Abstract The objective of this study was to evaluate biological alternatives for the management of coffee berry borer (Hypothenemus hampei Ferrari), on individuals inside brocaded fruits, its effect on the total yield (kg ha-1), economic feasibility of each treatment evaluated. through a partial budget analysis, dominance analysis and marginal rate of return in Jalapa, Nicaragua. A quantitative, applied study was carried out, with a Randomized Complete Block experimental design (BCA), with six treatments and four repetitions, three biological products, one chemical, a combination of chemical-cultural management (trap) and an absolute control, whose variables were subjected to an analysis of variance and separation of means by Tukey test (0.05). It was determined that there was no significant effect of the treatments on the borer control, the pest’s affectation increases the fruit develops, causing losses between 20 and 78 % throughout the cycle. The economic analysis showed that all treatments are profitable, having a high marginal rate of return, The Curador® treatment is the one with the highest net benefit, followed by trap plus Curador® and Bea Blue®, constituting alternatives for the biological management of the borer.

Resumen El presente estudio tuvo como objetivo evaluar alternativas biológicas para el manejo de la broca de café (Hypothenemus hampei Ferrari), sobre la población de individuos dentro de frutos brocados, su efecto en el rendimiento total (kg/ha), factibilidad económica de cada tratamiento evaluado a través de un análisis de presupuesto parcial, análisis de dominancia y tasa de retorno marginal en Jalapa, Nicaragua. Se realizó un estudio de tipo cuantitativo, aplicado, con un diseño experimental de Bloques Completos al Azar (BCA), con seis tratamientos y cuatro repeticiones, tres productos biológicos, un químico, una combinación de químico-manejo cultural (trampa) y un testigo absoluto, cuyas variables fueron sometidas a un análisis de varianza y separación de medias mediante la prueba de Tukey (0,05), se determinó que no hubo efecto significativo de los tratamientos en el control de la broca, las afectaciones de la plaga se incrementan a medida que se desarrolla el fruto provocando pérdidas de entre el 20 y 78 % en todo el ciclo. El análisis económico mostró que todos los tratamientos son rentables, al tener una tasa de retorno marginal alta, el tratamiento Curador® es el de mayor beneficio neto, seguido de trampa más Curador® y Bea Blue®, constituyéndose como alternativas de manejo biológico de la broca.

Coffee berry borer, Hypothenemus hampei (Ferrari, 1867), is a serious insect pest of organic coffee plantation in Säo Tomé Island. To date, limited information regarding the seasonal phenology of this pest species on the islands limits the implementation of integrated pest management (IPM) programmes. As part of a coffee farmer training programme, three attractants were evaluated in red vs. transparent traps to assess olfactory and visual stimuli. The experiment was delineated in a split-block design with three types of attractants: commercial ethanol + 40 g of ripe Robusta coffee (A1), proportion 3:1 methanol and ethanol (A2), and commercial ethanol + 10 g of ground roasted Arabica coffee (A3); and two home-made transparent (D1) and red (D2) traps. The results showed that there was significant interaction between the trap model and the attractant for borer capture. The transparent trap baited with methanol and ethanol exhibited the best result with an average of 14.3 ± 5.4 adults/trap/week. Transparent traps baited captured more borers and largest numbers of beetles were trapped late May through September. In short, home-made traps alone are not effective for controlling the coffee berry borer, but they are useful in monitoring this species.

ABSTRACT Among the management and control tactics of the coffee berry borer Hypothenemus hampei (Ferrari), there is the use of entomopathogenic fungi. Due to the importance of prospecting isolates of entomopathogenic fungi for the control of the coffee berry borer, the objective of this study was to evaluate the efficiency of 26 isolates of entomopathogenic fungi in the control of this insect pest in the laboratory. The coffee berry borers were immersed in a solution adjusted to the concentration of 1 to 3 × 108 conidia/mL of each isolate and the control treatment (sterilized water). After seven days total mortality and confirmed mortality were evaluated. The isolates that caused the highest mortality and two commercial isolates were selected for evaluation of lethal concentration (LC50 and LC90) and lethal time (LT50 and LT90). Coffee berry borers were treated at different conidia concentrations for lethal concentration to assess total and confirmed mortality. For a lethal time, the coffee berry borers were treated at the concentration of 108 conidia/mL of the selected isolates, and, after two days and every 24 hours until the eighth day, the number of dead individuals was verified. Among the 26 isolates evaluated, 24 presented mortality higher than the control treatment, and three presented mortality higher than 85%. In the LC50 and LC90 assays, the IBCB 353 and IBCB 364 isolates were more lethal to H. hampei. In the LT50 and LT90 assays, the IBCB 66 and IBCB 353 isolates caused lethality in a shorter time.

