding of how infections by fungi, such as B. bassiana , affect the survival and reproduction of contaminated insects. This undoubtedly will lead to the implementation of more effective and timelier IPM strategies (Shoukat et al., 2020 ) for the control of fruit fly populations. Our results support the proposal to simultaneously use the SIT and EPF as in Montoya et al. ( 2020 ). Furthermore, devices for disseminating EPF conidia are inexpensive, easy to apply, and effective (Toledo et al., 2017 ; Yousef et al., 2018 ). Our results showed a range of 65–80% horizontal transmission of B. bassiana conidia to untreated C. capitata adults. These results agree with Chergui et al. ( 2020 ), who recorded a maximum of 51.67 and 73.33% mortality on the 15th day of a contact bioassay in males and females, respectively. The success of EPF as a control agent depends mainly on the possibility of transmitting the infection among conspecifics, as well as on the behavioral response of the insect species during the host-pathogen interaction (Baverstock et al., 2010 ; Dimbi et al., 2013 ). In the case of fruit flies, fungal transmission is enhanced by the mating behavior displayed by the adults. Fruit fly males form leks (i.e., groups of males calling for matings [Shelly, 2018 ; Thaochan & Ngampongsai, 2018 ]) and compete with each other to copulate with the females present. This behavior generates several interactions that increase the chances of successful fungal transmission among conspecifics. We observed that the horizontal transmission of conidia was higher with the topical method than with the auto-inoculation device, which could be due to the topical method resulting in a larger number of conidia per fly than the device method. Chergui et al. ( 2020 ) indicate that different methods of B. bassiana application produce different results. According to Kaneshiro et al. ( 1993 ), a larger amount of conidia could generate female rejection of sterile males, although the excess of EPF could be reduced by grooming (Baverstock et al., 2010 ). Moreover, grooming allows conidia to be deposited in preferred gathering sites for adults, for example, during lek formation (Arita & Kaneshiro, 1985 , 1989 ; Thaochan & Ngampongsai, 2018 ). The survival of treated adults was reduced from 7 to 10 days. During this period, females can oviposit before dying; however, their reproductive potential was highly reduced by the EPF infection (see Toledo et al., 2007 ). The effect of B. bassiana of reducing the fecundity, fertility, and survival of C. capitata females in this study was due to the physiological alterations derived from the pathogenic infection (Jin et al., 2015 ; Usman et al., 2021 ). For example, the depletion of sugar and other compounds in the insect hemolymph by EPF (e.g., Xia et al., 2002 ; Jin et al., 2015 ; Peng et al., 2015 ) has serious consequences on the fitness parameters of the host insects (Jin et al., 2015 ). The observed reduction in the oviposition rate and fertility of infected females has been highly associated with the action of EPF, which negatively impacts insect populations as a resulting compensation for their delayed mortality (Dimbi et al., 2013 ). The adverse effects on fecundity can also be caused by the antifeedant activity of B. bassiana during the invasive process (Ekesi, 2001 ). Our results showed that fecundity was lower in directly infected females than in females infected by inoculated fertile males. We also observed a negative effect on egg hatching when fertile males were inoculated than when inoculated sterile males interacted with non-inoculated fertile males. The use of insects as vectors of biocontrol agents has been proposed as a strategy to improve biological control programs through the horizontal transmission of pathogens (Vickers et al., 2004 ; Llacer et al., 2013; Diouf et al., 2022 ). The application of this strategy in fruit fly programs allows the release of inoculated sterile males to disseminate conidia into wild populations of C. capitata (Toledo et al., 2017 ). Recently, Diouf et al. ( 2022 ) introduced the term “boosted SIT” to refer to the use of sterile insects as vectors of biocides to trigger an epizootic in wild populations. For successful results, quality parameters, such as dispersion and competitiveness, of sterile males must remain unaffected after inoculation (Novelo-Rincon et al., 2009; Ramírez y Ramírez et al., 2020), at least long enough to interact with a wild population. In conclusion, the inoculation of B. bassiana conidia using a device or the topical method was effective in causing mycosis in adult C. capitata , and the horizontal transmission of fungal spores significantly reduced the fecundity, egg fertility, and survival of C. capitata females. These results support the proposal of including this strategy in IPM programs for the suppression of Mediterranean fruit fly populations. Declarations Ethics approval The research meets ethical guidelines and adheres to the legal requirements of the study country. This research does not involve human subjects. Consent to participate Not applicable. Consent for publication Not applicable. Conflict of interest There is no conflict of interest. Availability of data and material Below is the link to the electronic supplementary material. https://doi.org/10.5281/zenodo.7262237 Competing interests All authors declare that they have no conflict of interest, and that the research was conducted according to the compliance with ethical standards. Funding . Not applicable. Authors' contributions Cesar Gálvez, Salvador Flores, Francisco Ramírez y Ramírez, Sergio Campos, Raymundo Rosas-Quijano, Pablo Montoya: Conceptualization, Preparation, Validation, Methodology and Supervision. Cesar Galvez, Sergio Campos: executed the experiments. Cesar Galvez, Salvador Flores, Pablo Montoya: Writing-Reviewing and Editing. All authors read and approved the manuscript. Acknowledgments We thank Dr. Miguel Salvador-Figueroa for all the suggestions and comments on the experimental design. To the Moscamed SENASICA-SADER Program for supplying the biological material. 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M., Akutse, K. S., & Ekesi, S. 2020. Metarhizium anisopliae and Beauveria bassiana : pathogenicity, horizontal transmission, and their effects on reproductive potential of Thaumatotibia leucotreta (Lepidoptera: Tortricidae). Journal of Economic Entomology , 113 (2), 660-668. https://doi.org/10.1093/jee/toz342. Molnár, I., Gibson, D. M., & Krasnoff, S. B. (2010). Secondary metabolites from entomopathogenic Hypocrealean fungi. Natural Product Reports , 27 (9), 1241-1275. https://doi.org/10.1039/C001459C. Montoya, P., Flores, S., Campos, S., Liedo, P., & Toledo, J. (2020). Simultaneous use of SIT plus disseminator devices of Beauveria bassiana enhances horizontal transmission in Anastrepha ludens . Journal of Applied Entomology , 144 (6), 509-518. https://doi.org/10.1111/jen.12766. Novelo-Rincón, L. F., Montoya, P., Hernández-Ortíz, V., Liedo, P., & Toledo, J. (2009). Mating performance of sterile Mexican fruit fly Anastrepha ludens (Dipt., Tephritidae) males used as vectors of Beauveria bassiana (Bals.) Vuill. Journal of Applied Entomology , 133 (9-10), 702-710. https://doi.org/10.1111/j.1439-0418.2009.01427.x. Papadopoulos, N. T. (2008). Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). In J. L. Capinera (Ed.), Encyclopedia of Entomology (Vol. 3, pp. 2318-2322). Springer. Patt, J. M., Chow, A., Meikle, W. G., Gracia, C., Jackson, M. A., Flores, D., Sétamou, M., Dunlap, C. A., Avery, P. B., Hunter, W. B., & Adamczyk, J. J. (2015). Efficacy of an autodisseminator of an entomopathogenic fungus, Isaria fumosorosea , to suppress Asian citrus psyllid, Diaphorina citri , under greenhouse conditions. Biological Control , 88 , 37–45. https://doi.org/10.1016/j.biocontrol.2015.04.014. Peng, G., Jin, K., Liu, Y., & Xia, Y. (2015). Enhancing the utilization of host trehalose by fungal trehalase improves the virulence of fungal insecticide. Applied Microbiology and Biotechnology , 99 (20), 8611-8618. https://doi.org/10.1007/s00253-015-6767-y. Quesada-Moraga, E., Martin-Carballo, I., Garrido-Jurado, I., & Santiago-Álvarez, C. (2008). Horizontal transmission of Metarhizium anisopliae among laboratory populations of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Biological Control , 47 (1), 115-124. https://doi.org/10.1016/j.biocontrol.2008.07.002. Ramírez y Ramírez, F., Salvador-Figueroa, M., Rosas-Quijano, R., Cruz-López, L., Toledo, J., & Quintero-Fong, L. (2022). Sexual performance and survival of males of Ceratitis capitata VIENNA 8 (Diptera: Tephritidae) inoculated with a commercial formulation of Beauveria bassiana. Journal of Applied Entomology , 146 (1-2), 67-76. https://doi.org/10.1111/jen.12935. SENASICA-SAGRAPA (2017). Manual Técnico para las Operaciones Campo de la Campaña Nacional contra Moscas de la Fruta: Sección IV control biológico. Retrieved March 10, 2022, from https://www.gob.mx/cms/uploads/attachment/file/270335/MT_Operaciones_de_campo_CNMF_Secci_n_IV__CB.pdf. Shah, P., & Pell, J. (2003). Entomopathogenic fungi as biological control agents. Applied Microbiology and Biotechnology , 61 (5-6), 413-423. https://doi.org/10.1007/s00253-003-1240-8. Shelly, T. E. (2018). Sexual selection on leks: a fruit fly primer. Journal of Insect Science , 18 (3), 9. https://doi.org/10.1093/jisesa/iey048. Shoukat, R. F., Zafar, J., Shakeel, M., Zhang, Y., Freed, S., Xu, X., & Jin, F. (2020). Assessment of lethal, sublethal, and transgenerational effects of Beauveria bassiana on the demography of Aedes albopictus (Culicidae: Diptera). Insects , 11 (3), 178. https://doi.org/10.3390/insects11030178. Sookar, P., Bhagwant, S., Allymamod, M. N. (2014). Effect of Metarhizium anisopliae on the fertility and fecundity of two species of fruit flies and horizontal transmission of mycotic infection. Journal of Insect Science , 14 (1), 100. http://www.insectscience.org/14.100. Steinkraus, D. C. (2006). Factors affecting transmission of fungal pathogens of aphids. Journal of Invertebrate Pathology , 92 (3),125-131. https://doi.org/10.1016/j.jip.2006.03.009. Stewart, T. P., & Johanson, D. S. (1999). The SPS agreement of the World Trade Organization and plant pest infestation: a case study of the 1997 Mediterranean fruit fly outbreak in Florida. American University International Law Review , 14 (4), 1107-1127. Thaochan, N., & Ngampongsai, A. (2018). Effect of Metarhizium guizhouense infection on mating competition and mate choice of Bactrocera latifrons (Diptera: Tephritidae) . Phytoparasitica 48 , 459-469 . https://doi.org/10.1007/s12600-018-0685-3. Toledo, J., Liedo, P., Flores, S., Campos, S. E., Villaseñor, A., & Montoya, P. (2006). Use of Beauveria bassiana and Metarhizium anisopliae for fruit fly control: A novel approach. In R. L. Sugayama, R. A: Zucchi, S. M. Ovruski & J. Sivinski (Eds.) Proceedings of 7th International Symposium on Fruit Flies of Economic Importance ; Salvador, Brazil. 10-15 September 2006 (pp. 127-132). SBPC. Toledo, J., Campos, S. E., Flores, S., Liedo, P., Barrera, J. F., Villaseñor, A., & Montoya, P. (2007). Horizontal transmission of Beauveria bassiana in Anastrepha ludens (Diptera: Tephritidae) under laboratory and field cage conditions. Journal of Economic Entomology , 100 (2), 291-297. https://doi.org/10.1093/jee/100.2.291. Toledo, J., Flores, S., Campos, S., Villaseñor, A., Enkerlin, W., Liedo, P., Valle, A., & Montoya, P. (2017). Pathogenicity of three formulations of Beauveria bassiana and efficacy of autoinoculation devices and sterile fruit fly males for dissemination of conidia for the control of Ceratitis capitata. Entomologia Experimenatlis et Applicata , 164 (3), 340-349. https://doi.org/10.1111/eea.12608. Usman, M.,Wakil, W., Piñero, J. C.,Wu, S., Toews, M. D., & Shapiro-Ilan, D. I. (2021). Evaluation of locally isolated entomopathogenic fungi against multiple life stages of Bactrocera zonata and Bactrocera dorsalis (Diptera: Tephritidae): laboratory and field study. Microorganisms , 9 , 1791. https://doi.org/10.3390/microorganisms9081791. Vera, M. T., Rodriguez, R., Segura, D. F., Cladera, J. L., & Sutherst, R. W. (2002). Potential geographical distribution of the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), with emphasis on Argentina and Australia. Environmental Entomology , 31 (6), 1009-1022. https://doi.org/10.1603/0046-225X-31.6.1009. Vickers, R. A., Furlong, M. J., White, A., & Pell, J. K. (2004). Initiation of fungal epizootics in diamondback moth populations within a large field cage: proof of concept for auto-dissemination. Entomologia Experimentalis et Applicata , 111 (1), 7–17. https://doi:10.1111/j.0013-8703.2004.00140.x Xia, Y., Clarkson, J. M., & Charnley, A. K. (2002). Trehalose-hydrolysing enzymes of Metarhizium anisopliae and their role in pathogenesis of the tobacco hornworm, Manduca sexta . Journal of Invertebrate Pathology , 80 (3), 139-147. https://doi.org/10.1016/S0022-2011(02)00105-2. Yousef, M., Aranda-Valera, E., & Quesada-Moraga, E. (2018). Lure-and-infect and lure-and-kill devices based on Metarhizium brunneum for spotted wing Drosophila control. Journal of Pest Science , 91 (1), 227-235. https://doi.org/10.1007/s10340-017-0874-8. Zimmermann, G. (2007). Review on safety of the entomopathogenic fungi Beauveria bassiana and Beauveria brongniartii. Biocontrol Science and Technology , 17 (6), 553-596. https://doi.org/10.1080/09583150701309006. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2180398","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":148269893,"identity":"02d3c924-f1cd-4be6-bbaa-6675dd468978","order_by":0,"name":"Cesar Galvez","email":"","orcid":"","institution":"Centro Nacional de Referencia de Control Biológico, SENASICA-DGSV","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Cesar","middleName":"","lastName":"Galvez","suffix":""},{"id":148269894,"identity":"93e483de-64d0-4395-919a-8e5194052f04","order_by":1,"name":"Salvador Flores","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYDAC5jNgirGBvQfM4OEjqIUtB6qFB6j3AFALG/FaJHLAWhgIapFv4z324UONjWz/zLcHH3/MsZNhY2B++OgGHi0Gx/iSZ844lmY843ZessHBbclAh7EZG+fg0yLfY8zM23A4seF2jpnEwW3MQC08bNL4tMi38Rgz/234nzj/5hmQlnrCWhiOAbUwNhxI3HCDB6TlMGEtIL8w9hxLNt54JsfY4Oy24zxszAT8Agyxwww/auxk5x0/Y/igclu1PT9788PHeB2GCZhJUz4KRsEoGAWjAAsAAPcFR7rdUnJnAAAAAElFTkSuQmCC","orcid":"","institution":"Programa Nacional de Moscas de la Fruta SADER-SENASICA","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Salvador","middleName":"","lastName":"Flores","suffix":""},{"id":148269895,"identity":"5a96ffdd-032d-427a-9572-898d85df9880","order_by":2,"name":"Sergio Campos","email":"","orcid":"","institution":"Programa Nacional de