Honey Bee Apis Florea Research Articles

Genetic diversity of the Dwarf honeybee (Apis florea Fabricius, 1787) populations based on microsatellite markers.

Apis florea is one of two species of small, wild honeybee. The present study was conducted to evaluate the genetic diversity of Apis florea honeybee from 48 nests (colonies) using microsatellite markers in the South of Iran. All honeybee samples were analyzed for six microsatellite loci (A88, A107, A7, B124, A113 and A35). The six loci had different numbers of alleles in the sampled colonies ranging from 7 (loci A107) to 3 (loci A7, A35). Gene diversity in Apis florea ranged from 0.491 to 0.595. This range probably reflects the spreading of nests in a large region with a varied climate. Phylogenetic tree showed two distinct clusters including a) Minab region samples and b) Bandar Abbas, Bandar Khamir and Qeshm Island regions. All of these regions are geographically rich, having varied vegetation and climate conditions. Our findings are an important contribution to the methods of studying distribution and conservation of Apis florea.

Similar but not the same: insights into the evolutionary history of paralogous sex-determining genes of the dwarf honey bee Apis florea.

Studying the fate of duplicated genes provides informative insight into the evolutionary plasticity of biological pathways to which they belong. In the paralogous sex-determining genes complementary sex determiner (csd) and feminizer (fem) of honey bee species (genus Apis), only heterozygous csd initiates female development. Here, the full-length coding sequences of the genes csd and fem of the phylogenetically basal dwarf honey bee Apis florea are characterized. Compared with other Apis species, remarkable evolutionary changes in the formation and localization of a protein-interacting (coiled-coil) motif and in the amino acids coding for the csd characteristic hypervariable region (HVR) are observed. Furthermore, functionally different csd alleles were isolated as genomic fragments from a random population sample. In the predicted potential specifying domain (PSD), a high ratio of πN/πS=1.6 indicated positive selection, whereas signs of balancing selection, commonly found in other Apis species, are missing. Low nucleotide diversity on synonymous and genome-wide, non-coding sites as well as site frequency analyses indicated a strong impact of genetic drift in A. florea, likely linked to its biology. Along the evolutionary trajectory of ~30 million years of csd evolution, episodic diversifying selection seems to have acted differently among distinct Apis branches. Consistently low amino-acid differences within the PSD among pairs of functional heterozygous csd alleles indicate that the HVR is the most important region for determining allele specificity. We propose that in the early history of the lineage-specific fem duplication giving rise to csd in Apis, A. florea csd stands as a remarkable example for the plasticity of initial sex-determining signals.

Evaluation of anti-inflammatory, anti-nociceptive, and anti-arthritic activities of Indian Apis dorsata bee venom in experimental animals: biochemical, histological, and radiological assessment

Traditionally venoms are used from thousands of years to treat pain, inflammation, and arthritis. In Ayurveda “Suchika Voron” and “Shodhona” were practiced against pain.In the present study, venom composition of the Indian honeybee Apis florea (AF), Apis dorsata (AD), and Apis cerana indica (AC) were analyzed using electrophoresis (SDS-PAGE). This venom analysis was used to shed light upon the correlation in structure and the venom composition among the three species in Indian fields.Among the three species, Indian Apis dorsata bee venom (ADBV) is evaluated for an anti-inflammatory, anti-nociceptive activity, and antiarthritic activity in different animal models. The effect of ADBV is revealed for its anti-arthritic activity in the FCA- and CIA-induced arthritis model in male Wistar rats. The immunosuppressant action of ADBV was studied by hemagglutination antibody titer. It has been found that ADBV possesses anti-inflammatory and antinociceptive activities. In FCA- and CIA-induced arthritis, ADBV able to decrease rheumatoid factor, pain perception parameters, C-reactive protein, erythrocytes sedimentation rate, urinary hydroxyproline, serum transaminase level, and serum nitric oxide level when compared with diseased control arthritic rats. IL-6, TNF-α level was found to be decrease by ADBV treatment in collagen induced arthritis model. Thus this study confirmed the scientific validation behind utilization of venom in Indian Apis dorsata bees in arthritis and inflammatory diseases which has been not reported till date.