Resumen Las hormigas presentes en los cafetales podrían estar ejerciendo un control natural de la broca del café (BDC), como es el caso de Hypothenemus hampei al depredar a las poblaciones de esta plaga, que quedan en frutos abandonados al final de cada cosecha. El objetivo de esta investigación fue evaluar la capacidad depredadora de dos especies de hormigas (Solenopis picea y Crematogaster crinosa) sobre la broca del café con un atrayente en experimentos de campo. Para cada especie, se establecieron diez unidades experimentales (UE) con diez granos de café infestados con broca. Los granos infestados fueron colocados dentro de una malla de anjeo y colgados en árboles de café con un nido de hormigas. Para atraer las hormigas hacia los granos brocados estos fueron asperjados con una solución atrayente. Como testigo absoluto se usaron los mismos tratamientos con exclusión de hormigas y como testigo relativo granos brocados sin atrayente. La tasa de depredación de S. picea después de 24 h fue de 78,3%, seguida por C. crinosa con 34,3%. El cebo atrayente incrementó en un 22% la atracción de hormigas y depredación de estados biológicos de broca en cada UE, frente al tratamiento no asperjado. Los resultados muestran que las especies de hormigas evaluadas redujeron las poblaciones residuales de H. hampei; S. picea más que C. crinosa.

Abstract The ants present in coffee plantations could be exerting a natural control of the Coffee Berry Borer (CBB), Hypothenemus hampei by preying on the populations of this pest, in fruits abandoned at the end of each harvest. The objective of this research was to evaluate the predatory capacity of two species of ants (Solenopis picea and Crematogaster crinosa) on the CBB with an attractant in field experiments. For each species, ten Experimental Units (EU) containing ten coffee beans infested with CBB were established. The infested grains were placed inside a plastic mesh and hung in trees with that had an ant nest. To attract the ants to the coffee berries, the berries were sprayed with an attractant solution. The same treatments were used with exclusion of ants as absolute control and infested berries without attractant as relative control. The predation rate of S. picea after 24 h was 78.3%, followed by C. crinosa with 34.3%. The attractant bait increased an average of 22% the attraction of ants and predation of biological stages of CBB in each EU, compared to the non-sprayed treatment. The results show that the evaluated ant species reduced the residual populations of H. hampei; S. picea more than C. crinosa.

Resumen El control genético como método de control que puede combinarse con otros métodos, ha sido poco estudiado como alternativa económicamente viable y de fácil adopción en el caso de la broca del café Hypothenemus hampei (Ferrari), plaga que ocasiona las mayores pérdidas económicas al cultivo de café en Colombia. Los esfuerzos por reducir estas pérdidas económicas se han enfocado principalmente en el manejo integrado de la plaga a través de prácticas culturales, químicas y biológicas, que han logrado mantener su infestación por debajo del 5% que representa alrededor del 7% de los costos de producción del café. El empleo de variedades resistentes aumenta la eficiencia del manejo integrado debido a que permite reducir el número de individuos por generación y prolonga su periodo de desarrollo. Esto disminuye el crecimiento de la población y mantiene los insectos por debajo de los niveles de daño económico. Esta revisión presenta información del origen de la broca del café, su dispersión, biología y ciclo de vida, y las pérdidas económicas ocasionadas por la plaga. Se hizo énfasis en los estudios de resistencia de la planta de café contra la broca mediante antibiosis y antixenosis. En el caso de antibiosis, se recopiló la identificación de las fuentes de resistencia del género Coffea, así como la utilización de introducciones etíopes de Coffea arabica y de accesiones de C. liberica de la Colección Colombiana de Café (CCC) como progenitores masculinos para el desarrollo de una variedad con efecto de antibiosis contra la broca. En el caso de antixenosis, se compilaron los estudios pertinentes realizados en especies y variedades de café respecto a H. hampei.

