Africanized Honey Bees Apis Mellifera Research Articles

Rapid identification of Africanized honey bee Apis mellifera (hymenoptera: apidae) using high-resolution melting analysis DNA from honey

Genetic characterization of honey bee populations, particularly the identification of Africanized honey bees, has traditionally been based on different molecular methods that involve the amplification and analysis of different regions of the mitochondrial or nuclear genome which often take a long time. We introduce high-resolution melting analysis (HRM) as a novel approach for rapid and cost-effective identification of the African lineage in Apis mellifera populations from mitochondrial DNA extracted from honey. Samples were collected from 111 apiaries in 13 Argentinean provinces between 2012 and 2018. Total DNA was extracted from 10 g honey sediment and subjected to quantitative PCR (qPCR) using specific primers (APIS-F/Afr207R) amplifying a 207 bp product. Positive controls (Africanized/Non-Africanized) were used for mitotype validation and a subset of qPCR products were sequenced to compare different haplotypes. HRM analysis enabled the quick detection of sequence polymorphisms in short amplicons generated by qPCR. Among the analyzed apiaries, 26 located above 35° S in Misiones, La Rioja, Catamarca, Formosa, Corrientes, Córdoba, and Santa Fé exhibited the Africanized mitotypes while the remaining apiaries below this latitude, mainly in Buenos Aires, Rio Negro, Neuquén, and Entre Ríos, showed the European mitotypes. These results align with previous characterizations in the region. HRM analysis demonstrated high sensitivity and effectiveness, requiring minimal DNA input, and reducing costs and sampling efforts. This approach provides a rapid and reliable method for identifying the African lineage, facilitating honey bee breeding programs, conservation efforts, and management strategies in regions with Africanized honey bee populations.

Oral administration of phytochemicals protects honey bees against cognitive and motor impairments induced by the insecticide fipronil.

Pollution produced by exposure to pesticides is a major concern for food security because the negative impacts on pollinators. Fipronil, an insecticide broadly used around the globe has been associated with the ongoing decline of bees. With a characteristic neuroactive toxicodynamic, fipronil leads to cognitive and motor impairments at sublethal dosages. Despite of regional bans, multilevel strategies are necessary for the protection of pollinators. Recent evidence suggests that specific nutrients in the diets of bees may induce protection against insecticides. Here, we evaluated whether the administration of three phytochemicals, namely rutin, kaempferol and p-coumaric acid provide protection to the Africanized honey bee Apis mellifera against oral administration of realistic dosages of fipronil. We tested the potential impairment produced by fipronil and the protection induced by the phytochemicals in learning, 24h memory, sucrose sensitivity and motor control. We found that the administration of fipronil induced a concentration-dependent impairment in learning and motor control, but not 24h memory or sucrose sensitivity across a 24h window. We also found that the administration of rutin, p-coumaric acid, kaempferol and the mixture was innocuous and generally offered protection against the impairments induced by fipronil. Overall, our results indicate that bees can be prophylactically protected against insecticides via nutrition, providing an alternative to the ongoing conflict between the use of insecticides and the decline of pollinators. As the studied phytochemicals are broadly present in nectar and pollen, our results suggest that the nutritional composition, and not only its production, should be considered when implementing strategies of conservation via gardens and co-cropping.

A Hitchhiker’s Ride: The Honey Bee Louse Braula Coeca (Diptera: Braulidae) Selects its Host by Eavesdropping

The bee louse Braula spp. had until recently a distribution coincident with its host the honey bee. The adult fly usually attaches to a worker honey bee and steals food from its mouth. However, not all worker bees carry Braula spp. and the mechanism used by Braula spp. to select hosts is not well understood. Using choice remounting bioassays and chemical analyses, we determined host selection and the cues used by B. coeca, a species associated with the African honey bee Apis mellifera scutellata. Braula coeca successfully remounted bees from which they were initially removed and preferred their mandibular gland pheromones (MDG) over those of bees not carrying them. The bee lice did not show any preference for the cuticular hydrocarbons of both types of workers. Chemical analyses of the MDG extracts, revealed quantitative differences between the two categories of workers, with workers carrying B. coeca having more of the queen substance (9-oxo-2(E)-decenoic acid) and worker substance (10-hydroxy-2(E)-decenoic). Braula coeca showed a dose response to the queen substance, indicating its ability to use host derived kairomones as cues that allowed it to benefit from trophallactic dominance by individuals that have a higher probability of being fed by other workers.

