Feeding Preferences of the Bean Leaf Beetle (Ootheca spp.) (Coleoptera: Chrysomelidae): Insights for Targeted Pest Control Strategies in Uganda.

The bean leaf beetle (BLB) (Ootheca spp.) is a polyphagous pest causing significant yield losses in Uganda, particularly in the Northern and Eastern regions on various hosts plants. Despite its polyphagous behaviour, the BLB exhibits preferential feeding, offering an opportunity for targeted pest management. This study explored its feeding preferences across seven crops: common bean, cowpea, greengram, okra, roselle (malakwang), groundnuts, and soybean. This study was conducted in Arua and Lira districts using a randomized complete block design for two rainy seasons (2018A and 2018B). The results showed significant differences in BLB abundance and foliar damage among host crops, locations, days after planting and seasons. Cowpea was the most preferred crop while groundnuts was the least preferred. Therefore, cowpea can be recommended for use as a trap for managing Ootheca spp. in gardens where it is not the main crop. There was a higher pest abundance in Arua than in Lira. There was also a higher pest abundance in 2018A than in 2018B. These findings highlight the importance of understanding BLB's feeding preferences for implementing effective IPM strategies, emphasizing the potential role of trap cropping, especially with cowpea, to minimize BLB damage in resource-constrained agricultural settings.

Host and Seasonal Effects on the Abundance of Bean Leaf Beetles (Ootheca spp.) (Coleoptera: Chrysomelidae) in Northern Uganda.

Bean leaf beetles (BLBs) (Ootheca spp.) are serious legume pests in Uganda and sub-Saharan Africa, but their ecology is not well understood. We planted host plants, viz., common bean, cowpea, and soybean, in an experiment in the hotspot areas of Arua and Lira districts in Northern Uganda in order to assess their influence on the density of adults and immature stages of BLBs in different seasons. Overall, the number of adults, larvae, and pupae were higher in cowpea than common bean and soybean plots. The number of adults were highest in cowpea (29.5 adults/15 plants) in Arua during the long rainy season (2018A). The number of adults did not differ significantly during short rains (season B) in 2017 and 2018. Similarly, in Lira district, the highest number of adult BLBs was in cowpea (4.6 beetles) compared to the common bean (2.7 beetles) and soybean plots, with a peak at four weeks after planting (WAP). During 2018A, larvae of BLBs first appeared at five WAP and seven WAP and peaked at 13 WAP and 11 WAP in Arua and Lira, respectively. The pupae were present in the soil after the harvesting of crops during 2018A, but peaked at seven WAP and eight WAP in 2018B season in Arua and Lira, respectively. The occurrence of below-ground adults in 2018B followed the peak abundance of pupae, although this was delayed until six WAP in Arua compared to Lira. We conclude that cowpea is the most preferred by adults and larvae compared to common bean and soybean. Similarly, the first rain season (2018A) attracted higher abundance and damage than the second rain season. Management of the BLBs should thus take into consideration avoidance of host crop rotation and dealing with the below-ground stages.

Bean Leaf Beetle (Ootheca spp.) (Coleoptera: Chrysomelidae) Management via Planting Timing and Insecticides.