Abstract Genetic control as a control method that can be combined with other methods, has been little studied as an economically viable and easy-to-adopt alternative in the case of the coffee berry borer, Hypothenemus hampei (Ferrari), the pest that causes the greatest economic losses to coffee crop in Colombia. Efforts to reduce these economic losses have been mainly focused on the integrated pest management of the pest through cultural, chemical, and biological practices, which have managed to maintain its infestation below 5%, which represents around 7% of coffee production costs. Genetic control can be successfully combined with the other control methods currently being used. The use of resistant varieties increases the efficiency of integrated pest management because it reduces the number of individuals per generation and prolongs their development period. This reduces population growth and keeps insects below economic injury levels. This review presents information on the origin of the coffee berry borer, its dispersal, biology and life cycle, and the economic losses caused by this pest. Emphasis was placed on studies of coffee plant resistance to CBB through antibiosis and antixenosis. In the case of antibiosis, the identification of sources of resistance in the genus Coffea is compiled, as well as the use of Ethiopian introductions of Coffea arabica and accessions of C. liberica from the Colombian Coffee Collection (CCC) as male progenitors for the development of a coffee variety with antibiosis effect against the CBB. In the case of antixenosis, the relevant studies carried out on coffee species and varieties with respect to H. hampei are compiled.

Resumo O objetivo deste trabalho foi desenvolver um novo método para extração de adultos e larvas de Hypothenemus hampei de frutos de café (Coffea arabica). O extrator é composto por um conjunto de dois recipientes plásticos, um encaixado sobre o outro: o de cima serve para colocar os frutos dentro e o de baixo, como base para a captura de adultos e larvas. A eficiência do extrator foi comparada com a do método de dissecação (controle). O número de adultos vivos, adultos mortos e larvas não difere significativamente entre os dois métodos avaliados. O extrator desenvolvido reduz o trabalho manual, é de custo acessível e é eficaz para a captura de adultos e larvas de H. hampei.

Abstract The objective of this work was to develop a new method for extracting Hypothenemus hampei adults and larvae from coffee (Coffea arabica) fruits. The extractor consists of a set of two plastic containers, with one fit on top of the other: the one on top is used to place the fruits inside and the one on the bottom, as a base to capture adults and larvae. The efficiency of the extractor was compared with that of the dissection method (control). The number of live adults, dead adults, and larvae does not differ significantly between the two evaluated methods. The developed extractor reduces hand labor, is affordable, and is effective in capturing larvae and adults of H. hampei.

ABSTRACT Coffee is the most appreciated beverage in worldwide; Brazil is the largest producer and exporter of this commodity. Organochlorine endosulfan was banned from the country in 2013 due to its teratogenic agent-related features. Since then, coffee plantations have experienced increased Hypothenemus hampei infestation rates. The aim of the current study is to assess variations in the rates of Coffea arabica fruits brocaded by H. hampei after the application of entomopathogenic fungal species Beauveria bassiana IBCB66. Experiments were carried out with ‘Catuaí’ and ‘Mundo Novo’ cultivars between 2018 and 2020, during the borer transit period. Three experiments were carried out based on the application of the aforementioned fungal species on the investigated coffee plant species, both by spraying and sprinkling, at 30-day intervals; 10 fruits were collected per face of each useful plant in each repetition. The experiment has followed a randomized blocks design with five treatments, including the control, and five repetitions, each. Beauveria bassiana Ecobass (IBCB66) wettable powder spray, at the concentration of 2 × 1013·ha–1, was used in experiments I and II. On the other hand, the mix used in experiment III was prepared with blastospores at concentration of 5 × 1012·ha–1 blastospores + 0.1% Silwet. The sprinkling process in all three experiments has used dry aerial conidia at concentration of 2 × 1013·ha–1. Collected data were subjected to analysis of variance (ANOVA), which was followed by Fisher’s test at 5% probability level, in the SISVAR software. More than 35,000 fruits were assessed. In addition to variations between experiments, results have evidenced that the rate of brocaded fruits remained high.