Viruses, microsporidia and glyphosate residues in Africanized honey bees Apis mellifera, under field conditions

Bees are affected by several factors, including pathogens and pesticides, which may cause mortality and consequent population decline. Considering the ecological and economic importance of these pollinators, this study evaluated the occurrence of the microsporidia Nosema apis and Nosema ceranae, the viruses ABVP, DWV, BQCV, KBV, IAPV and CBPV and residues of the pesticides atrazine, chlorantraniliprole, deltamethrin, fipronil and glyphosate in workers from field colonies of Africanized honey bee Apis mellifera across one year. The presence of pesticide residues was also evaluated in honey and pollen samples. All tested viruses were found in the workers, with BQCV showing the highest copy number. BQCV levels were correlated with relative humidity. Nosema ceranae was the only microsporidian with higher intensities of infection during summer and autumn; seasons with high availability of food resources, which can act as dispersers of this pathogen. Small amounts of glyphosate and its metabolite were detected in the bees tested. These data show that honey bees are contaminated with pathogens and pesticides in the field and present correlative data for these threats.

Protecting the resource: an assessment of mitigation methods used to protect large trees from African elephant impact in a savanna system

African elephants Loxodonta africana can alter the structural components of savanna ecosystems, often through the reduction of the large tree (≥ 5 m height) cover component. Elephant impact can be amplified in small, protected areas, or areas where water is readily available to elephants. One management option is to protect large trees directly using applied mitigation methods to limit elephant impact. In this paper, we assessed and compared the effectiveness and logistical requirements of four mitigation methods that have been applied to protect large trees from elephant impact in South Africa's Greater Kruger National Park – namely African honeybees Apis mellifera scutellata in beehives; creosote oil in glass jars, concrete pyramids arranged in circles around trees, as well as wire‐netting the trees' main stems. For each method, elephant impact levels and tree mortality rates were measured over a 2–5‐year period depending on the method in use. Sample sizes ranged from 43 to 59 trees per mitigation method, with a comparable control, which was a tree of the same species and morphological dimensions but lacking any mitigation application. Beehives were the most effective method at reducing tree loss, significantly reducing tree mortality from 34% (6.8%/year) in control trees to only 10% (2% year‐1) over the five‐year experimental period. However, beehives were the most expensive method to apply to a tree, although this cost can be compensated through honey sales. Concrete pyramids reduced tree loss when the combined pyramid radius was > 1.5 m in length, whilst wire‐netting was effective against bark‐stripping by elephants but was still vulnerable to heavier forms of impact such as uprooting and stem snapping. Creosote jars did not prevent elephants from impacting treated trees. Our results provide managers with a toolkit for protecting large trees against elephant impact, commenting on both the efficacy and the logistical constraints for each method.

Impact of acute oral exposure to paraquat and glyphosate on food consumption and survival rates of the African honeybee Apis mellifera scutellata Lepeletier (Hymenoptera: Apidae)

Impact of acute oral exposure to paraquat and glyphosate on food consumption and survival rates of the African honeybee Apis mellifera scutellata Lepeletier (Hymenoptera: Apidae)

Diurnality in the defensive behaviour of African honeybees Apis mellifera adansonii and implications for their potential efficacy in beehive fences