Potential Use of an Essential Oil From the Flower of Swertia densifolia as a Repellent for Apis florea (Hymenoptera: Apidae)

Formulations of compounds repellent to honey bees may be useful tools to keep bees away from areas recently treated with toxic insecticides. Here, we report the activity of a previously un- known honey bee repellent, an essential oil from the flower of Swertia densifolia (Gentianaceae), which has activity toward Indian honey bees Apis florea F. The essential oil was obtained by hydrodistillation and was subjected to a rotating table bioassay with foragers of the honey bee A. florea. Repellency was dose-dependent up to 12 mg/ml and remained constant thereafter. The chemical constitution of the essential oil was examined by gas chromatography-mass spectrometry analyses, which identified 19 constituent compounds. The major constituents (>10% of the oil) were linalool and octadecanoic acid. The constituents of intermediate concentration (5-10%) were a-terpeniol, nerol, n-octadecyl acetate, and spirostan-3-ol. Besides these, five minor constituents (1-5%) and six of the eight trace components present (<1%) have also been identified. The response of A. florea foragers to mixtures of the major and intermediate constituents was studied. Screening of formulations of linalool and a-terpeniol showed re- pellent properties toward A. florea, behavior which has already been reported by us in earlier studies. The compound n-octadecyl acetate was found to be an attractant, while octadecanoic acid and spirostan- 3-ol were neither attractants nor repellents. Nerol, however, elicited dose-dependent effects involving both attraction and repellency. The effects of the constituents of this essential oil were not merely additive, possibly due to synergistic effects. These results establish the previously unknown chemical constitution and dose-dependent repellent nature of the essential oil isolated from flower of S. densifolia towardA. florea and may be used to improve bee management.

Bacterial community structure in Apis florea larvae analyzed by denaturing gradient gel electrophoresis and 16S rRNA gene sequencing.

This study characterizes the colonization and composition of bacterial flora in dwarf Asian honeybee (Apis florea) larvae and compares bacterial diversity and distribution among different sampling locations. A. florea larvae were collected from 3 locations in Chiang Mai province, Thailand. Bacterial DNA was extracted from each larva using the phenol-chloroform method. Denaturing gradient gel electrophoresis was performed, and the dominant bands were excised from the gels, cloned, and sequenced for bacterial species identification. The result revealed similarities of bacterial community profiles in each individual colony, but differences between colonies from the same and different locations. A. florea larvae harbor bacteria belonging to 2 phyla (Firmicutes and Proteobacteria), 5 classes (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Bacilli, and Clostridia), 6 genera (Clostridium, Gilliamella, Melissococcus, Lactobacillus, Saccharibacter, and Snodgrassella), and an unknown genus from uncultured bacterial species. The classes with the highest abundance of bacteria were Alphaproteobacteria (34%), Bacilli (25%), Betaproteobacteria (11%), Gammaproteobacteria (10%), and Clostridia (8%), respectively. Similarly, uncultured bacterial species were identified (12%). Environmental bacterial species, such as Saccharibacter floricola, were also found. This is the first study in which sequences closely related to Melissococcus plutonius, the causal pathogen responsible for European foulbrood, have been identified in Thai A. florea larvae.

A study on biochemical composition of the sting gland (poison gland) and the reservoir (poison sac) of the dwarf honey bee Apis florea F. workers