Moscas de la Fruta SADER-SENASICA","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sergio","middleName":"","lastName":"Campos","suffix":""},{"id":148269896,"identity":"ce161673-a9e5-4c6a-b620-bd9724d4b96c","order_by":3,"name":"Francisco Ramirez y Ramirez","email":"","orcid":"","institution":"SADER-SENASICA, CDMX","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Francisco","middleName":"Ramirez y","lastName":"Ramirez","suffix":""},{"id":148269897,"identity":"e9fb66ff-1632-46c1-9ccf-5380b554e5ce","order_by":4,"name":"Raymundo Rosas-Quijano","email":"","orcid":"","institution":"Universidad Autónoma de Chiapas","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Raymundo","middleName":"","lastName":"Rosas-Quijano","suffix":""},{"id":148269898,"identity":"0b423586-7c86-475d-a11c-fccea1c3a32a","order_by":5,"name":"Pablo Montoya","email":"","orcid":"","institution":"Programa Nacional de Moscas de la Fruta SADER-SENASICA","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Pablo","middleName":"","lastName":"Montoya","suffix":""}],"badges":[],"createdAt":"2022-10-18 20:14:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2180398/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2180398/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12600-023-01057-y","type":"published","date":"2023-02-21T18:59:27+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":28567189,"identity":"c60b33a0-7222-4496-9636-42a0cf835ee6","added_by":"auto","created_at":"2022-11-02 14:56:17","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":184756,"visible":true,"origin":"","legend":"\u003cp\u003eSurvival (lx) and fecundity (mx) of \u003cem\u003eCeratitis capitata\u003c/em\u003e females after ten days of exposure to \u003cem\u003eBeauveria bassiana\u003c/em\u003e conidia using the auto-dissemination device or infected males by topical method. (a) Females and males inoculated by device; (b) females and males inoculated by topical method; (c) no-inoculated females and males inoculated by device; (d) no-inoculated females and males inoculated by topical method; (e) no inoculated males and females inoculated by device; (f) no inoculated males and females inoculated by topical method; (g) no-inoculated females and males (control).\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-2180398/v1/d3cf903fc197b964f60c0853.png"},{"id":28567190,"identity":"28b2e4fd-1534-4a53-84e2-054f2399a94d","added_by":"auto","created_at":"2022-11-02 14:56:17","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":155667,"visible":true,"origin":"","legend":"\u003cp\u003eSurvival (lx) and fecundity (mx) of \u003cem\u003eCeratitis capitata\u003c/em\u003e females after ten days from exposure with inoculated or no-inoculated males with \u003cem\u003eBeauveria bassiana\u003c/em\u003e conidia. (a) No inoculated sterile male; (b) Inoculated sterile male; (c) No-inoculated fertile males; (d) Inoculated fertile males; (e) no-inoculated sterile and fertile males; (f) inoculated sterile and no inoculated fertile males.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-2180398/v1/8fb3ec4934c3387cc55b6e0a.png"},{"id":44720231,"identity":"5516d363-b517-49c6-8558-462d037f4855","added_by":"auto","created_at":"2023-10-16 19:09:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":624457,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2180398/v1/2f26db08-6cc2-4c2d-9db1-7e41aa35e1b7.pdf"},{"id":28568250,"identity":"14b8c67e-0163-4fab-ae58-01fe7b1bf3a1","added_by":"auto","created_at":"2022-11-02 15:04:17","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1269032,"visible":true,"origin":"","legend":"","description":"","filename":"Datasupplementary.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-2180398/v1/dc3e5e90ef0d6eadc00f18e3.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Horizontal transmission of Beauveria bassiana spores using infected males and inoculation device: impact on survival and fecundity of Ceratitis capitata (Diptera: Tephritidae)","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn natural ecosystems, entomopathogenic fungi (EPF) play an important role in the regulation of insect populations (Goettel et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) and therefore have great potential as biocontrol agents (Dimbi et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Infection by EPF begins with the adherence of the conidium and penetration through the cuticle, followed by the consumption of nutrients present in the insect\u0026rsquo;s hemolymph by the fungus. During mycelial growth, toxins are produced as secondary metabolites, which may cause physiological and behavioral alterations (Xia et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Khachatourians \u0026amp; Qazi, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Moln\u0026aacute;r et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Anderson et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Jin et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The fecundity of infected hosts can be strongly affected during the invasion process, and thus may represent an additional component in the suppression of pest populations (Shoukat et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Mkiga et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Baverstock et al. (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) mention that fungal spore transmission may occur by horizontal transmission among individuals of the same species or by assisted auto-dissemination (using artificial autoinoculation devices). Both modes of transmission have been able to cause fungal infections in some target fruit fly pests (e.g., Quesada-Moraga et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Dimbi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Sookar et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Patt et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Toledo et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Yousef et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In both cases, the horizontal transmission rate has been the key factor determining the spread of the pathogen within host populations (Steinkraus, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Baverstock et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eBeauveria bassiana\u003c/em\u003e (Balsamo) Vuillemin is a facultative fungus with a wide host range found in both temperate and tropical zones (Shah \u0026amp; Pell, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Zimmermann, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The pathogenicity of \u003cem\u003eB. bassiana\u003c/em\u003e has been evaluated in the fruit fly genus \u003cem\u003eAnastrepha\u003c/em\u003e spp. (De la Rosa et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Toledo et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) and in \u003cem\u003eCeratitis capitata\u003c/em\u003e Wiedemann (Diptera: Tephritidae) (Ekesi et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Dimbi et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Toledo et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Flores et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Toledo et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Dias et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2022a\u003c/span\u003e) note that \u003cem\u003eB. bassiana\u003c/em\u003e is one of the most promising and versatile fungus species that can function as a biocontrol agent. Toledo et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and Montoya et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) evaluated this species with the concurrent use of autoinoculation devices and the Sterile Insect Technique (SIT), observing an increase in infected wild fruit flies, and thus proposed an additive or synergistic effect of both technologies.