Bean leaf beetles (Ootheca spp.) (Coleoptera: Chrysomelidae) are one of Africa's most important pests of the common bean (Phaseolus vulgaris L.). Roots, leaves, floral parts, and young pods are all attacked, leading to a considerable loss in grain yield. In Uganda, there are no comprehensive prescribed management strategies for bean leaf beetles, but farmers typically try to control the pest by delaying bean crop sowing, and to a lesser extent, using insecticides. Although farmers have consistently implemented the two approaches, there is no information on the effects of the approaches in Uganda. To assess the impact of planting timing and insecticide spray regimes on bean leaf beetle populations, concomitant foliar damage, and grain yield, we set up trials in three agro-ecological zones with known presence of the beetles during the second rainy season of 2016 (2016) and the first rainy season of 2017 (2017). The first planting, coinciding with early planting, was conducted within one week after the onset of rains. The second planting, coinciding with mid planting, followed two weeks later, while the third planting, considered late planting in this study, was conducted one month after the second planting. A foliar application of cypermethrin commencing at 7 days after emergence (DAE), 14 DAE, 21 DAE, 28 DAE, and 35 DAE; a soil drench of imidacloprid at planting combined with a foliar spray starting at 7 DAE; and an untreated control were among the insecticide spray regimes evaluated. Higher bean leaf beetle abundance was recorded from mid-planting, while higher foliar damage was recorded from late planting in two of the three agro-ecological zones. However, higher marketable grain yield was recorded from early planting in all agro-ecological zones, suggesting that delayed planting may not be beneficial. Insecticide application reduced foliar damage and increased marketable grain yield, with a combination of soil drench and foliar spray resulting in much less foliar damage and, as a result, higher grain yield. However, this did not result in economic benefits. Furthermore, marketable grain yield was higher when insecticide spray regimes were combined with early planting in all agro-ecological zones during both seasons. Our findings suggest that the common bean should be planted early and that the control of the bean leaf beetle should target both the adults and the juvenile stages in the soil. Therefore, there is a need for farmers to be able to access less-expensive soil treatments.

Host Plant-Based Artificial Diets Enhance Development, Survival and Fecundity of the Edible Long-Horned Grasshopper Ruspolia differens (Orthoptera: Tettigoniidae).

Wild swarms of the long-horned grasshoppers Ruspolia differens (Serville) which are widely harvested for consumption and sale in Africa are seasonal and unsustainable, hence the need for innovative ways of artificially producing the insects. We investigated the development, survival, and reproduction of R. differens in the laboratory on diets mixed with host plants [Digitaria gayana Kunth, Cynodon dactylon (L.) and Megathyrsus maximus Jacq (Poales: Poaceae); Ageratum conyzoides L. (Asterales: Asteraceae)] identified from guts of their wild conspecifics with a view to developing a suitable diet for artificial mass rearing of the edible insect. A standard diet comprising ground black soldier fly, Hermetia illucens L. (Diptera: Startiomyidae) larvae, soybean flour, maize flour, vitamin premix, and ground bones was tested for rearing R. differens as a control against the same ingredients incorporated with individual powders of the different host plants. Whereas R. differens developed more slowly in the diet mixed with D. gayana than in the control diet; its development was faster in the diet mixed with C. dactylon. Mortalities of R. differens in host plant-based diets were 42.5-52.5%, far lower than in the control diet with 71% mortality. The insects raised on the diet mixed with M. maximus laid approximately twice more eggs compared to R. differens fecundities from the rest of the diets. However, inclusion of host plants in the diets had no detectable influence on R. differens adult weight and longevity. These findings support inclusion of specific host plants in artificial diets used for mass rearing of R. differens to enhance its survival, development, and fecundity.

Distribution and Relative Abundance of Bean Leaf Beetles (Ootheca spp.) (Insecta: Coleoptera: Chrysomelidae) in Uganda.

Bean leaf beetles (Ootheca spp.) (Insecta: Coleoptera: Chrysomelidae) are one of Africa's most destructive pests of common bean and other leguminous crops. The beetles are widely distributed in Africa where they are estimated to cause annual crop yield losses of 116,400 tons of crop yields in sub-Saharan Africa. Despite their importance, little is known about the distribution, relative abundance and damage caused by bean leaf beetles in Uganda. As a result, the development of effective management methods has been hampered. We conducted surveys in six key Ugandan agro-ecological zones to determine the species distribution and relative abundance of bean leaf beetles. Findings indicate that leaf beetles belonging to 12 genera are present, including members of the genera Afrophthalma Medvedev, 1980, Buphonella Jacoby, 1903, Chrysochrus Chevrolat in Dejean, 1836, Diacantha Dejean, 1845, Exosoma Jacoby, 1903, Lamprocopa Hincks, 1949, Lema Fabricius, 1798, Nisotra Baly, 1864, Neobarombiella Bolz and Wagner, 2012, Ootheca Dejean, 1935, Parasbecesta Laboissière, 1940, and Plagiodera Dejean, 1835. We identified only three species belonging to the genus Ootheca: O. mutabilis, O. proteus, and O. orientalis. Seventy percent of all the beetles collected were O. mutabilis and these were present in all agro-ecological zones studied. The Northern Moist Farmlands (21.9%), West Nile Farmlands (12.9%), Central Wooded Savanna (4.4%) and Southern and Eastern Lake Kyoga Basin (1.4%) were the only agro-ecological zones where O. proteus was found. Only one specimen of O. orientalis was found at a single site in the Central Wooded Savanna. The Northern Moist Farmlands had a significantly (p < 0.05) higher bean leaf beetle density than the West Nile Farmlands and Southwestern Highlands. Similarly, the Northern Moist Farmlands had the highest beetle foliar damage per plant (1.15 ± 0.05), while the Southwestern Highlands had the lowest (0.03 ± 0.02). We provide the first information on Ootheca species distribution, abundance and damage in Uganda. Our findings provide a foundation for assessing the importance of Ootheca spp. as common bean pests in Uganda.

