Odor-Mediated Group Organization and Coordination in the Termite-Raiding Ant Megaponera analis (Mayr).

Visual and olfactory communications are vital for coordinated group hunting in most animals. To hunt for prey, the group-raiding termite specialist ant Megaponera analis, which lacks good vision, must first confirm the presence or absence of conspecific raiders. Here, we show that M. analis uses olfactory cues for intraspecific communication and showed greater preference for conspecific odors over clean air (blank) or odors from its termite prey. Chemical analysis of ant volatiles identified predominantly short-chained hydrocarbons. Electrophysiological analysis revealed differential sensory detection of the odor compounds, which were confirmed in behavioral olfactometric choice assays with odor bouquets collected from major and minor castes and the 2 most dominant volatiles and n-undecane n-tridecane. A comparative analysis of the cuticular hydrocarbon profile with those of the short-chained odor bouquet of different populations shows a high divergence in the long-chained profile and a much-conserved short-chained odor bouquet. This suggests that there is less selection pressure for divergence and individual recognition in the short- than the long-chained odor profiles. We conclude that olfactory communication serves as an alternative to visual or sound communication, especially during group raids in M. analis when ants are not in direct contact with one another.

Lemon Terpenes Influence Behavior of the African Citrus Triozid Trioza erytreae (Hemiptera: Triozidae).

The African citrus triozid, Trioza erytreae Del Guercio (Hemiptera: Triozidae) is one of the primary vectors of the bacterium Candidatus Liberibacter spp. which causes citrus greening, a disease of global economic importance in citrus production. Despite its economic importance, little is known about its chemical ecology. Here, we used behavioral assays and chemical analysis to study the chemical basis of interaction between T. erytreae and one of its preferred host plants, Citrus jambhiri. In dual choice Y-tube olfactometer assays, lemon leaf odors attracted females but not males compared to plain air or solvent controls. However, in a petri dish arena assay, both sexes were arrested by lemon leaf odors. Coupled gas chromatography-mass spectrometry (GC/MS) analysis revealed quantitative differences in the odors of flushing and mature leaves, dominated by terpenes. Twenty-six terpenes were identified and quantified. In Petri dish arena assays, synthetic blends of the most abundant terpenes mimicking lemon flushing leaf odors elicited varying behavioral responses from both sexes of T. erytreae. A nine-component blend and a blend of the three most abundant terpenes; limonene, sabinene and ß-ocimene arrested both sexes of T. erytreae. In contrast, a six-component blend lacking in these three components elicited an avoidance response in both sexes. Furthermore, both sexes of T. erytreae preferred the three-component synthetic blend to lemon crude volatile extract. These results suggest that lemon terpenes might be used in the management of T. erytreae.

Low fertility, fecundity and numbers of mated female offspring explain the lower reproductive success of the parasitic mite Varroa destructor in African honeybees.

Although Varroa destructor is the most serious ecto-parasite to the honeybee, Apis mellifera L., some honeybee populations such as Apis mellifera scutellata in Kenya can survive mite infestations without treatment. Previously, we reported that grooming behaviour could be a potential tolerant mechanism expressed by this honeybee subspecies towards mite infestation. However, both hygienic and grooming behaviours could not explain the lower mite-infestation levels recorded in these colonies. Here, we investigated the involvement of other potential resistant mechanisms including suppression of mite reproduction in worker brood cells of A. m. scutellata to explain the low mite numbers in their colonies. High infertility rates (26-27%) and percentages of unmated female offspring (39-58%) as well as low fecundity (1.7-2.2, average offspring produced) were identified as key parameters that seem to interact with one another during different seasons to suppress mite reproduction in A. m. scutellata colonies. We also identified offspring mortality in both sexes and absence of male offspring as key factors accounting for the low numbers of mated daughter mites produced in A. m. scutellata colonies. These results suggest that reduced mite reproductive success could explain the slow mite population growth in A. m. scutellata colonies.

Hygienic and grooming behaviors in African and European honeybees-New damage categories in Varroa destructor.