AbstractAcross the range of African elephants Loxodonta spp., negative interactions with people are prevalent, and the impact of the resulting economic losses on farmers calls for solutions. The use of beehive fences, a mitigation method with ecological and socio-economic benefits, is gaining momentum in African savannah landscapes. We assessed the diurnal and nocturnal defensive behaviours of African honeybees Apis mellifera adansonii in response to visual and physical disturbances in the Campo–Ma'an conservation area, Cameroon. We examined six bee colonies, assessing their activity level, aggressive behaviour and ability to defend themselves when disturbed at different times of day. We found that activity levels varied between colonies and that colonies were more active during the day and inactive at night. The defensive perimeter around the hives also varied between the colonies and was generally greater during morning and evening periods. Bee colonies did not defend their hives around midday and at night. In response to a threat, bees were more likely to fly out from the hive during daytime than at night, with variation amongst colonies. Overall, as elephant intrusions occur mostly at night, beehive fences alone may not be an adequate mitigation method against crop damage caused by forest elephants Loxodonta cyclotis. We suggest combining beehive fences with other mitigation methods to improve crop protection.

Larval Exposure to Parasitic Varroa destructor Mites Triggers Specific Immune Responses in Different Honey Bee Castes and Species.

Innate immune systems are key defenses of animals and particularly important in species that lack the sophisticated adaptive immune systems as found in vertebrates. Here, we were interested to quantify variation in innate immune responses of insects in hosts that differ in their parasite susceptibility. To do this, we studied immune responses in honey bees, which can host a remarkable number of different parasites, which are major contributors of declining bee health and colony losses. The most significant parasite of honey bees is the mite Varroa destructor, which has infested the majority of global honey bee populations, and its control remains a major challenge for beekeepers. However, a number of nonmanaged honey bees seem able to control Varroa infections, for example, the Eastern honey bee Apis ceranacerana or the African honey bee Apis mellifera scutellata. These bees therefore make interesting study subjects to identify underlaying resistance traits, for example, by comparing them to more susceptible bee genotypes such as Western honey bees (A. melliferaligustica). We conducted a series of interlinked experiments and started with behavioral assays to compare the attractiveness of bee larvae to mites using different honey bee genotypes and castes. We found that 6-day-old larvae are always most attractive to mites, independently of genotype or castes. In a next step, we compared volatile profiles of the most attractive larvae to test whether they could be used by mites for host selection. We found that the abundance of volatile compounds differed between larval ages, but we also found significant differences between genotypes and castes. To further study the expected underlaying physiological differences between potentially resistant and susceptible host larvae, we compared the larval hemolymph proteomes of the three honey bee genotypes and two castes in response to mite exposure. We identified consistent upregulation of immune and stress-related genes in Varroa-exposed larvae, which differed between genotypes and castes. Tolerant honey bee castes and genotypes were characterized by stronger or more distinct immune esponses. In summary, we provide first insights into the complex involvement of the innate immune system of tolerant honey bees against mite infestations, which could be used for future breeding purposes.

Current Status of Acarapis woodi Mite Infestation in Africanized Honey Bee Apis mellifera in Brazil

Acarapis woodi Rennie (Acari: Tarsonemidae) is an endoparasitic mite, which can affect the respiratory system in the honey bee Apis mellifera L. (Hymenoptera: Apidae) causing mortality. This mite was first recorded in Brazil in the 1970s, but its current presence is unclear. Therefore, we evaluated the presence of A. woodi in the Africanized honey bee A. mellifera in Brazil, to update the occurrence data of this parasite, 47 yr after its detection. We examined a total of 153 honey bee colonies, from 15 different states using dissection and molecular techniques. This is the first study of the detection of A. woodi in Brazil using molecular techniques. The results were negative for both methods employed, and we can conclude that A. woodi is not present in Africanized honey bee colonies in Brazil.

The complete mitochondrial genome of the West African honey bee Apis mellifera adansonii (Insecta: Hymenoptera: Apidae)

The complete mitochondrial genome of the West African honey bee Apis mellifera adansonii consisted of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. It was 16,466 bp and consisted of 84.7% AT nucleotides. This subspecies had a similar mitogenome to those of other southern African honey bees, namely A. m. scutellata, A. m. capensis, and A. m. monticola.

Bees biodiversity, forage behavior and fruit production in gherkin crop (Cucumis anguria L.)