Can Occurrence and Distribution of Ground Beetles (Carabidae) Be Influenced by the Coffee Farming System in the Mount Elgon Region of Uganda?

The Mount Elgon region of Uganda has coffee farmlands distributed along the slopes of the mountain, in a mosaic of differing crop combinations, and semi-natural vegetation. Thus, there are parcels of varying microclimate that create disparities in occurrence of key insect functional groups. The study quantified the occurrence of Carabidae in 72 coffee farmlands categorized by altitude: low (1400-1499 m.a.s.l), mid (1500-1679 m.a.s.l), and high (1680-2100 m.a.s.l); and farming system: coffee monocrop, coffee+annual crops, coffee+banana, and coffee+banana+shade trees. The results revealed highly significant effects of altitude, farming systems, and the interaction of the two on occurrence of three Carabidae genera (Anisodactylus, Chlaenius, and Harpalus.). The abundance of Harpalus spp. was higher at lower altitudes in coffee monocropped farming systems; Anisodactylus spp. were more abundant at higher altitudes in coffee+annual crop systems; and Chlaenius spp. were highest in the coffee+banana+shade tree system at mid altitudes. The belowground microclimate parameters of soil moisture, pH, EC; and the aboveground diversity of semi-natural vegetation explained some of the differences in occurrence of the different Carabidae genera. This distinctiveness in preference of different genera in the same family hinders collective recommendations but looks to a more pragmatic strategy in nurturing diversity on a holistic scale.

Predicting the current and future distribution of the edible long-horned grasshopper Ruspolia differens (Serville) using temperature-dependent phenology models.

The edible long-horned grasshopper Ruspolia differens (Serville) is widely distributed and consumed in sub-Saharan Africa. Efficient mass rearing of the edible grasshopper is critical to ensure their sustainable supply for food and nutritional security. Hence, we investigated the effect of temperature on development, survival and reproduction of R. differens under six constant (15, 20, 25, 30, 32 and 35 °C) and fluctuating temperatures. Using Insect Life Cycle Modeling software we fitted, linear and non-linear models to R. differens development, mortality, longevity, and fecundity. The best-fitted functions were compiled for each life stage to yield a phenology model, which was stochastically simulated to estimate the life table parameters. We used the process-based climatic phenology models, and applied establishment risk index (ERI) and generation index (GI) in a geographic information system to map the potential distribution of R. differens under current and future climates. At optimum temperatures of 30-32 °C, egg incubation period was 14-15 days and the developmental time was shortest at 52.5-58 days. Lowest nymphal mortality (3.4-13%) and the highest female fecundity was obtained at 25-30 °C. The optimum temperature for the reproduction ranged between 27 and 30 °C. Most simulated lifetable parameters were at their maximum at 28 °C. Predictive models showed that countries in the East, Central, West, Southern and the Horn of Africa were suitable for establishment of R. differens under current climate scenarios (2000). However, by 2050, climatically suitable areas for the establishment of R. differens were predicted to shrink in the West, Southern and the Horn of Africa than its current distribution. We predict up to three generations per year for R. differens in sub-Saharan Africa under current scenarios which can increase to 4 under future scenarios. The optimum rearing temperatures identified can guide optimization of mass rearing of R. differens.