The glands associated with the sting apparatus of worker honey bee Apis florea produce Venom which is known to be composed of a wide spectrum of biomolecules ranging from biogenic amines to peptides and proteins. The Venom apparatus showed the presence of two important associated exocrine glands i.e. Venom gland and Dufors gland. The secretions of both glands are apocrine and are released into the lumen to be stored in the venom sac. The presence of some exocrine cells in the distal part of venom sac which is otherwise known to only store the component of Venom gland led to the present study. The present study that there were considerable differences in the biochemical composition of Venom gland and Venom sac secretions of Apis species The concentration of lipids (Sting gland =1.423±0.0001 and Reservoir = 1.21±0.0067), proteins (Sting gland=0.440±0.0226, Reservoir = 0.390± 0.032), activity of acid phosphatase (Sting gland=112.09±21.100, Reservoir=22.63±1.467) and hexokinase (Sting gland=20.7±4.016, Reservoir=10.66±2.465) was found to be more in case of Venom gland while cholesterol(Sting gland=0.138±0.0161 reservoir=0.324±0.00323), glucose (Sting gland=189±1.31, Reservoir=321±7.19), free amino acids, and activity of alkaline phosphatase (Sting gland=21.03±0.195 Reservoir=22.4±0.685) was more in Venom sac. Glycogen was absent in both Venom gland and Venom sac of Apis species as confirmed by the absence of glucose-6-phosphatase activity. It is established from the present study that Venom sac also secretes various biochemicals and enzymes which are added to the total Venom.

Morphometric Studies on Dwarf Honey Bee Apis Florea F. Workers in Saudi Arabia

Abstract Here we performed a morphometric analysis of dwarf honey bee (Apis florea F.) workers from Saudi Arabia, and we detected morphometric variations within the species in the Jubail and Al-Ahsa provinces of this region. Between the two provinces, Jubail is further north and is closer to sea level, and data showed that during the collection period, Jubail experienced significantly higher humidity. Compared to workers from the Jubail province, workers from Al-Ahsa province showed significantly higher body weight, measured head appendage characteristics, wing characteristics (apart from hindwing width), leg characteristics, and length and width of the 3rd abdominal sternite and 1st wax mirror. Body weight as found to be significantly positively correlated with all determined characteristics, except for flagellum length, hindwing width, 3rd tergite length, and lengths of the 4th tergite and sternite. It can be concluded that body weight and morphometric characteristics of dwarf honey bee workers were significantly affected by geographic origin. The morphometric measurements were within the range reported from most previously examined countries.

Evaluation of catecholamines and amino acids from venom reservoir extract of Indian honey bee (Apis) species

SummaryCatecholamine systems play a variety of roles in animals as neurotransmitters, neuromodulators, hormones, paracrine and autocrine factors. Many venoms of widely different animals have evolved to manipulate these systems for both defensive and offensive purposes. Catecholamines are sympathomimetic and comprise of a catechol group derived from the amino acid tyrosine. Physiological and anatomical evidence suggests that certain amino acids act as neurotransmitters of the major excitatory or inhibitory commands in the central nervous system, and also take part in intermediary metabolism. Catecholamines were quantified from an extract of the venom gland and reservoir of three honey bee species, Apis cerana, Apis dorsata and Apis florea using liquid chromatography with electrochemical detection (ECD). The results showed species dependent variation: 1. norepinephrine and dopamine were found in all the three Apis species, with the highest quantity in A. dorsata followed by A. cerana and the least in A. florea; 2. 5-hydroxyindole-3-acetic-acid was found only in A. cerana; and 3. 5 -hydroxytryptamine was present only in A. dorsata and A. florea. The other two vital bioactive components examined, 3,4 dihydroxyphenyl acetic acid and hydroxy-3-methoxy-phenylacetic acid were absent from all three bee species. Eight amino acids (aspartate, glutamate, serine, alanine, glysine, phenyl ethanolamine, gamma-aminobutyric acid and tyrosine) were analysed and compared, showing that the quantity of each amino acid in the venoms differed between bee species.