\u003c/p\u003e \u003cp\u003eThe Mediterranean fruit fly, \u003cem\u003eC. capitata\u003c/em\u003e, is a highly polyphagous, multivoltine species that infects more than 353 host fruit species around the world (Papadopoulos, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Currently, this pest has an extensive distribution in tropical and temperate zones, limiting the growth and development of international fresh fruit trade (Stewart \u0026amp; Johanson, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Vera et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Malacrida et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Diamantidis et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Dias et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022b\u003c/span\u003e). In Mexico, the SIT is applied to eradicate \u003cem\u003eC. capitata\u003c/em\u003e transient entries using an integrated pest management (IPM) framework. The sterile insects are provided by the mass-rearing facility located in Metapa, Chiapas, where the insects have been inoculated with \u003cem\u003eB. bassiana\u003c/em\u003e conidia to infect wild flies in the border with Guatemala. The use of autoinoculation devices to disseminate EPF conidia has been another strategy employed to control pest populations (SENASICA-SAGARPA, 2017; Flores et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Toledo et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The selected EPF strains must allow enough time for interactions to occur and conidia to disperse among conspecifics before the insect\u0026rsquo;s death. However, how the infection process affects the sexual competitiveness of males, as well as the resulting fitness of the female conidia recipients, is still not well known. Thus, the objectives of the present work were to determine the effect of the horizontal transmission of \u003cem\u003eB. bassiana\u003c/em\u003e on the survival, fecundity, and fertility of \u003cem\u003eC. capitata\u003c/em\u003e individuals using inoculated adults and auto-inoculation devices. We also determined the impact of the fungus infection on sterile male performance as vectors of EPF. The above will contribute to a better understanding of pathogen-host interactions, and thus reinforce these ecologically oriented control strategies within IPM.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e \u003cem\u003eBiological material.\u003c/em\u003e Males (sterile, fertile) and fertile females of \u003cem\u003eC. capitata\u003c/em\u003e were provided by the Moscamed facility (SADER-SENASICA) located in Metapa, Chiapas, Mexico. After emergence, each sex was kept separately in Plexiglas cages (30 \u0026times; 30 \u0026times; 30 cm) with water and food (a 3: 1 mixture of sugar: hydrolyzed protein [MP Biomedicals, LLC, Santa Ana, California, USA]) until they reached sexual maturity. Laboratory conditions were maintained at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, 60\u0026thinsp;\u0026plusmn;\u0026thinsp;10% R.H, and a photoperiod of 12:12 h light: dark. All tests were carried out inside a cage 3 m in diameter \u0026times; 2 m high made of saran screen (20 by 20 mesh) in an isolated room inside the Moscamed facility to reinforce biological safety measures.\u003c/p\u003e \u003cp\u003eThe \u003cem\u003eB. bassiana\u003c/em\u003e strain was formulated with 2 \u0026times; 10\u003csup\u003e9\u003c/sup\u003e conidia per g with Celite 400 as inert material (Laboratorio de Organismos Ben\u0026eacute;ficos, Hongos, Insectos y Nematodos, Talisman, Chiapas, Mexico). The viability of the fungus was determined before each test according to the microculture technique (Jim\u0026eacute;nez, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1992\u003c/span\u003e) and with observations of germinated conidia at 40\u0026times; using an optical microscope (Dialux 20 EB; Leitz, Wetzlar, Germany). The conidia were considered viable when the length of the germ tube was greater than the diameter. The evaluations were carried out with batches of conidia with a recorded viability of \u0026ge;\u0026thinsp;90%.\u003c/p\u003e \u003cp\u003e \u003cem\u003eInoculation methods\u003c/em\u003e. Adults of \u003cem\u003eC. capitata\u003c/em\u003e were infected with \u003cem\u003eB. bassiana\u003c/em\u003e conidia either topically or using the inoculation device method: \u003cem\u003e1) Topical inoculation.\u003c/em\u003e We applied 0.05 g of the \u003cem\u003eB. bassiana\u003c/em\u003e formulation to 50 fertile or sterile male or female \u003cem\u003eC. capitata\u003c/em\u003e in a 50 ml Falcon conical centrifuge tube (29 mm diameter \u0026sdot; 115 mm height), which was shaken gently for 10s for a homogeneous inoculation of all individuals. \u003cem\u003e2) Inoculation with PVC device.\u003c/em\u003e The device was constructed using a PVC tube (9 cm in diameter \u0026sdot; 10 cm high) with two perforations of 1 cm in diameter in the lower part, and 100 \u0026sdot; 10 mm Petri dish bottoms were used as lids on both ends. The interior of the tube was lined with yellow plush fabric (felt), similarly to Toledo et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), which was homogeneously impregnated with 2 g of the conidia formulation. Groups of 50 females or males were placed inside test tubes (25 \u0026sdot; 150 mm) and then introduced through the perforations on the lower part of the PVC device. The insects were recovered after 10 min, which was enough time to ensure contact with the fungal conidia. Inoculated adults were separated by sex and kept in Plexiglas cages (30 \u0026sdot; 30 \u0026sdot; 30 cm) for 20 min to allow them to remove excess conidia by grooming.\u003c/p\u003e \u003cp\u003eThe effect of \u003cem\u003eB. bassiana\u003c/em\u003e conidia on the fecundity, fertility, and survival of \u003cem\u003eC. capitata\u003c/em\u003e adults was determined through the following tests:\u003c/p\u003e \u003cp\u003ea) \u003cem\u003eEffect of inoculation method.\u003c/em\u003e The effect of \u003cem\u003eB. bassiana\u003c/em\u003e on \u003cem\u003eC. capitata\u003c/em\u003e was determined by direct inoculation of females or by horizontal transmission through inoculated males. The combinations evaluated were: inoculated males\u0026thinsp;+\u0026thinsp;inoculated females, inoculated males\u0026thinsp;+\u0026thinsp;non-inoculated females, non-inoculated males\u0026thinsp;+\u0026thinsp;inoculated females. Fungal conidia were applied to adults either by topical inoculation or with the PVC device. A treatment with non-inoculated males\u0026thinsp;+\u0026thinsp;non-inoculated females was included as a control treatment. This resulted in seven treatments, three combinations \u0026times; two inoculation methods\u0026thinsp;+\u0026thinsp;control.