Resumen La broca del café es un organismo poiquilotermo, cuyo desarrollo y tamaño poblacional dependen proporcionalmente de la temperatura. Consecuentemente, las pérdidas económicas ocasionadas por esta plaga en el cultivo tienen una relación directa con la temperatura y otras condiciones ambientales. Objetivo. Evaluar el efecto del gradiente altitudinal y de diferentes sistemas de sombrío en cafetales sobre la infestación de broca del café. Metodología. Se evaluó el porcentaje de infestación en doce fincas, en un rango altitudinal comprendido entre 1430 y 2030 m y en diferentes niveles de sombrío, incluyendo cafetales a plena explosión solar. En cada localidad se monitorearon las condiciones ambientales por medio de estaciones climáticas. Resultados. Se encontró que los porcentajes de infestación están inversamente relacionados con la altitud, que, a su vez, está directamente influenciada por la temperatura. Por otro lado, hubo una interacción significativa entre la altura y el sombrío, sugiriendo así que los cafetales con sombrío presentaron mayores porcentajes de infestación. Sin embargo, la mayor incidencia de broca se evidenció en cafetales por debajo de 1600 msnm con mayores niveles de sombrío. Adicionalmente se verificó que el aumento de sombrío está inversamente relacionado con la radiación fotosintéticamente activa, factor que potencialmente influenciaría la humedad de los frutos, afectando así indirectamente el desarrollo de la broca del café. Finalmente se calculó el número de generaciones mensuales de cada localidad evaluado, obteniendo 0.3 generaciones en localidades con temperaturas promedio de 17 °C y de una generación al mes en localidades con temperaturas promedio de 23 °C. Conclusión. El número de generaciones y el tamaño poblacional de la broca del café, se ven influenciados drásticamente por condiciones ambientales como la elevación y el sombrío del cultivo de café. Por lo tanto, estas son condiciones que se deben tener en cuenta en la planeación del manejo de esta plaga de gran importancia para la producción de café.

Abstract Coffee berry borer is a poikilothermic organism whose development and population size depend proportionally on temperature. Consequently, the economic losses caused by this pest in the crop have a direct relationship with the temperature and other environmental conditions. Objective: To evaluate the effect of the altitudinal gradient and of different shade systems in coffee plantations on the infestation of coffee berry borer. Methodology: The infestation percentage was evaluated in twelve farms in an altitudinal range between 1430 and 2030 m at different levels of shade, including coffee plantations at full solar explosion. In each locality, environmental conditions were monitored through weather stations. Results: Infestation percentages were found to be inversely related to altitude, which, in turn, are directly influenced by temperature. On the other hand, there was a significant interaction between height and shade, thus suggesting that coffee plantations with shade had higher percentages of infestation. However, the highest incidence of coffee berry borer was evidenced in coffee plantations below 1600 meters above sea level with higher levels of shade. Additionally, it was verified that the increase in shade is inversely related to photosynthetically active radiation, a factor that could potentially influence the humidity of the fruits, thus indirectly affecting the development of coffee borer. Finally, the number of monthly generations of each evaluated locality was calculated, obtaining 0.3 generations in localities with average temperatures of 17 °C and one generation per month in localities with average temperatures of 23 °C. Conclusion: The number of generations and the population size of coffee berry borer is drastically influenced by environmental conditions such as the elevation and shade of the coffee crop. Therefore, these are conditions that must be considered when planning the management of this pest of great importance for coffee production.

Abstract The coffee borer beetle Hypothenemus hampei is one of the most important pest insects worldwide that cause damage to coffee berries. The objective was to evaluate vinegars and plant extracts mixed with ethanol to be used as attractants for the capture of coffee borer beetles. The following mixtures were used for the first time: ethanol-basil, ethanol-rue, ethanol-rosemary, ethanol-white vinegar, and ethanol-apple vinegar, in a proportion of 1:1 and as a control, the methanol-ethanol mixture 3:1 using handmade traps in a completely randomized design with five repetitions per treatment. Based on the statistical analyses, we observed that ethanol-apple vinegar and ethanol-white vinegar resulted in the highest capture of specimens, with a mean of 136.36 and 98.24, respectively, compared to the other mixtures. In conclusion, the most efficient attractants in this study could be used as a sustainable strategy for the capture of coffee borer beetles.

Resumen La broca del café Hypothenemus hampei, es uno de los insectos plaga más importantes a nivel mundial que dañan al fruto del café. El objetivo fue evaluar vinagres y extractos de plantas mezclados con etanol para ser utilizados como atrayentes para la captura del insecto. Se utilizaron por primera vez las mezclas: etanol-albahaca, etanol-ruda, etanol-romero, etanol- vinagre blanco y etanol-vinagre de manzana, en una proporción 1:1 y como testigo metanol-etanol 3:1, utilizando trampas artesanales, en un diseño completamente al azar con cinco repeticiones por cada tratamiento. Con base en los análisis estadísticos, se observó que el etanol-vinagre de manzana y etanol-vinagre blanco, presentaron la mayor captura de especímenes 136,36 y 98,24 en promedio con respecto a las otras mezclas. Se concluye que los mejores atrayentes utilizados en esta investigación pueden servir como una estrategia sustentable para la captura de la broca del café.