The objectives were to evaluate the biodiversity of bees, forage behavior and their effect on fruit production in the gherkin crop (Cucumis anguria L.) in the campus of the University Center Moura Lacerda in two years. The frequency and type of collection of the insects in the flowers was observed by counting from 8:00 a.m. to 5:00 p.m., in the first 10 minutes of each time, for three distinct days in each year. The percentage of fruiting was quantified in 25 female flowers covered with nylon compared to the 25 female flowers uncovered in the two years. The flowers were visited by the Africanized honey bees Apis mellifera and the native bees Plebeia sp., Exomalopsis sp. and Melissodes sp., and the Africanized honey bees presented higher frequency and constancy with a higher number of visits in the male flowers compared to the female ones and these visits occurred between 8:00 a.m. and 3:00 p.m. Without the visitation of the bees there was no fruit production, and both the Africanized honey bee and the native ones when collecting nectar and pollen, visited both female and male flowers, carrying pollen in their body, being considered important pollinators of this culture.

A clinical trial protocol to treat massive Africanized honeybee (Apis mellifera) attack with a new apilic antivenom

BackgroundEnvenomation caused by multiple stings from Africanized honeybees Apis mellifera constitutes a public health problem in the Americas. In 2015, the Brazilian Ministry of Health reported 13,597 accidents (incidence of seven cases per 100,000 inhabitants) with 39 deaths (lethality of 0.25%). The toxins present in the venom, which include melittin and phospholipase A2, cause lesions in diverse organs and systems that may be fatal. As there has been no specific treatment to date, management has been symptomatic and supportive only.MethodsIn order to evaluate the safety and neutralizing capacity of a new apilic antivenom, as well as to confirm its lowest effective dose, a clinical protocol was developed to be applied in a multicenter, non-randomized and open phase I/II clinical trial. Twenty participants with more than five stings, aged more than 18 years, of both sexes, who have not previously received the heterologous serum against bee stings, will be included for 24 months. The proposed dose was based on the antivenom neutralizing capacity and the number of stings. Treatment will be administered only in a hospital environment and the participants will be evaluated for a period up to 30 days after discharge for clinical and laboratory follow-up.ResultsThis protocol, approved by the Brazilian regulatory agencies for ethics (National Commission for Ethics on Research – CONEP) and sanitation (National Health Surveillance Agency – ANVISA), is a guideline constituted by specific, adjuvant, symptomatic and complementary treatments, in addition to basic orientations for conducting a clinical trial involving heterologous sera.ConclusionsThis is the first clinical trial protocol designed specifically to evaluate the preliminary efficacy and safety of a new antivenom against stings from the Africanized honeybee Apis mellifera. The results will support future studies to confirm a new treatment for massive bee attack that has a large impact on public health in the Americas.

Statistical modeling of insect behavioral response to changes in weather conditions in Brassica napus L.

Understanding specific meteorological factors that affect the foraging activity of pollinator insects can provide valuable information to ensure appropriate levels of pollination of crops. This research was carried out to verify the spectrum of pollinating insects and their foraging behavior in relation to specific meteorological variables in the Brassica napus (rapeseed) crop. Data collected consisted of the following observations: number of visits by each pollinator, floral resources collected by Africanized honeybee Apis mellifera L., number of flowers visited by Africanized honeybees in 1 min, number of flowers visited by Africanized honeybees in one specific plant during the timed observation of 1 min, and the time taken by the honeybees to visit each flower. In this study the analyses were made through different generalized linear models. The Africanized honeybees were the most abundant pollinating insects (88%) and most visitors were observed collecting nectar (90%). The Africanized honeybees visited a total of 12.9 ± 1.40 flowers in 1 min and 2.96 ± 1.09 flowers of a single plant in 1 min. The time the honeybees spent on the rapeseed flowers was 4.2 ± 1.6 s. The number of floral visitors correlated closely to the variation of abiotic factors, especially temperature, relative humidity, wind speed, and solar radiation. Africanized honeybees were more active at different times of the day as compared to the other floral visitors and they foraged more intensely on the Hyola 433 hybrid than on the Hyola 61 hybrid. The Hyola 61 hybrid was more attractive to Diabrotica speciosa and Lepidoptera.