\u003c/p\u003e \u003cp\u003eFor each treatment, 10 fertile fly pairs were placed in plastic containers of 1.75 L (17.3 cm high, 21.2 \u0026times; 11.3 cm) provided with water and food (3:1 mixture of sugar: hydrolyzed protein). The container was placed in a horizontal position and kept under laboratory conditions for 10 days. As oviposition substrate, the opening of the plastic container was covered with a piece of white lycra fabric (Dorian Gray Likra Pantyhose). Laid eggs fell into a petri dish with tap water placed below the container opening. Each container was one experimental unit, and each repetition was carried out with a different production batch of \u003cem\u003eC. capitata.\u003c/em\u003e We performed five replicates per treatment for horizontal transmission.\u003c/p\u003e \u003cp\u003e \u003cem\u003eb) Inoculation of sterile males.\u003c/em\u003e The effect of infected sterile males on the survival, fecundity, and fertility of fertile females was compared with that of inoculated fertile males and the interaction between fertile and inoculated sterile males. The treatments were (1) inoculated sterile males and (2) non-inoculated sterile males, both interacting with fertile males. The topical method was used to apply the conidia to both sterile and fertile males.\u003c/p\u003e \u003cp\u003eAccording to each treatment, 10 males and 10 females were placed in plastic containers of 1.75 L as described above. In the fertile-sterile male treatments, 5 sterile males (inoculated or healthy) and 5 fertile males were placed with 10 fertile females. Each container was an experimental unit with six replicates per treatment, each one with a different production batch of \u003cem\u003eC. capitata.\u003c/em\u003e Fecundity, fertility, mortality, and mycosis were estimated as follows:\u003c/p\u003e \u003cp\u003e \u003cem\u003eFecundity and egg fertility.\u003c/em\u003e Eggs were collected in a Petri dish bottom (150 \u0026sdot; 20 mm) with 200 ml of sterile water placed under the oviposition substrate. For each treatment, eggs were recovered daily using a piece of black organza cloth (10 \u0026sdot; 10 cm) and were aligned and counted using a stereomicroscope (Nikon, SMZ645, USA). A sample of 100 eggs was placed in an incubation chamber (plastic Petri dish of 100 \u0026sdot; 10 mm, where a piece of black cloth was placed on a piece of 2-mm-thick moistened filter paper). After four days, the hatched eggs were counted using a stereomicroscope with a 4\u0026sdot; objective. Egg fertility was expressed as percentage of hatched eggs.\u003c/p\u003e \u003cp\u003e \u003cem\u003eMortality and fungal sporulation.\u003c/em\u003e The number of dead adults in each treatment was recorded daily. Dead adults were disinfected for 20 sec in a 1% sodium hypochlorite solution and then washed three times with sterile distilled water to eliminate saprophytic microorganisms. Subsequently, the specimens were placed in a humid chamber (sterile wet filter paper in 100 \u0026times; 10 mm plastic Petri dishes) sealed with Parafilm (Bemis Company, Inc. Neenah, WI, 54956) and kept under laboratory conditions. After five days, the flies were examined under a stereomicroscope to verify the presence of mycosis.\u003c/p\u003e \u003cp\u003e \u003cem\u003eStatistical analysis\u003c/em\u003e. In each test, survival curves over the course of the ten days of evaluation were compared using a log-rank test. Fecundity, number of eggs per live female per day, was log-transformed to obtain homoscedasticity prior to performing an analysis of variance (ANOVA), and mean comparisons were performed with a Tukey test (α\u0026thinsp;=\u0026thinsp;0.05). Fertility, expressed as the proportion of hatched eggs, was analyzed with a GLM with binomial distribution and a logit link function. Treatment means were compared by contrasts with a significance level of 95%.\u003c/p\u003e \u003cp\u003eIn the case of horizontal transmission of \u003cem\u003eB. bassiana\u003c/em\u003e calculated in treatments with inoculated and non-inoculated adults, mycosis in recipient flies was analyzed using a generalized linear model (GLM) with binomial distribution and a logit link function, and treatments were compared by orthogonal contrasts (α\u0026thinsp;=\u0026thinsp;0.05). All analyses were conducted in JMP\u0026reg; 11.0.0 (JMP\u0026reg; SAS Institute, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.jmp.com\u003c/span\u003e\u003cspan address=\"http://www.jmp.com\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003ea) \u003cem\u003eInoculation method\u003c/em\u003e\u003c/p\u003e \u003cp\u003eSurvival of females (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;217.98, d.f. = 6, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and males (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;189.34, d.f. = 6, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) differed significantly between treatments. The survival of directly inoculated females was reduced to at least 20% at 5 days. In treatments with inoculated males, the survival of non-inoculated females showed a gradual reduction throughout the 10 days. In the control treatment, the mortality was lower than 20% after 10 days. The survival curve of females did not differ between topical inoculation and the use of an inoculation device (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe number of eggs per female was higher at day 3 and 4 in each treatment, with the control treatment having the highest number of eggs (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Fecundity was significantly affected by treatment (F\u0026thinsp;=\u0026thinsp;18.58, d.f. = 6, 24, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The control treatment had the highest fecundity, and the treatments with directly inoculated females showed the lowest fecundity. When comparing the inoculation methods, the treatments with topical inoculation showed lower fecundity than those with the device method (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFecundity and egg fertility of \u003cem\u003eCeratitis capitata\u003c/em\u003e females in treatments using inoculated insects with \u003cem\u003eBeauveria bassiana\u003c/em\u003e conidia, applied by topical or device method.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreated Sex\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMethod of inoculation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eFecundity\u003c/p\u003e \u003cp\u003e(Eggs/♀ \u0026plusmn; S. E.)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eEgg Fertility\u003c/p\u003e \u003cp\u003e(% Eclosion\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E.)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eTransmission\u003c/p\u003e \u003cp\u003e(% mycosis\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E.)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e♂-♀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDevice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.3\u0026thinsp;\u0026plusmn;\u0026thinsp;12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ec\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTopical\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50.1\u0026thinsp;\u0026plusmn;\u0026thinsp;10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e68.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e♂\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDevice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e155.8\u0026thinsp;\u0026plusmn;\u0026thinsp;34.