Resumo: O objetivo deste trabalho foi avaliar a atividade inseticida de formulação botânica com óleo de nim (Azadirachta indica) e D-limoneno no controle de Hypothenemus hampei, bem como determinar a composição de ácidos graxos do óleo de nim. O óleo de nim 95% foi extraído de sementes de A. indica prensadas a frio, e o D-limoneno, da casca de frutas cítricas. Extrato húmico (carbono orgânico), potássio, cálcio, magnésio e enxofre foram adicionados como aditivos. A análise do óleo de nim por cromatografia gasosa acoplada à espectrometria de massa mostrou que o ácido linoleico (49,28%) é o principal ácido graxo na sua composição. Os testes em campo foram realizados em cafeeiro (Coffea arabica) 'Topázio MG 1190', com quatro aplicações do composto formulado, a cada 20 dias, por um período de 64 dias. Para análise da atividade inseticida, a formulação botânica foi aplicada em papel filtro ou topicamente no dorso do inseto, tendo mostrado mortalidade de 63,34 e 100% após 48 horas, respectivamente. Em campo, a atividade inseticida causou redução de 62,4% na incidência da população de H. hampei, em comparação ao controle. A formulação botânica avaliada é eficiente no controle de H. hampei em cafeeiro.

Abstract: The objective of this work was to evaluate the insecticidal activity of a botanical formulation with neem (Azadirachta indica) oil and D-limonene for the control of Hypothenemus hampei, as well as to determine the fatty acid composition of neem oil. Ninety-five percent neem oil was extracted from cold-pressed A. indica seeds, and D-limonene, from citrus peel. Humic extract (organic carbon), potassium, calcium, magnesium, and sulfur were included as additives. The analysis of neem oil by gas chromatography coupled to mass spectrometry showed that linoleic acid (49.28%) is the main fatty acid in its composition. Field tests were carried out on 'Topázio MG 1190' coffee (Coffea arabica) trees, with four applications of the formulated compound every 20 days for a period of 64 days. For the evaluation of insecticidal activity, the botanical formulation was applied to filter paper or topically on the insect’s dorsal side, showing a mortality of 63.34 and 100% after 48 hours, respectively. In the field, insecticidal activity caused a 62.4% reduction in the incidence of the H. hampei population, when compared with the control. The evaluated botanical formulation is efficient in controlling H. hampei in coffee plants.

RESUMO A broca-do-café, Hypothenemus hampei, é a principal praga do cafeeiro na Amazônia e em todo mundo. Embora haja cultivo de café no estado do Amazonas, não há informações sobre o comportamento da população desta espécie nesta região do Brasil. Como a região amazônica é um ambiente propício para o H. hampei, o presente trabalho objetivou investigar a população do inseto em diferentes sistemas de cultivo implantadas no município de Humaitá, Sul do Amazonas: i) café em floresta nativa (CFN), ii) café consorciada com cupuaçu e açaí (CCA) e iii) café em monocultivo (MC). Também, utilizou-se os parâmetros climáticos e fenológico da planta para relacionar com a dinâmica populacional do inseto. Para isso, instalaram-se 10 armadilhas iscadas em cada lavoura para amostragens mensais da população do H. hampei durante Julho/2018 a Junho/2019. A maior abundância populacional do inseto foi observada durante a floração do café, principalmente na lavoura de monocultivo, em relação às demais lavouras consorciadas. O valor médio total da população do inseto observada nos três ambientes investigados é maior no mês de agosto/2018, registrando-se o valor de 1749. O aumento dos índices pluviométricos ocorreu simultaneamente com a diminuição da população inseto, isso coincide com o final da floração (outubro/2018, 161.8 mm) e início da granação dos frutos (novembro/2018, 234.8 mm). A menor abundância do inseto observada na lavoura sob floresta nativa, abundância de 83 (CFN), indica que a biodiversidade local desfavorece o tamanho populacional da praga no ambiente. As fases fenológicas do café conilon e a influência das condições climáticas podem resultar em mudanças na dinâmica populacional de H. hampei. Os cafeeiros em sistemas agroflorestais são mais promissores para manter essa população em níveis baixos.