Factors Influencing Genomic Prediction Accuracies of Tropical Maize Resistance to Fall Armyworm and Weevils.

Genomic selection (GS) can accelerate variety improvement when training set (TS) size and its relationship with the breeding set (BS) are optimized for prediction accuracies (PAs) of genomic prediction (GP) models. Sixteen GP algorithms were run on phenotypic best linear unbiased predictors (BLUPs) and estimators (BLUEs) of resistance to both fall armyworm (FAW) and maize weevil (MW) in a tropical maize panel. For MW resistance, 37% of the panel was the TS, and the BS was the remainder, whilst for FAW, random-based training sets (RBTS) and pedigree-based training sets (PBTSs) were designed. PAs achieved with BLUPs varied from 0.66 to 0.82 for MW-resistance traits, and for FAW resistance, 0.694 to 0.714 for RBTS of 37%, and 0.843 to 0.844 for RBTS of 85%, and these were at least two-fold those from BLUEs. For PBTS, FAW resistance PAs were generally higher than those for RBTS, except for one dataset. GP models generally showed similar PAs across individual traits whilst the TS designation was determinant, since a positive correlation (R = 0.92***) between TS size and PAs was observed for RBTS, and for the PBTS, it was negative (R = 0.44**). This study pioneered the use of GS for maize resistance to insect pests in sub-Saharan Africa.

Identification of Edible Short- and Long-Horned Grasshoppers and Their Host Plants in East Africa.

There is a paucity of information on the edible grasshoppers and their host plants in East Africa. This study adopted morphological and molecular analysis to identify edible grasshoppers in Kenya and Uganda. The associated host plants were identified through molecular analysis of the gut contents of the grasshoppers. The cytochrome b and 16s gene primers were used for grasshopper DNA analysis; while matK gene primers were used for plant DNA analysis. All long-horned grasshoppers sampled were identified as Ruspolia differens (Serville) (Orthoptera: Tettigonidae); whereas short-horned grasshoppers were identified as Acanthacris ruficornis (Fabricius) (Orthoptera: Acrididae) and Cyrtacanthacris tatarica (L.) (Orthoptera: Acrididae). Host plants of A. ruficornis were Achyranthes aspera (L.), Centella virgata L.f. Drude, Digitaria gayana (Kunth), Galinsoga quadriradiata Ruiz and Pavon, and Triumfetta pilosa Roth; whereas those of C. tatarica were Alysicarpus rugosus (Willd.) DC and Teramnus uncinatus (L.) SW. Host plants of R. differens were Ageratum conyzoides (L.), Citrus depressa Hayata, Cynodon dactylon (L.), D. gayana, Eragrostis mexicana Hornem, Eucalyptus saligna SM., Indigofera arrecta Hochst. ex A. Rich., Persicaria nepalensis (L.), and Sorghum halepense (L.). Information on the host plants of edible grasshoppers can help in the development of their mass rearing protocols.

An atoxigenic L-strain of Aspergillus flavus (Eurotiales: Trichocomaceae) is pathogenic to the coffee twig borer, Xylosandrus compactus (Coleoptera: Curculionidea: Scolytinae).

This study isolated and evaluated virulence of fungal entomopathogens of Xylosandrus compactus - an important pest of Robusta coffee in Sub-Saharan Africa. A survey was conducted in five farming systems in Uganda to isolate entomopathogens associated with X. compactus. Four fungal isolates were screened for virulence against X. compactus in the laboratory at 1 × 107 conidia ml-1 where an atoxigenic L-strain of A. flavus killed 70%-100% of all stages of X. compactus compared with other unidentified isolates which caused 20%-70% mortalities. The time taken by A. flavus to kill 50% of X. compactus eggs, larvae, pupae and adults in the laboratory was 2-3 days; whereas the other unidentified fungal isolates took 4-7 days. The concentrations of A. flavus that killed 50% of different stages of X. compactus were 5 × 105 , 12 × 105 , 17 × 105 and 30 × 105 conidia ml-1 for larvae, eggs, pupae and adults respectively. A formulation of A. flavus in oil caused higher mortalities of X. compactus larvae, pupae and adults in the field (71%-79%) than its formulation in water (33%-47%). The atoxigenic strain of A. flavus could therefore be developed into a safe biopesticide against X. compactus.