Observations of Africanized honey bee Apis mellifera scutellata absence and presence within and outside forests across Rondonia, Brazil

The Africanized honey bee (AHB) has generated much research on invasion ecology and impacts on native pollinators ever since its introduction in Brazil approximately 60 years ago. Some have sought to determine how deforestation plays a role in mediating impacts between the AHB and native pollinators. We present the largest-ever, systematic survey of AHB presence/absence in the Neotropics to test to what extent the AHB prefers to forage within forests vs. outside forests. AHB presence/absence within and outside forest fragments was recorded at 187 locations distributed across the state of Rondonia, Brazil. The landscapes at each location varied in terms of overall level of deforestation and the length of time since modern human settlement. AHB presence was significantly higher outside forest cover. These findings agree with previous studies documenting the spread of AHBs in the Amazon in their preference for deforested areas.

Effects of seasonal variations and collection methods on the mineral composition of propolis from Apis mellifera Linnaeus Beehives.

The effects of seasonal variations and the methods of collection of propolis produced by Africanized honey bees Apis mellifera Linnaeus, 1758, on the composition of constituent minerals such as magnesium (Mg), zinc (Zn), iron (Fe), sodium (Na), calcium (Ca), copper (Cu), and potassium (K) were evaluated. Propolis was harvested from 25 beehives by scraping or by means of propolis collectors (screen, "intelligent" collector propolis [ICP], lateral opening of the super [LOS], and underlay method). During the one-year study, the propolis produced was harvested each month, ground, homogenized, and stored in a freezer at -10 ºC. Seasonal analyses of the mineral composition were carried out by atomic absorption spectrophotometry and the results were evaluated by analysis of variance (ANOVA), followed by Tukey-Kramer's test to compare the mean values (p<0.05). The results showed that seasonal variations influence the contents of 5 minerals (Mg, Fe, Na, Ca, and Cu), and the propolis harvesting method affects the contents of 4 minerals (Mg, Zn, Fe, and Ca).

In vitro effects of thiamethoxam on larvae of Africanized honey bee Apis mellifera (Hymenoptera: Apidae)

Several investigations have revealed the toxic effects that neonicotinoids can have on Apis mellifera, while few studies have evaluated the impact of these insecticides can have on the larval stage of the honeybee. From the lethal concentration (LC50) of thiamethoxam for the larvae of the Africanized honeybee, we evaluated the sublethal effects of this insecticide on morphology of the brain. After determine the LC50 (14.34ng/μL of diet) of thiamethoxam, larvae were exposed to a sublethal concentration of thiamethoxam equivalent to 1.43ng/μL by acute and subchronic exposure. Morphological and immunocytochemistry analysis of the brains of the exposed bees, showed condensed cells and early cell death in the optic lobes. Additional dose-related effects were observed on larval development. Our results show that the sublethal concentrations of thiamethoxam tested are toxic to Africanized honeybees larvae and can modulate the development and consequently could affect the maintenance and survival of the colony. These results represent the first assessment of the effects of thiamethoxam in Africanized honeybee larvae and should contribute to studies on honey bee colony decline.

Hygienic Behavior of Africanized Honey Bees Apis mellifera Directed towards Brood in Old and New Combs during Diurnal and Nocturnal Periods.

Hygienic behavior in honey bees, Apis mellifera, is measured by determining the rate at which the bees uncap and remove dead sealed brood. We analyzed individual behavior of house-cleaning Africanized honey bees in order to focus on some poorly understood aspects of hygienic behavior. Two observation hives, each with approximately 3,000 individually marked bees, were used in this study. The efficiency of hygienic behavior was evaluated in hygienic and non-hygienic strains of bees using two types of combs (new and old), as well as at different periods of the day (night and day). We also recorded the age of workers that performed this task of removing dead brood. In both strains, the workers that performed tasks related to hygienic behavior were within the same age cohort; we found no influence of age on the amount of time dedicated to the task, independent of the type of comb or period of the day. The total time from perforation of the cell capping until the dead brood had been completely removed, and was significantly shorter during daytime than at night. Hygienic behavior directed towards dead brood in new combs was also significantly more efficient (faster) than for brood in old combs. The type of comb had significantly more effect than did the time of day. We conclude that the type of comb and time of day should be taken into consideration when evaluating hygienic behavior in honey bees.