ABSTRACT Hypothenemus hampei is the main coffee pest in the Amazon and worldwide. Although coffee cultivation exists in the State of Amazonas, there is no information regarding the behavior of this species population in this region of Brazil. Since the Amazon region is a favorable environment for H. hampei, the present study aimed to investigate this insect population in the following cultivation systems implanted in the municipality of Humaitá, Southern Amazonas: i) coffee in native forest (CNF), ii) coffee intercropped with cupuaçu and açaí (native Brazilian fruits) (CCA) and iii) coffee in monoculture (MC). Also, the climatic and phenological parameters of the plant were used in relation with the population dynamics of the insect. For that, ten baited traps were installed in each field for the monthly sampling of the H. hampei population from July/2018 to June/2019. The highest population abundance of the insect was observed during the coffee flowering season, mainly in monoculture crops, when compared to the other intercropped fields. The total mean value of the insect population observed in the three environments evaluated was highest in August/2018, with a recorded value of 1749. The increase in rainfall indexes occurred simultaneously with a decrease in the insect population; this coincides with the end of the flowering season (October/2018, 161.8 mm) and the beginning of grain formation (November/2018, 234.8 mm). The lowest abundance of the insect observed in the coffee fields in native forest, with an abundance of 83 (CNF), indicates that the local biodiversity disfavors the population size of the pest in the environment. These results suggest that phenological phases of conilon coffee and the influence of climatic conditions can result in changes in the population dynamics of the H. hampei. Coffee agroforestry systems are more promising to maintain this population at low levels.

Resumen Introducción. La broca del café (Hypothenemus hampei, CBB) es una de las plagas más devastadoras en plantaciones de café alrededor del mundo. Aunque CBB es susceptible al efecto de las δ-endotoxinas del Bacillus thuringiensis subs. israelensis (Bti) a nivel de laboratorio, la eficacia de este método de control es deficiente en campo, posiblemente debido a la inactivación ocasionada por proteasas digestivas diferentes a las requeridas para la activación de las protoxinas. Objetivo. Determinar si la incorporación de un extracto de harina de soya con inhibidores de proteasas (PI) mezclado con cristales y esporas de Bti (Bti-sc) en una dieta artifical, podría mejorar la toxicidad de Bti contra CBB. Materiales y métodos. Este estudio se realizó en San José, Costa Rica entre 2012 y 2013. Se expuso un conjunto de insectos hembras adultas de CBB a una mezcla que incluyó diferentes concentraciones del extracto de soya parcialmente purificado con PI activo y un liofilizado de Bti-sc, y fueron evaluadas mediante un bioensayo en dieta artificial para estimar la concentración subletal (CL50). Los resultados de mortalidad se validaron mediante observación del intestino medio diseccionado, cuya ultraestructura se analizó mediante microscopía electrónica de transmisión. Resultados. El extracto de soya degradó parcialmente el complejo Bti-sc, redujo la CL50 en casi cuatro veces (de 1,135 a 0,315 µg µl-1) y potenció la mortalidad de CBB de manera concentración-dependiente. Los análisis histológicos del intestino medio confirmaron este efecto sinérgico, dado que se visualizaron daños severos en el epitelio intestinal de CBB expuestos a Bti-sc + PI comparado con Bti-sc solo. Conclusiones. La combinación del extracto de soya enriquecido con PI y Bti-sc potenció la letalidad sobre CBB, que se confirmó por el colapso intestinal. La harina de soya es un suplemento económico que podría aumentar la efectividad de Bti para controlar CBB y retrasar el desarrollo de resistencia biológica.