Relative Toxicity of Fenazaquin Against Two-Spotted Spider Mite on Okra

Continuous use of previous generation acaricides in okra [Abelmoschus esculentus (L.) Moench] has increased resistance to pesticides and pest resurgence. The project was undertaken to evaluate a new acaricide, fenazaquin 10 EC, for effective control of two-spotted spider mites [Tetranychus urticae Koch (Acari: Tetranychidae)] in pot culture and under field conditions. Fenazaquin was used at 75, 100, 125, or 150 g·ha−1 a.i. and compared with the standard acaricide dicofol 18.5 EC at 250 g·ha−1 a.i. and an untreated control. The adulticide bioassay, ovicidal action to T. urticae and safety of Dwarf honey bees (Apis florea Fabricius), was performed under laboratory conditions. Pot culture and two field trials were conducted to evaluate efficacy. Impact on okra yield and phytotoxicity were determined. The LC50 was 1.440 mg·L−1 and LT50 at 1.50 mg·L−1 was ˜8 hours. Fenazaquin produced more than 70% egg mortality in all doses for T. urticae. Fenazaquin at 125 and 150 g·ha−1 a.i. caused the highest reduction in numbers of mites in pot culture and field experiments. Fruit yields (9.89 Mt·ha−1, 10.59 Mt·ha−1) and cost–benefit ratio (1:3.25, 1:3.48) were higher in fenazaquin-treated plants at 150 g·ha−1 a.i in field experiments. No test doses were phytotoxic and all doses were relatively safe to Dwarf honey bees.

Essential Oil of Indian Propolis: Chemical Composition and Repellency against the Honeybee Apis florea

Hitherto unknown biological properties and the chemical composition of the essential oil isolated from propolis of Indian origin were established. GC/MS Analysis of the essential oil revealed the presence of 32 constituents, of which ten were major compounds, nine had intermediate contents, and 13 were minor compounds. With the exception of six minor constituents, that could not be identified, their identification was based on the comparison of their mass spectra and Kovats retention indices with those listed in the NIST and Wiley mass spectral libraries. Their structural assignment was confirmed by GC/MS co-injection of the essential oil with authentic compounds. Quantification of the components was done by GC-FID analyses. Moreover, the essential oil was shown to possess repellent activity against the honeybee Apis florea. The activity was found to be dose dependent. The average repellency (ΔR) increased with increasing essential-oil concentration up to 24 μg/ml and remained constant for the formulation with the higher concentration. These findings established the chemical constitution of the essential oil and might be useful to beekeepers for the improvement of the bee management.

Susceptibility of four different honey bee species to Nosema ceranae

In this study, we investigated the infectivity of Nosema ceranae and the immune response of the European honey bee, Apis mellifera and the Asian honey bee species, Apis cerana, Apis dorsata and Apis florea when inoculated with two isolates of N. ceranae isolated from different climates (Canada and Thailand), using cage experiments. The results indicated that the local isolate of N. ceranae (Thailand) had high infectivity in A. mellifera, A. cerana and A. dorsata but only a few spores were observed in A. florea. However, we found that only two honey bee species, A. mellifera and A. dorsata became infected when inoculated with N. ceranae isolated from Canada. Finally, our results showed that transcript levels of antimicrobial peptides (AMPs) in Asian honey bees were significantly higher than that of A. mellifera in both the control and N. ceranae inoculated bee groups. Comparing the expression of AMPs between the control and inoculated bees in each species, it was evident that N. ceranae inoculations did not affect the expression level of abaecin in all four honey bees species investigated in this experiment. Nevertheless, we found a significant up-regulation of apidaecin in A. cerana and A. florea when inoculated with N. ceranae (Canadian isolate). Also, the mRNA levels of hymenoptaecin were significantly increased in A. cerana after inoculation by N. ceranae isolated from Canada as compared with the Thai isolate.

Associative olfactory learning of the red dwarf honey bee Apis florea

Apis florea has a wide distribution in southeast Asia, where it is the dominant honey bee species. It is an excellent colonizer, suggesting advanced behavioral plasticity. Using the proboscis extension response (PER) conditioning method, we investigated A. florea associative olfactory learning ability and compared it to the learning performances of Apis mellifera. A. florea workers reacted to pure odorant (conditioned stimulus) only when it was delivered in paired associations with sucrose rewards (unconditioned stimulus), and discriminated between a rewarded and unrewarded odor. Moreover, A. florea levels of acquisition in olfactory associative learning were higher in our study than those of A. mellifera, and they showed better retention of learning after 24 h than A. mellifera. Our results show that A. florea can be studied using PER procedures, enlarging the scope for future comparative studies of associative olfactory learning ability in bees.