Varroa destructor is an ectoparasitic pest of honeybees, and a threat to the survival of the apiculture industry. Several studies have shown that unlike European honeybees, African honeybee populations appear to be minimally affected when attacked by this mite. However, little is known about the underlying drivers contributing to survival of African honeybee populations against the mite. We hypothesized that resistant behavioral defenses are responsible for the survival of African honeybees against the ectoparasite. We tested this hypothesis by comparing grooming and hygienic behaviors in the African savannah honeybee Apis mellifera scutellata in Kenya and A. mellifera hybrids of European origin in Florida, USA against the mite. Grooming behavior was assessed by determining adult mite infestation levels, daily mite fall per colony and percentage mite damage (as an indicator of adult grooming rate), while hygienic behavior was assessed by determining the brood removal rate after freeze killing a section of the brood. Our results identified two additional undescribed damaged mite categories along with the six previously known damage categories associated with the grooming behavior of both honeybee subspecies. Adult mite infestation level was approximately three-fold higher in A. mellifera hybrids of European origin than in A. m. scutellata, however, brood removal rate, adult grooming rate and daily natural mite fall were similar in both honeybee subspecies. Unlike A. mellifera hybrids of European origin, adult grooming rate and brood removal rate did not correlate with mite infestation levels on adult worker honeybee of A. m. scutellata though they were more aggressive towards the mites than their European counterparts. Our results provide valuable insights into the tolerance mechanisms that contribute to the survival of A. m. scutellata against the mite.

Vulnerable Habitats Alter African Meliponine Bee's (Hymenoptera: Apidae) Assemblages in an Eastern Afromontane Biodiversity Hotspot.

Habitat degradation has over time formed synergy with other factors to contribute to dwindling populations of both fauna and flora by altering their habitats. The disturbance of natural habitats affects the diversity of both vertebrates and invertebrates by altering both feeding and nesting sites for which organisms are known to depend on for survival. Little is known of the extent to which vulnerable habitats could shape the diversity of most indigent pollinators such as African meliponine bee species in tropical ecosystems. This study was conducted to determine how disturbance could shape the natural occurrence of African meliponine bee species in different ecological habitats of Taita Hills, leading to changes in their diversity. A total of four species depicted by the Renyi diversity profile was recorded in five of the six main habitat types surveyed, and a further extrapolation with Shannon index (EH) also predicted the highest species richness of 4.24 in a deciduous habitat type. These meliponine bee species (Hypotrigona gribodoi, Hypotrigona ruspolii, Meliponula ferruginea (black), and Plebeina hildebrandti) were observed to be unevenly distributed across all habitats, further indicating that mixed deciduous habitat was more diverse than acacia-dominated bush lands, grasslands, and exotic forest patches. Geometric morphometrics categorized all four meliponine bee species into two major clusters-cluster 1 (H gribodoi, H ruspolii, M ferruginea (black)) and cluster 2 (P hildebrandti)-and further discriminated populations against the 4 potential habitats they are likely to persist or survive in. Each habitat appeared to consist of a cluster of subpopulations and may possibly reveal ecotypes within the four meliponine populations. This has revealed that unprecedented conversions of natural habitats to agroecosystems are a key driving factor causing increased habitat isolation and vulnerability in this Afromontane region which may potentially distort local assemblages of native pollinators, such as meliponine bee species.

The Biology and Control of the Greater Wax Moth, Galleria mellonella.

The greater wax moth, Galleria mellonella Linnaeus, is a ubiquitous pest of the honeybee, Apis mellifera Linnaeus, and Apis cerana Fabricius. The greater wax moth larvae burrow into the edge of unsealed cells with pollen, bee brood, and honey through to the midrib of honeybee comb. Burrowing larvae leave behind masses of webs which causes galleriasis and later absconding of colonies. The damage caused by G. mellonella larvae is severe in tropical and sub-tropical regions, and is believed to be one of the contributing factors to the decline in both feral and wild honeybee populations. Previously, the pest was considered a nuisance in honeybee colonies, therefore, most studies have focused on the pest as a model for in vivo studies of toxicology and pathogenicity. It is currently widespread, especially in Africa, and the potential of transmitting honeybee viruses has raised legitimate concern, thus, there is need for more studies to find sustainable integrated management strategies. However, our knowledge of this pest is limited. This review provides an overview of the current knowledge on the biology, distribution, economic damage, and management options. In addition, we provide prospects that need consideration for better understanding and management of the pest.

Behavioral Evidence for Olfactory-Based Location of Honeybee Colonies by the Scarab Oplostomus haroldi.