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.0\u0026thinsp;\u0026plusmn;\u0026thinsp;12.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTopical\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e148.8\u0026thinsp;\u0026plusmn;\u0026thinsp;34.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ebc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e70.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ec\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e74.5\u0026thinsp;\u0026plusmn;\u0026thinsp;11.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e♀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDevice\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72.6 \u0026plusmn; 9.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ecd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e78.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e76.0\u0026thinsp;\u0026plusmn;\u0026thinsp;11.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTopical\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e50.0 \u0026plusmn; 9.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e77.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80.0\u0026thinsp;\u0026plusmn;\u0026thinsp;11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e243.1\u0026thinsp;\u0026plusmn;\u0026thinsp;29.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e86.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eMeans followed by the same letter in each column are not significantly different (α\u0026thinsp;=\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe fertility of \u003cem\u003eC. capitata\u003c/em\u003e was significantly affected in treatments with the fungus spore (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;399.59, d.f. = 6, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001), with the control having the highest proportion of hatched eggs. The treatments with directly inoculated males showed the lowest egg hatching percentage. The treatments with topical inoculation of males showed the lowest fertility values (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFungal sporulation in inoculated adults varied from 94 to 98% between males and females in the different treatments (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;8.23, d.f. = 7, P\u0026thinsp;=\u0026thinsp;0.312). The horizontal transmission of \u003cem\u003eB. bassiana\u003c/em\u003e did not differ significantly between treatments where inoculated and non-inoculated individuals coexisted (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3.76, d.f. = 3, P\u0026thinsp;=\u0026thinsp;0.288) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eb) Infection of sterile males\u003c/h2\u003e \u003cp\u003eFemale survival (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;217.59, df\u0026thinsp;=\u0026thinsp;5, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) was significantly different between treatments. The survival curve of females in the treatments with inoculated males was significantly reduced compared to treatments with non-inoculated males (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;216.88, d.f. = 1, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The survival curve of females exposed to inoculated males (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.47, d.f. = 2, P\u0026thinsp;=\u0026thinsp;0.789) and non-inoculated males (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;1.15, d.f. = 2, P\u0026thinsp;=\u0026thinsp;0.64) was not affected by the male strain used (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe number of eggs per female per day showed the highest peak between days 3 and 4 in each treatment. In the treatments with non-inoculated males, the number of eggs per female exhibited a gradual decrease, while there was greater variation after day 6 in the treatments with inoculated males (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The total fecundity for the entire period was significantly lower in treatments with inoculated males than in treatments with non-inoculated males (F\u0026thinsp;=\u0026thinsp;17.25, d.f. = 1.25, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Fecundity did not differ significantly between the treatments with sterile, fertile, or sterile and fertile males (F\u0026thinsp;=\u0026thinsp;0.82, d.f. = 2, 25, P\u0026thinsp;=\u0026thinsp;0.450) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFecundity and egg fertility of \u003cem\u003eCeratitis capitata\u003c/em\u003e females in field cage tests, using \u003cem\u003eBeauveria bassiana\u003c/em\u003e inoculated sterile and no-inoculated fertile males.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale strain\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eFecundity\u003c/p\u003e \u003cp\u003e(Eggs/♀ \u0026plusmn; S.E.)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eEgg Fertility\u003c/p\u003e \u003cp\u003e(% Eclosion\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E.)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eTransmission\u003c/p\u003e \u003cp\u003e(% mycosis\u0026thinsp;\u0026plusmn;\u0026thinsp;S.E.)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFertile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo-inoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e226.0\u0026thinsp;\u0026plusmn;\u0026thinsp;24.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eInoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e148.1\u0026thinsp;\u0026plusmn;\u0026thinsp;40.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e72.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ec\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e73.3\u0026thinsp;\u0026plusmn;\u0026thinsp;11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSterile-Fertile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo inoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e232.9\u0026thinsp;\u0026plusmn;\u0026thinsp;15.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e64.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ed\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eInoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e150.6\u0026thinsp;\u0026plusmn;\u0026thinsp;30.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e81.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e73.3\u0026thinsp;\u0026plusmn;\u0026thinsp;11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSterile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo inoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e211.0\u0026thinsp;\u0026plusmn;\u0026thinsp;29.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eInoculated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e125.5\u0026thinsp;\u0026plusmn;\u0026thinsp;39.