Abstract Introduction. The coffee berry borer (Hypothenemus hampei Ferrari, CBB) is one of the most devastating pests on coffee plantations around the world. Although CBB is susceptible to the effect of δ–endotoxins of Bacillus thuringiensis subs. israelensis (Bti) at laboratory level, the efficacy of this control method is poor in the field, presumably due to the inactivation by digestive proteases different to those required for protoxin activation. Objective. To study whether the addition of a soybean flour extract enriched with protease inhibitors (PI), mixed with Bti crystals and spores (Bti-sc) in an artificial diet, could improve the toxicity of Bti against CBB. Materials and methods. This study was performed in San José, Costa Rica, between 2012 and 2013. A set of adult female CBB insects was exposed to a mixture containing different concentrations of a partially purified soybean meal extract with active PI and lyophilized Bti-sc, and were tested through a bioassay in artificial diet to estimate the sub-lethal concentration (LC50). The mortality results were validated by observing the dissected midgut, whose ultrastructure was analyzed by transmission electron microscopy. Results. The soybean extracts partially degraded the Bti-sc complex, it reduced its LC50 by almost four times (from 1.135 to 0.315 µg µl-1) and enhanced CBB mortality in a concentration-dependent manner. Histological analyses of the midgut confirmed this synergistic effect, since severe epithelial damage to the intestinal epithelium of CBB exposed to Bti-sc + PI was visualized compared to Bti-sc alone. Conclusions. The combination of a soybean extract enriched in PI and Bti-sc enhanced the mortality effect over CBB, which was confirmed by the midgut collapse. Soybean flour is a cost-effective supplement that could increase Bti effectiveness against CBB and delay the appearance of biological resistance.

ABSTRACT Coffee berry borer (Hypothenemus hampei), is one of the main pests affecting coffee production worldwide. This pest feeds and reproduces inside the berry reducing its weight and quality, causing losses up to 50%. Some strains of Bacillus have plasmids that incorporate cry and cyt genes, whose products have toxic properties to insects. Bacillus thuringiensis and Bacillus pumilus have been evaluated as entomopathogenic. The objectives of this study was to determine the potential of two native Bacillus strains (B. thuringiensis Bt3971 and B. pumilus Bp4185) isolated from coffee ecosystem of Puerto Rico as biocontrol of coffee berry borer. The presence of a 12 kbp plasmid was confirmed in Bt3971 and Bp4185, and a 116 kDa protein with entomopathogenic potential from Bt3971. After 48 h of treatment, Bt3971 (A590=0.8, approximately 1010 CFU ml-1) caused 55% mortality and 82% at 72 h in a population of insects. Bp4185 strain, at 72 h caused 55% mortality (A590=1.0, approximately 1012 CFU ml-1) and 46% at 96 h (A590=0.8). Both native bacterial strains, adapted to the environmental conditions of Puerto Rico, have potential for biological control of coffee berry borer.

RESUMEN La broca del café (Hypothenemus hampei) es un de las plagas principales del cultivo y efecta su producción a nivel mundial. El insecto crece y se reproduce dentro de la fruta y reduce su peso y calidad, lo que causa pérdidas hasta del 50%. Algunas cepas del género Bacillus contienen plásmidos con genes del tipo cry y cyt que codifican por proteínas tóxicas a insectos. Bacillus thuringiensis y Bacillus pumilus se han evaluado como entomopatógenas. Este estudio tuvo como objetivo determinar el potencial de Bacillus thuringiensis Bt3971 y Bacillus pumilus Bp4185 para el biocontrol de H. hampei. La presencia de plásmidos de 12 kbp fue confirmada en ambas cepas, así como una proteína de 116 kDa potencialmente entomopatógena en Bt3971. Después de 48 h de tratamiento, Bt3971 ocasionó 55% de mortalidad (A590=0.8, aproximadamente 1010 UFC ml-1) y 82% a las 72 h en la población de insectos. Bp4185 a las 72 h produjo 55% de mortalidad (A590=1.0, aproximadamente 1012 UFC ml-1) y 46% a las 96 horas (A590=0.8). Ambas cepas nativas, adaptadas a las condiciones ambientales de Puerto Rico, tienen potencial para el control biológico de la broca del café.

Abstract Coffee is one of the most relevant crops in Mexico, its importance lies in the fact that it is a sustainable crop, it has been adopted and adapted by indigenous communities and generates foreign exchange for the country. One of the most important coffee growing regions in the country is the South Pacific, made up of Chiapas, Oaxaca and Guerrero where 53% of the coffee is produced and 60% of the producers of this aromatic are found. The presence of pests and diseases in coffee can generate losses between 30 to 50% of production. The Hypothenemus hampei (Ferrari) coffee berry borer is the most serious entomological problem of this crop and its presence in the country is over 40 years old, currently it is found in all states that grow coffee. That is why, in this work, a practical methodology was generated that spatially calculated the degrees days of development of the coffee berry borer through the surface temperature recorded by Modis images with a spatial resolution of 1 km and with a one-day temporary resolution, monitoring from April 01, 2018 to March 31, 2019. The results indicated that up to 13 generations of the pest could form in this region, presenting risk almost all year round, both due to development and pest spread. This methodology can help strengthen the monitoring of the pest in the country.

