Brood Chamber Research Articles

An Evaluation of the Honey Bee (Hymenoptera: Apidae) Safety Profile of a New Systemic Insecticide, Flupyradifurone, Under Field Conditions in Florida.

Flupyradifurone (Sivanto) is a novel systemic insecticide from the butenolide class developed by Bayer. Based on available data (USEPA 2014), this insecticide appears to have a favorable safety profile for honey bee colonies. As a result, the label permits the product to be applied during prebloom and bloom in various crops, including citrus, except when mixed with azole fungicides during the blooming period. We placed 24 honey bee (Apis mellifera L.) colonies adjacent to eight flowering buckwheat (Fagopyrum esculentum Moench) fields that either had been sprayed with the maximum label rate of flupyradifurone or with water only (control fields), with three colonies placed adjacent to each field. We conducted colony strength assessments during which the number of adult bees, eggs, uncapped brood cells, capped brood cells, food storage cells, and weights of honey supers and brood chambers were determined prior to, during, and after the flowering period. We also analyzed bee-collected pollen and nectar for flupyradifurone residues. Overall, there were no differences in any colony strength parameter for colonies placed at control and flupyradifurone-treated buckwheat fields. Residue analyses showed that pollen (x = 565.8 ppb) and nectar (x = 259.4 ppb) gathered by bees on fields treated with flupyradifurone contained significantly higher flupyradifurone residues than did bee bread and unprocessed nectar collected by bees from control fields (75% of samples <LOD). Within the conditions set forth by our experimental design, our collective data suggest no adverse effects of flupyradifurone on honey bee colonies when following label directions.

Nest Architecture of Tetragona clavipes (Fabricius) (Hymenoptera, Apidae, Meliponini)

The aim of this study was to describe the nest architecture of Tetragona clavipes colonies, and to compare its features with other stingless bee species. The measurements were made on 10 colonies in terms of brood chamber, brood cells, royal cells, food pots, pillars between combs, and population. Brood chamber (= 3117.9 ± 1230.6 cm3) of spiral-shaped combs were all in the center of the nest. The values of equatorial diameter of brood chambers (= 14.33 ± 3.03 cm) can be useful to scale bee boxes. The height and width of food pots (= 3.42 ± 0.39 cm; = 2.21 ± 0.23 cm), and brood cells (= 6.37 ± 0.32 mm; = 3.73 ± 0.35 mm) were statistically different among colonies. The measurements of the internal nest structures of T. clavipes were similar to the few other reported studies of this species. T. clavipes nests presented only few differences from other stingless bee species. This species has large potential for honey production, which has not yet been explored and deserves attention in future studies.

Muscle system in rediae and daughter sporocysts of several digeneans

Morphological studies on digenetic trematodes are quite numerous, but the large majority of researchers deal with the hermaphroditic generation (cercariae, metacercariae, adults). Parthenitae (miracidia, sporocysts, rediae), although they constitute a significant part of a digeneans’ life cycle, attract much less attention. The sparse literature concerning the musculature of parthenitae indicates that it differs in many respects from that of digenean hermaphroditic generation and many other flatworms. We have tried to fill the knowledge gap on digenean muscle systems by focusing on daughter parthenitae (three species with rediae and two with daughter sporocysts). The study was performed using TRITC-phalloidin labeling and confocal microscopy and was aimed at both describing general trends and comparing different morphotypes. The basic body-wall muscle layers were confirmed to be composed of circular and longitudinal muscle fibers. Circular fibers form quite a uniform layer, and longitudinal fibers are typically joined into bundles. The density of muscle layers varies widely among studied species, and the possible causes of this variation are discussed. The internal muscle fibers and bundles are present in all studied species, being most prominent in the anterior region. The brood chamber and birth pore channel often have a muscular lining. The rediae pharynx has a typical set of muscle elements, mostly coinciding with one in the pharynx of the hermaphroditic generation. Taken together, our findings suggest that data on the musculature of daughter parthenitae are important from an evolutionary perspective.