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e73.3\u0026thinsp;\u0026plusmn;\u0026thinsp;11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003ea\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eMeans followed by the same letter in each column are not significantly different (α\u0026thinsp;=\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe egg fertility of \u003cem\u003eC. capitata\u003c/em\u003e was significantly affected by treatment (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;724.7, d.f. = 3, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Inoculation of fertile males significantly reduced the percentage of hatched eggs. When inoculated or non-inoculated sterile males interacted with fertile males and females, egg fertility was lower than with non-inoculated sterile males (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHorizontal transmission to females did not differ significantly between the three treatments that included males inoculated with \u003cem\u003eB. bassiana\u003c/em\u003e, with a mycosis percentage of 73.3% in each treatment (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eAs expected, the survival and fecundity of \u003cem\u003eC. capitata\u003c/em\u003e were affected by the infection of \u003cem\u003eB. bassiana\u003c/em\u003e. The use of inoculated sterile males or autoinoculation devices resulted in the horizontal transmission of \u003cem\u003eB. bassiana\u003c/em\u003e to untreated insects. Both inoculation methods allowed the successful interaction between treated and untreated insects, allowing thus a horizontal transmission between them, which provides a better understanding of how infections by fungi, such as \u003cem\u003eB. bassiana\u003c/em\u003e, affect the survival and reproduction of contaminated insects. This undoubtedly will lead to the implementation of more effective and timelier IPM strategies (Shoukat et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) for the control of fruit fly populations. Our results support the proposal to simultaneously use the SIT and EPF as in Montoya et al. (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Furthermore, devices for disseminating EPF conidia are inexpensive, easy to apply, and effective (Toledo et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Yousef et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur results showed a range of 65\u0026ndash;80% horizontal transmission of \u003cem\u003eB. bassiana\u003c/em\u003e conidia to untreated \u003cem\u003eC. capitata\u003c/em\u003e adults. These results agree with Chergui et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), who recorded a maximum of 51.67 and 73.33% mortality on the 15th day of a contact bioassay in males and females, respectively. The success of EPF as a control agent depends mainly on the possibility of transmitting the infection among conspecifics, as well as on the behavioral response of the insect species during the host-pathogen interaction (Baverstock et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Dimbi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). In the case of fruit flies, fungal transmission is enhanced by the mating behavior displayed by the adults. Fruit fly males form \u003cem\u003eleks\u003c/em\u003e (i.e., groups of males calling for matings [Shelly, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Thaochan \u0026amp; Ngampongsai, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e]) and compete with each other to copulate with the females present. This behavior generates several interactions that increase the chances of successful fungal transmission among conspecifics. We observed that the horizontal transmission of conidia was higher with the topical method than with the auto-inoculation device, which could be due to the topical method resulting in a larger number of conidia per fly than the device method. Chergui et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) indicate that different methods of \u003cem\u003eB. bassiana\u003c/em\u003e application produce different results. According to Kaneshiro et al. (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), a larger amount of conidia could generate female rejection of sterile males, although the excess of EPF could be reduced by grooming (Baverstock et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Moreover, grooming allows conidia to be deposited in preferred gathering sites for adults, for example, during \u003cem\u003elek\u003c/em\u003e formation (Arita \u0026amp; Kaneshiro, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Thaochan \u0026amp; Ngampongsai, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe survival of treated adults was reduced from 7 to 10 days. During this period, females can oviposit before dying; however, their reproductive potential was highly reduced by the EPF infection (see Toledo et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). The effect of \u003cem\u003eB. bassiana\u003c/em\u003e of reducing the fecundity, fertility, and survival of \u003cem\u003eC. capitata\u003c/em\u003e females in this study was due to the physiological alterations derived from the pathogenic infection (Jin et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Usman et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). For example, the depletion of sugar and other compounds in the insect hemolymph by EPF (e.g., Xia et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Jin et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Peng et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) has serious consequences on the fitness parameters of the host insects (Jin et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The observed reduction in the oviposition rate and fertility of infected females has been highly associated with the action of EPF, which negatively impacts insect populations as a resulting compensation for their delayed mortality (Dimbi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The adverse effects on fecundity can also be caused by the antifeedant activity of \u003cem\u003eB. bassiana\u003c/em\u003e during the invasive process (Ekesi, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Our results showed that fecundity was lower in directly infected females than in females infected by inoculated fertile males. We also observed a negative effect on egg hatching when fertile males were inoculated than when inoculated sterile males interacted with non-inoculated fertile males.\u003c/p\u003e \u003cp\u003eThe use of insects as vectors of biocontrol agents has been proposed as a strategy to improve biological control programs through the horizontal transmission of pathogens (Vickers et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Llacer et al., 2013; Diouf et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The application of this strategy in fruit fly programs allows the release of inoculated sterile males to disseminate conidia into wild populations of \u003cem\u003eC. capitata\u003c/em\u003e (Toledo et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Recently, Diouf et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) introduced the term \u0026ldquo;boosted SIT\u0026rdquo; to refer to the use of sterile insects as vectors of biocides to trigger an epizootic in wild populations. For successful results, quality parameters, such as dispersion and competitiveness, of sterile males must remain unaffected after inoculation (Novelo-Rincon et al., 2009; Ram\u0026iacute;rez y Ram\u0026iacute;rez et al., 2020), at least long enough to interact with a wild population.