Resumen El café es uno de los cultivos más relevantes en México, su importancia radica en que es un cultivo sustentable, ha sido adoptado y adaptado por comunidades indígenas y genera divisas para el país. Una de las regiones caficultoras más importantes del país, es la del Pacífico Sur, compuesta por Chiapas, Oaxaca y Guerrero donde se produce 53% del café y se encuentra 60% de los productores de este aromático. La presencia de las plagas y enfermedades en el café puede generar pérdidas entre 30 al 50% de la producción. La broca de café Hypothenemus hampei (Ferrari) es el problema entomológico mas grave de este cultivo y su presencia en el país es de hace más de 40 años, actualmente se encuentra en todos los estados que cultivan café. Es por ello, que en este trabajo se generó una metodología práctica que calculó espacialmente los grados días de desarrollo de la broca de café a través de la temperatura de la superficie registrada por imágenes Modis con una resolución espacial de 1 km y con una resolución temporal de un día, monitoreando del 01 de abril de 2018 al 31 de marzo de 2019. Los resultados indicaron que en esta región se puede formar hasta 13 generaciones de la plaga, presentando riesgo en casi todo el año, tanto por el desarrollo como en la dispersión de la plaga. Esta metodología puede apoyar a fortalecer el monitoreo que se hace de la plaga en el país.

Resumen En el estado de Tabasco, México, la broca del café Hypothenemus hampei, se registró por primera vez en 2004. Desde entonces se ha convertido en la principal limitante insectil de la producción de café. El objetivo del presente trabajo fue identificar la micobiota asociada a adultos de H. hampei, recolectados en plantaciones de café en tres municipios de Tabasco: Teapa, Tacotalpa y Huimanguillo. Se recolectaron cerezas infestadas con la broca y se extrajeron los adultos de H. hampei. Los hongos que crecieron sobre los cadáveres de este Scolytinae se aislaron e identificaron morfológicamente. Dos aislamientos de cada especie morfológica se identificaron molecularmente. Se obtuvo un total de 74 aislamientos de la cutícula de H. hampei, de los cuales, 52 presentaron afinidad morfológica con Fusarium solani, cinco con F. oxysporum, cinco con F. verticillioides, uno con Geotrichum candidum y 11 con Beauveria bassiana. Las secuencias de ADN mostraron similitud con secuencias del GenBank, con homologías de 99 y 100 %, las cuales coincidieron con cada especie morfológica. La especie más abundante fue F. solani (70,2 %), seguida de B. bassianna (14,8 %). Las especies F. verticillioides y F. oxysporum representaron cada una el 6,7 %. La menos abundante fue G. candidum (1,3 %). Los hallazgos de F. verticillioides y G. candidum son nuevos registros de hongos micromicetos asociados a H. hampei.

Abstract In the state of Tabasco, Mexico, the coffee berry borer Hypothenemus hampei, was reported for the first time in 2004. Since then it has become the main insect pest of coffee in this area. The aim of the present study was to document the mycobiota associated to H. hampei adults in coffee plantations of three localities of Tabasco: Teapa, Tacotalpa and Huimanguillo. Coffee berries infested with the coffee berry borer were collected from the field and adults of H. hampei were extracted. The fungi that grew on the Scolytinae cadavers were isolated and identified using taxonomic keys. Two isolates of each morphological species were molecularly identified. A total of 74 isolates was obtained from the cuticle of H. hampei. Of them, 52 presented morphological affinity with Fusarium solani, five with F. oxysporum, five with F. verticillioides, one with Geotrichum candidum and 11 with Beauveria bassiana. The DNA sequences showed similarity with sequences found in GenBank, with homologies of 99 % and 100 %, which coincided with each morphological species. The most abundant species was F. solani (70.2 %), followed by B. bassiana (14.8 %). The species F. verticillioides and F. oxysporum represented each one, at 6.7 %, respectively. The least abundant was G. candidum (1.3 %). The findings of the species F. verticillioides and G. candidum are new reports of micromycetes fungi associated with H. hampei. This study contributes to the knowledge of the diversity of fungi associated with H. hampei.