Effects of multiple climate change stressors: ocean acidification interacts with warming, hyposalinity, and low food supply on the larvae of the brooding flat oyster Ostrea angasi

Ocean acidification, rising temperatures, and increased intensity of rain events are occurring due to climate change. Individually, each of these stressors has the potential to influence the growth and survival of many marine organisms, particularly during early development. Together the interactive and multiple impacts of elevated pCO2, temperature, and salinity may be exacerbated by a lack of food. Life history traits are important in determining the response of organisms to climate change. Larvae that develop within a brood chamber, such as the flat oyster, Ostrea angasi, may be pre-exposed to living a higher CO2 environment. This study determined the pH of the fluid surrounding the gills of adult oysters where larvae are brooded and investigated the interactive effects of the multiple climate-related stressors: ocean acidification, warming, hyposalinity, and reduced food availability, on development of O. angasi larvae. The fluid surrounding the larvae was of pH 7.88 ± 0.04, lower than that of surrounding sea water, and was significantly reduced (to pH 7.46 ± 0.05) when oysters remained closed as occurs in nature during periods of stress caused by low salinity. Elevated pCO2 [853–1194 µatm (pHNBS 7.79)] resulted in larvae being 3 % smaller, but it had no effect on the timing of progression through developmental stages, percentage of abnormalities, or survival of larvae. Exposure to elevated pCO2 together with increased temperature (+4 °C) or reduced salinity (20) had a negative effect on the time to the eyed larval stage and with an increase in the percentage of abnormal larvae. Unexpectedly, larvae did not meet their higher metabolic requirements to survive under elevated pCO2 by eating more. In a sublethal effect of elevated pCO2, larval feeding was impaired. We found that O. angasi larva were relatively resilient to elevated pCO2, a trait that may be due to the acclimatisation of hypercapnic conditions in the brood cavity or because they are released from the brood cavity at an older, possibly less sensitive stage. This result contrasts with the larvae of broadcast spawning oysters which are extremely sensitive to elevated pCO2.

Fatty Acids of Densely Packed Embryos ofCarcinus maenasReveal Homogeneous Maternal Provisioning and No Within-Brood Variation at Hatching

Embryonic development of decapod crustaceans relies on yolk reserves supplied to offspring through maternal provisioning. Unequal partitioning of nutritional reserves during oogenesis, as well as fluctuating environmental conditions during incubation, can be sources of within-brood variability. Ultimately, this potential variability may promote the occurrence of newly hatched larvae with differing yolk reserves and an unequal ability to endure starvation and/or suboptimal feeding during their early pelagic life. The present study evaluated maternal provisioning by analyzing fatty acid (FA) profiles in newly extruded embryos of Carcinus maenas Also assessed were the dynamics of such provisioning during embryogenesis, such as embryo location within the regions of the brooding chamber (left external, left internal, right external, and right internal). The FA profiles surveyed revealed a uniform transfer of maternal reserves from the female to the entire mass of embryos, and homogeneous embryonic development within the brooding chamber. Although C. maenas produces a densely packed mass of embryos that are unevenly distributed within its brooding chamber, this factor is not a source of within-brood variability during incubation. This finding contrasts with data already recorded for larger-sized brachyuran crabs, and suggests that the maternal behavior of C. maenas promotes homogeneous lipid catabolism during embryogenesis.

Reproductive Biology and Quantity Evaluation of the Black-Striped Pipefish &lt;i&gt;Syngnathus abaster&lt;/i&gt; (Eichwald, 1831) in the Zaporozhian Reservoir

The article presents the results of biology and ecology studies about the black-striped pipefish Syngnathus abaster (Eichwald, 1831) in the Zaporozhian Reservoir. Quantity and biomass of fish fingerlings and yearlings in different parts of the reservoir was determined. For the first time the reproductive indicators of pipefish in Zaporozhian Reservoir were investigated: the timing of spawning, fish fertility analysis, information on the number of embryos in the brood chamber of males were shown.

Reproductive Biology and Quantity Evaluation of the Black-Striped Pipefish &lt;i&gt;Syngnathus abaster&lt;/i&gt; (Eichwald, 1831) in the Zaporozhian Reservoir

The article presents the results of biology and ecology studies about the black-striped pipefish Syngnathus abaster (Eichwald, 1831) in the Zaporozhian Reservoir. Quantity and biomass of fish fingerlings and yearlings in different parts of the reservoir was determined. For the first time the reproductive indicators of pipefish in Zaporozhian Reservoir were investigated: the timing of spawning, fish fertility analysis, information on the number of embryos in the brood chamber of males were shown.