\u003c/p\u003e \u003cp\u003eIn conclusion, the inoculation of \u003cem\u003eB. bassiana\u003c/em\u003e conidia using a device or the topical method was effective in causing mycosis in adult \u003cem\u003eC. capitata\u003c/em\u003e, and the horizontal transmission of fungal spores significantly reduced the fecundity, egg fertility, and survival of \u003cem\u003eC. capitata\u003c/em\u003e females. These results support the proposal of including this strategy in IPM programs for the suppression of Mediterranean fruit fly populations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe research meets ethical guidelines and adheres to the legal requirements of the study country. This research does not involve human subjects.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere is no conflict of interest.\u003c/p\u003e\n\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBelow is the link to the electronic supplementary material.\u003c/p\u003e\n\u003cp\u003ehttps://doi.org/10.5281/zenodo.7262237\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors declare that they have no conflict of interest, and that the research was conducted according to the compliance with ethical standards.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCesar G\u0026aacute;lvez, Salvador Flores, Francisco Ram\u0026iacute;rez y Ram\u0026iacute;rez, Sergio Campos, Raymundo Rosas-Quijano, Pablo Montoya: Conceptualization, Preparation, Validation, Methodology and Supervision. Cesar Galvez, Sergio Campos: executed the experiments. Cesar Galvez, Salvador Flores, Pablo Montoya: Writing-Reviewing and Editing. All authors read and approved the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Dr. Miguel Salvador-Figueroa for all the suggestions and comments on the experimental design. To the Moscamed SENASICA-SADER Program for supplying the biological material. This research was supported by the National Fruit Fly Program DGSV-SENASICA-SADER, Mexico.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAnderson, R. D., Bell, A. S., Blanford, S., Paaijmans, K. P., \u0026amp; Thomas, M. B. (2011). 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(2018). Effect of \u003cem\u003eMetarhizium guizhouense\u003c/em\u003e infection on mating competition and mate choice of \u003cem\u003eBactrocera latifrons\u003c/em\u003e (Diptera: Tephritidae)\u003cem\u003e. \u003c/em\u003e\u003cem\u003ePhytoparasitica 48\u003c/em\u003e, 459-469\u003cem\u003e.\u003c/em\u003e https://doi.org/10.1007/s12600-018-0685-3.\u003c/li\u003e\n\u003cli\u003eToledo, J., Liedo, P., Flores, S., Campos, S. E., Villase\u0026ntilde;or, A., \u0026amp; Montoya, P. (2006). Use of \u003cem\u003eBeauveria bassiana \u003c/em\u003eand \u003cem\u003eMetarhizium anisopliae \u003c/em\u003efor fruit fly control: A novel approach. In R. L. Sugayama, R. A: Zucchi, S. M. Ovruski \u0026amp; J. Sivinski (Eds.) \u003cem\u003eProceedings of 7th International Symposium on Fruit Flies of Economic Importance\u003c/em\u003e; Salvador, Brazil. 10-15 September 2006 (pp. 127-132). SBPC.\u003c/li\u003e\n\u003cli\u003eToledo, J., Campos, S. E., Flores, S., Liedo, P., Barrera, J. 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Review on safety of the entomopathogenic fungi \u003cem\u003eBeauveria bassiana\u003c/em\u003e and \u003cem\u003eBeauveria brongniartii.\u003c/em\u003e \u003cem\u003eBiocontrol Science and Technology\u003c/em\u003e, \u003cem\u003e17\u003c/em\u003e(6), 553-596. https://doi.org/10.1080/09583150701309006.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"phytoparasitica","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pypa","sideBox":"Learn more about [Phytoparasitica](http://link.springer.com/journal/12597)","snPcode":"12600","submissionUrl":"https://submission.nature.com/new-submission/12600/3","title":"Phytoparasitica","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Beauveria bassiana, Ceratitis capitata, entomopathogenic fungus, sterile insect technique, integrated pest management.","lastPublishedDoi":"10.21203/rs.3.rs-2180398/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2180398/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe mode of transmission of fungus spores (horizontal transmission or assisted auto-dissemination) directly influences the effectiveness of a fungal pathogen when used as a control agent. Fungal infections cause physiological alterations leading to the host's death. During this process, the fungus uses the energy reserves in the hemolymph of insects, affecting the development and performance of individuals and, therefore, the demographic features of their populations. In this work, we evaluated topical inoculation and an auto-disseminator device in the transmission of \u003cem\u003eBeauveria bassiana\u003c/em\u003e (Balsamo) Vuillemin conidia to \u003cem\u003eCeratitis capitata\u003c/em\u003e Wiedemann (Diptera: Tephritidae). Survival and fecundity were negatively affected by the action of the fungus, and mortality was influenced by the inoculation method. Inoculated sterile males were as competitive as untreated males and reduced the fecundity and survival of females. We conclude that the pathogenic action of \u003cem\u003eB. bassiana\u003c/em\u003e reduces the survival and fecundity parameters of \u003cem\u003eC. capitata\u003c/em\u003e infected by horizontal transmission, while the behavioral response of treated sterile males is similar to that of sterile-fertile untreated males. We discuss the potential use of this strategy as part of the pest management of \u003cem\u003eC. capitata\u003c/em\u003e.\u003c/p\u003e","manuscriptTitle":"Horizontal transmission of Beauveria bassiana spores using infected males and inoculation device: impact on survival and fecundity of Ceratitis capitata (Diptera: Tephritidae)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-11-02 14:56:12","doi":"10.21203/rs.3.rs-2180398/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2022-11-22T20:00:33+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2022-11-14T08:37:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"d34aed38-355f-4331-8c98-33e9ad29b804","date":"2022-11-03T06:16:06+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2022-11-02T07:59:04+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2022-11-01T13:48:29+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2022-10-31T12:16:08+00:00","index":"","fulltext":""},{"type":"submitted","content":"Phytoparasitica","date":"2022-10-18T20:00:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"phytoparasitica","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pypa","sideBox":"Learn more about [Phytoparasitica](http://link.springer.com/journal/12597)","snPcode":"12600","submissionUrl":"https://submission.nature.com/new-submission/12600/3","title":"Phytoparasitica","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"5aca05b9-fd0a-4817-aa7e-ea699d2ba73d","owner":[],"postedDate":"November 2nd, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-10-16T19:05:34+00:00","versionOfRecord":{"articleIdentity":"rs-2180398","link":"https://doi.org/10.1007/s12600-023-01057-y","journal":{"identity":"phytoparasitica","isVorOnly":false,"title":"Phytoparasitica"},"publishedOn":"2023-02-21 18:59:27","publishedOnDateReadable":"February 21st, 2023"},"versionCreatedAt":"2022-11-02 14:56:12","video":"","vorDoi":"10.1007/s12600-023-01057-y","vorDoiUrl":"https://doi.org/10.1007/s12600-023-01057-y","workflowStages":[]},"version":"v1","identity":"rs-2180398","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2180398","identity":"rs-2180398","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}