Maize Combined Insect Resistance Genomic Regions and Their Co-localization With Cell Wall Constituents Revealed by Tissue-Specific QTL Meta-Analyses.

Combinatorial insect attacks on maize leaves, stems, and kernels cause significant yield losses and mycotoxin contaminations. Several small effect quantitative trait loci (QTL) control maize resistance to stem borers and storage pests and are correlated with secondary metabolites. However, efficient use of QTL in molecular breeding requires a synthesis of the available resistance information. In this study, separate meta-analyses of QTL of maize response to stem borers and storage pests feeding on leaves, stems, and kernels along with maize cell wall constituents discovered in these tissues generated 24 leaf (LIR), 42 stem (SIR), and 20 kernel (KIR) insect resistance meta-QTL (MQTL) of a diverse genetic and geographical background. Most of these MQTL involved resistance to several insect species, therefore, generating a significant interest for multiple-insect resistance breeding. Some of the LIR MQTL such as LIR4, 17, and 22 involve resistance to European corn borer, sugarcane borer, and southwestern corn borer. Eleven out of the 42 SIR MQTL related to resistance to European corn borer and Mediterranean corn borer. There KIR MQTL, KIR3, 15, and 16 combined resistance to kernel damage by the maize weevil and the Mediterranean corn borer and could be used in breeding to reduce insect-related post-harvest grain yield loss and field to storage mycotoxin contamination. This meta-analysis corroborates the significant role played by cell wall constituents in maize resistance to insect since the majority of the MQTL contain QTL for members of the hydroxycinnamates group such as p-coumaric acid, ferulic acid, and other diferulates and derivates, and fiber components such as acid detergent fiber, neutral detergent fiber, and lignin. Stem insect resistance MQTL display several co-localization between fiber and hydroxycinnamate components corroborating the hypothesis of cross-linking between these components that provide mechanical resistance to insect attacks. Our results highlight the existence of combined-insect resistance genomic regions in maize and set the basis of multiple-pests resistance breeding.

Toxicity of seven Bacillus thuringiensis Cry proteins against Cylas puncticollis and Cylas brunneus (Coleoptera: Brentidae) using a novel artificial diet.

"Sweetpotato weevils" Cylas puncticollis (Boheman) and Cylas brunneus F. (Coleoptera: Brentidae) are the most important biological threat to sweetpotato, Ipomoea batatas L. (Lam), productivity in sub-Saharan Africa. Sweetpotato weevil control is difficult due to their cryptic feeding behavior. Expression of Cylas-active Bacillus thuringiensis (Bt) Cry proteins in sweetpotato could provide an effective control strategy. Unfortunately, Bt Cry proteins with relatively high toxicity against Cylas spp. have not been identified, partly because no published methodology for screening Bt Cry proteins against Cylas spp. in artificial diet exists. Therefore, the initial aim of this study was to develop an artificial diet for conducting bioassays with Cylas spp. and then to determine Bt Cry protein efficacy against C. puncticollis and C. brunneus by using this artificial diet. Five diets varying in their composition were evaluated. The highest survival rates for sweetpotato weevil larvae were observed for diet E that contained the highest amount of sweetpotato powder and supported weevil development from first instar to adulthood, similar to sweetpotato storage roots. Seven coleopteran-active Bt Cry proteins were incorporated into diet E and toxicity data were generated against neonate C. puncticollis and second-instar C. brunneus. All Bt Cry proteins tested had toxicity greater than the untreated control. Cry7Aa1, ET33/34, and Cry3Ca1 had LC50 values below 1 microg/g diet against both species. This study demonstrates the feasibility of using an artificial diet bioassay for screening Bt Cry proteins against sweetpotato weevil larvae and identifies candidate Bt Cry proteins for use in transforming sweetpotato varieties potentially conferring field resistance against these pests.