First evidence of hygienic behaviour in the dwarf honey bee Apis florea

The hygienic behaviour of eight A. florea honey bee colonies was investigated in Thailand. Sealed brood was deep frozen or killed using entomological pins. The pins were 0.2 or 0.7 mm in diameter. The removal of killed brood by the bees was checked in intervals ranged from 24 to 60 h after it had been presented to the colonies. Almost eighty nine (89%) of the freeze-killed brood was removed by the bees within 24 h after being introduced into the combs, while 95% of the pin-killed brood was removed within 48 h after introduction. Thus, the A. florae colonies could be recognized as hygienic honey bees.

Phylogenetic analysis of Nosema ceranae isolated from European and Asian honeybees in Northern Thailand

Nosema ceranae was found to infect four different host species including the European honeybee ( A. mellifera) and the Asian honeybees ( Apis florea, A. cerana and Apis dorsata) collected from apiaries and forests in Northern Thailand. Significant sequence variation in the polar tube protein (PTP1) gene of N. ceranae was observed with N. ceranae isolates from A. mellifera and A. cerana, they clustered into the same phylogenetic lineage. N. ceranae isolates from A. dorsata and A. florea were grouped into two other distinct clades. This study provides the first elucidation of a genetic relationship among N. ceranae strains isolated from different host species and demonstrates that the N. ceranae PTP gene was shown to be a suitable and reliable marker in revealing genetic relationships within species.

Deciding on the wing: in-flight decision making and search space sampling in the red dwarf honeybee Apis florea

During reproductive swarming and seasonal migration, a honeybee swarm needs to locate and move to a new, suitable nest site. While the nest-site selection process in cavity-nesting species such as the European honeybee Apis mellifera is very precise with the swarm carefully selecting a single site, open-nesting species, such as Apis florea, lack such precision. These differences in precision in the nest-site selection process are thought to arise from the differing nest-site requirements of open- and cavity-nesting species. While A. florea can nest on almost any tree, A. mellifera is constrained by the scarcity of suitable nest sites. Here we show that imprecision in the nest-site selection process allows swarms to quickly reach a decision when many nest sites are available. In contrast, a very precise nest-site selection process slows down the decision-making process when nest sites are abundant.

Bioassay of the mandibular gland pheromones of Apis florea on the foraging activity of dwarf honey bees

SummaryIn this study, we tested the responses (attractiveness or repellence) of two species of dwarf honey bee (Apis andreniformis and Apis florea) foragers to the synthetic mandibular gland pheromones 1-eicosanol, eicosane, heneicosane and 2-heptanol. The pheromone concentrations were varied from 0.0, 0.1, 0.5, 1.0, 5.0 and 10.0% (v/v) in n-hexane. A. andreniformis foragers were repelled by 5.0% eicosane and 5.0 and 10.0% heptanol, while 0.5 and 1.0% 1-eicosanol, 0.1, 0.5, 1.0, 5.0 and 10.0% eicosane, 0.5 and 1.0% heneicosane and 0.1, 0.5, 1.0, 5.0 and 10.0% 2-heptanol repelled A. florea foragers. A. andreniformis foragers were attracted to 0.1% 1-eicosanol, 10.0% eicosane, 1.0, 5.0 and 10.0% heneicosane and 0.1% 2-heptanol. None of the pheromones were attractive to A. florea foragers.