The Afro-tropical scarab Oplostomus haroldi (Witte) is a pest of honeybees in East Africa with little information available on its chemical ecology. Recently, we identified a female-produced contact sex pheromone, (Z)-9-pentacosene, from the cuticular lipids that attracted males. Here, we investigated the kairomonal basis of host location in O. haroldi. We used coupled gas chromatography/electroantennographic detection (GC/EAD) and GC/mass spectrometry to identify antennally-active compounds from volatiles collected from honeybee colonies. Antennae of both sexes of the beetle consistently detected seven components, which were identified as 3-hydroxy-2-butanone, 2,3-butanediol, butyl acetate, isopentyl acetate, butyl butyrate, hexyl acetate, and methyl benzoate. In olfactometer bioassays, both sexes responded to the full seven-component synthetic blend over solvent controls, but chose honeybee colony odors over the blend. These findings suggest that the seven compounds are components of a kairomone from honeybee colonies used by O. haroldi.

Chemical communication in the honey bee scarab pest Oplostomus haroldi: role of (Z)-9-pentacosene.

Oplostomus haroldi Witte belongs to a unique genus of afro-tropical scarabs that have associations with honey bee colonies, from which they derive vital nutrients. Although the attributes of the honey bee nest impose barriers to communication among nest invaders, this beetle still is able to detect conspecific mates for reproduction. Here, we show, through behavioral studies, that cuticular lipids serve as mate discrimination cues in this beetle. We observed five steps during mating: arrestment, alignment, mounting, and copulation, and a post-copulatory stage, lasting ~40-70 % of the total mating duration, that suggested mate guarding. Chemical analysis identified the same nine straight-chain alkanes (C(23)-C(31)), six methyl-branched alkanes (6), and five mono-unsaturated alkenes in the cuticular lipids of both sexes. Methyl alkanes constituted the major component (46 %) of male cuticular lipids, while mono-unsaturated alkenes were most abundant (53 %) in females. (Z)-9-Pentacosene was twice as abundant in females than in males, and ~20 fold more concentrated in beetles than in worker bees. In mating assays, (Z)-9-pentacosene elicited arrestment, alignment, and mounting, but not copulation, by male beetles. These results represent the first evidence of a contact sex pheromone in a scarab beetle. Such contact pheromones may be an essential, cryptic mechanism for arthropods associated with eusocial insects.

Estimating reproductive success of Aethina tumida (Coleoptera: Nitidulidae) in honey bee colonies by trapping emigrating larvae.

The small hive beetle (Aethina tumida Murray) is a scavenger and facultative predator in honey bee colonies, where it feeds on pollen, honey, and bee brood. Although a minor problem in its native Africa, it is an invasive pest of honey bees in the United States and Australia. Adult beetles enter bee hives to oviposit and feed. Larval development occurs within the hive, but mature larvae leave the hive to pupate in soil. The numbers leaving, which can be estimated by trapping, measure the reproductive success of adult beetles in the hive over any given period of time. We describe a trap designed to intercept mature larvae as they reach the end of the bottom board on their way to the ground. Trap efficiency was estimated by releasing groups of 100 larvae into empty brood boxes and counting the numbers trapped. Some larvae escaped, but mean efficiency ranged from 87.2 to 94.2%. We envision the trap as a research tool for study of beetle population dynamics, and we used it to track numbers of larvae leaving active hives for pupation in the soil. The traps detected large increases and then decreases in numbers of larvae leaving colonies that weakened and died. They also detected small numbers of larvae leaving strong European and African colonies, even when no larvae were observed in the hives.

Monitoring Aethina tumida (Coleoptera: Nitidulidae) with baited bottom board traps: occurrence and seasonal abundance in honey bee colonies in Kenya.

The population dynamics of the honey bee pest Aethina tumida Murray (small hive beetle) have been studied in the United States with flight and Langstroth hive bottom board traps baited with pollen dough inoculated with a yeast Kodamaea ohmeri associated with the beetle. However, little is known about the population dynamics of the beetle in its native host range. Similarly baited Langstroth hive bottom board traps were used to monitor the occurrence and seasonal abundance of the beetle in honey bee colonies at two beekeeping locations in Kenya. Trap captures indicated that the beetle was present in honey bee colonies in low numbers all year round, but it was most abundant during the rainy season, with over 80% trapped during this period. The survival of larvae was tested in field releases under dry and wet soil conditions, and predators of larvae were identified. The actvity and survival of the beetle were strongly influenced by a combination of abiotic and biotic factors. Larval survival was higher during wet (28%) than dry (1.1%) conditions, with pupation occurring mostly at 0-15 cm and 11-20 cm, respectively, beneath the surface soil during these periods. The ant Pheidole megacephala was identified as a key predator of larvae at this site, and more active during the dry than wet seasons. These observations imply that intensive trapping during the rainy season could reduce the population of beetles infesting hives in subsequent seasons especially in places where the beetle is a serious pest.