The ectoparasite mite Varroa destructor Anderson and Trueman in southeastern Brazil apiaries: effects of the hygienic behavior of Africanized honey bees on infestation rates

In Brazil, the ectoparasitic mite of bees Varroa destructor Anderson and Trueman (Acari: Varroidae) remains at low levels of infestation causing no major damage. However, with the introduction and possible dominance of a new haplotype (K) of the mite, usually found in areas with high infestation rates (IR), it is necessary to monitor and select beehives that are resistant to the pest in order to avoid future problems. Several factors are listed as potentially being responsible for the dynamics of mite infestation, among which hygienic behavior (HB) stands out. In this context we sought to evaluate the HB of Africanized honey bees Apis mellifera L. (Hymenoptera: Apidae) compared with the mite IR in apiaries of two municipalities of southeastern Brazil (Taubaté and Viçosa). For the municipality of Taubaté, the average IR was 4.9% (3.4 to 5.8%), while the HB averaged 98.6% (96 to 100%). In Viçosa, the average mite IR was found to be 10.0% (5.4 to 21.0%) with an average value for HB of 57.7% (0 to 79.0%). Results from this research show that IR and HB were negatively correlated (R = -0.9627, P&lt;0.01), suggesting that hives with higher HB have lower IR.

Physicochemical and microbiological characterization of cassava flower honey samples produced by africanized honeybees

Cassava producers in the region of Marília-São Paulo are integrating their farming activity with beekeeping to diversify their income. The aim of this study was to evaluate the physicochemical and microbiological quality of honey samples produced by Africanized honeybees Apis mellifera from cassava flower in 2008. Analysis were carried out for pH, total soluble solids (TSS), acidity, moisture, reducing and total sugars, apparent sucrose, hydroxymethylfurfural, color, ash, proteins, water insoluble solids, diastasic activity, mineral content, microbiological evaluations, and mineral and hydrocyanic acid (HCN) content. The honey samples showed physicochemical and microbiological characteristics favorable to commercialization, with the exception of apparent sucrose and acidity, which show the need for a narrow focus of attention to the honey maturation degree at the harvest time and more careful monitoring during production and processing. The commercialization of Brazilian cassava honey, still little explored, can be widely spread in the market since the levels of hydrocyanic acid (HCN) showed no consumption risk; in addition the simultaneous production of honey and cassava provides an alternative to family income increase.

Invasive Africanized honeybees change the structure of native pollination networks in Brazil

The Africanized honeybee Apis mellifera (AHB) is an invasive species spread over all Brazilian biomes, which has negative impacts on native bee populations, but whose impacts on native plants are still controversial. In order to understand how its impacts extend to the pollination service at the community level, we studied the AHB and its interactions in a multi-species context using network theory. We analyzed six pollination networks from the Brazilian Caatinga, a xeric biome where beekeeping is increasing very quickly. The AHB occupied a central position in all networks, as it was responsible for a large share of the interactions observed (14 ± 7 %) and bound together different modules. By simulating the removal of the AHB from each network, we observed no effects on connectance, but a strong decrease in nestedness (−23 ± 19 %) and an increase in modularity (8 ± 5 %). The robustness of networks to cumulative random extinctions was on average not affected. In summary, our evidence points out that the AHB induces significant changes in the structure of native pollination networks, mainly by making them more cohesive and monopolizing many interactions. Although the AHB did not affect network robustness, its net impact on the pollination service may be negative, because this invasive species is very generalistic and may not be an efficient pollinator for some native plants.