Moving home: nest-site selection in the Red Dwarf honeybee (Apis florea)

The Red Dwarf honeybee (Apis florea) is one of two basal species in the genus Apis. A. florea differs from the well-studied Western Hive bee (Apis mellifera) in that it nests in the open rather than in cavities. This fundamental difference in nesting biology is likely to have implications for nest-site selection, the process by which a reproductive swarm selects a new site to live in. In A. mellifera, workers show a series of characteristic behaviors that allow the swarm to select the best nest site possible. Here, we describe the behavior of individual A. florea workers during the process of nest-site selection and show that it differs from that seen in A. mellifera. We analyzed a total of 1,459 waggle dances performed by 197 scouts in five separate swarms. Our results suggest that two fundamental aspects of the behavior of A. mellifera scouts—the process of dance decay and the process of repeated nest site evaluation—do not occur in A. florea. We also found that the piping signal used by A. mellifera scouts to signal that a quorum has been reached at the chosen site, is performed by both dancing and non-dancing bees in A. florea. Thus, the piping signal appears to serve a different purpose in A. florea. Our results illustrate how differences in nesting biology affect the behavior of individual bees during the nest-site selection process.

Infections of Nosema ceranae in four different honeybee species

The microsporidium Nosema ceranae is detected in honeybees in Thailand for the first time. This endoparasite has recently been reported to infect most Apis mellifera honeybee colonies in Europe, the US, and parts of Asia, and is suspected to have displaced the endemic endoparasite species, Nosema apis, from the western A. mellifera. We collected and identified species of microsporidia from the European honeybee ( A. mellifera), the cavity nesting Asian honeybee ( Apis cerana), the dwarf Asian honeybee ( Apis florea) and the giant Asian honeybee ( Apis dorsata) from colonies in Northern Thailand. We used multiplex PCR technique with two pairs of primers to differentiate N. ceranae from N. apis. From 80 A. mellifera samples, 62 (77.5%) were positively identified for the presence of the N. ceranae. Amongst 46 feral colonies of Asian honeybees ( A. cerana, A. florea and A. dorsata) examined for Nosema infections, only N. ceranae could be detected. No N. apis was found in our samples. N. ceranae is found to be the only microsporidium infesting honeybees in Thailand. Moreover, we found the frequencies of N. ceranae infection in native bees to be less than that of A. mellifera.

Essential Oil of Terminalia chebula Fruits as a Repellent for the Indian Honeybee Apis florea

Plant-based repellent formulations for honeybees play an important role in the bee management. For this purpose, the essential oil of an Indian medicinal plant, Terminalia chebula, commonly known as Myrobalan, was isolated for the first time. Hitherto unknown chemical constituents of the essential oil were determined by GC/MS. The repellent activity of formulations of the essential oil, tested towards the Indian honeybee Apis florea, was found to be dose dependent. The repellency (DeltaR) increased with the concentration of essential oil in the formulations, reaching a maximum for the formulation containing 50 mg/ml of oil. A further increase in the oil concentration was found to reduce the DeltaR. The screening of formulations of the major essential-oil components identified indicated that formulations of furfural, 5-methylfurfural, tetradecanoic acid, palmitic acid, and oleic acid elicited no response in honeybees. In contrast, the formulations of phenylacetaldehyde were repellent, while those of ethyl cinnamate were attractant. These findings might be an asset for beekeepers to improve the bee management. Attractant formulations are effective to attract bees to the desired areas, thus improving the efficiency of pollination. Repellent formulations are used to repel honeybees, especially when toxic insecticides are sprayed on the fields.

Lack of interspecific parasitism between the dwarf honeybees Apis andreniformis and Apis florea

The dwarf honeybees Apis florea and Apis andreniformis are sympatric in Southeast Asia. We examined undisturbed nests of both species finding that heterospecific workers are present in some nests at low frequency. This suggested that workers may enter heterospecific nests as a prelude to reproductive parasitism. To test this hypothesis, we created mixed-species colonies and determined the reproductive response of workers within them based on molecular markers. In queenless colonies, workers of both species activated their ovaries at equal frequency. However, the majority species, A. florea, had complete reproductive dominance over A. andreniformis, most likely because the A. florea workers recognised and removed heterospecific larvae. In queenright mixed-species colonies, workers responded to heterospecific signals of the presence of the queen and did not activate their ovaries. Thus, despite predictions from kin selection theory that workers would benefit from parasitising heterospecific nests, we find no evidence that selection has established a parasitic strategy in these sibling species.