Warfare in stingless bees

Bees are well known for being industrious pollinators. Some species, however, have taken to invading the nests of other colonies to steal food, nest material or the nest site itself. Despite the potential mortality costs due to fighting with an aggressive opponent, the prospects of a large bounty can be worth the risk. In this review, we aim to bring together current knowledge on intercolony fighting with a view to better understand the evolution of warfare in bees and identify avenues for future research. A review of literature reveals that at least 60 species of stingless bees are involved in heterospecific conflicts, either as attacking or victim colonies. The threat of invasion has led to the evolution of architectural, behavioural and morphological adaptations, such as narrow entrance tunnels, mud balls to block the entrance, decoy nests that direct invaders away from the brood chamber, fighting swarms, and soldiers that are skilled at immobilising attackers. Little is known about how victim colonies are selected, but a phylogenetically controlled analysis suggests that the notorious robber bee Lestrimelitta preferentially attacks colonies of species with more concentrated honey. Warfare among bees poses many interesting questions, including why species differ so greatly in their response to attacks and how these alternative strategies of obtaining food or new nest sites have evolved.

Identification and Expression Analysis of Upregulated Genes in the Resting Egg-Producing Water Flea (Daphnia pulex).

Water fleas (Daphnia pulex) normally produce subitaneous eggs that initiate development immediately after oviposition. However, in response to habitat degradation, resting eggs are produced, which are enclosed in a sturdy outer envelope (ephippium) and can survive in harsh environments for an extended time. To understand the molecular mechanism underlying resting egg production in D. pulex, we investigated the genes whose expression patterns played a role in the production and identified the following six candidate genes: Dpfa-1, Dpfa-2, Dpep-1, Dpep-2, Dpep-3, and Dpep-4. These six genes displayed > 40-fold higher expression levels in resting egg-producing animals compared with those in subitaneous egg-producing animals at the period when the ovaries were mature. Dpfa-1 and Dpfa-2 were expressed in the fat cells, and their expression patterns were synchronized with the development of resting egg oocytes in the ovary. In contrast, Dpep-1-4 were expressed in the morphologically altered epidermal cells of the brood chamber with the formation of the ephippium, and their expression patterns were also related to ephippium formation. Our results suggest that the former two genes encode the resting egg-specific components produced by fat cells and that the latter four genes encode the components related to the ephippium formation synthesized by epidermal cells.

Do well-integrated species of an inquiline community have a lower brood predation tendency? A test using red wood ant myrmecophiles

BackgroundA host infected with multiple parasitic species provides a unique system to test evolutionary and ecological hypotheses. Different parasitic species associated with a single host are expected to occupy different niches. This niche specialization can evolve from intraguild competition among parasites. However, niche specialization can also be structured directly by the host when its defence strategy depends on the parasite’s potential impact. Then it can be expected that species with low or no tendency to prey on host brood will elicit less aggression than severe brood parasitic species and will be able to integrate better in the host system.We examined this hypothesis in a large community of symbionts associated with European red wood ants (Formica rufa group) by testing the association between 1) level of symbiont integration (i.e. presence in dense brood chambers vs. less populated chambers without brood) 2) level of ant aggression towards the symbiont 3) brood predation tendency of the symbiont.ResultsSymbionts differed vastly in integration level and we demonstrated for the first time that relatively unspecialized ant symbionts or myrmecophiles occur preferentially in brood chambers. Based on their integration level, we categorize the tested myrmecophiles into three categories: 1) species attracted to the dense brood chambers 2) species rarely or never present in the brood chambers 3) species randomly distributed throughout the nest. The associates varied greatly in brood predation tendency and in aggression elicited. However, we did not find a correlation for the whole myrmecophile community between a) brood predation tendency and host’s aggression b) integration level and host’s aggression c) integration level and brood predation tendency.ConclusionsOur results indicate that red wood ants did not act more hostile towards species that have a high tendency to prey on brood compared to species that are less likely or do not prey on brood. We show that potentially harmful parasites can penetrate into the deepest parts of a social insect fortress. We discuss these seemingly paradoxical findings in relation to models on coevolution and evolutionary arms races and list factors which can make the presence of potentially harmful parasites within the brood chambers evolutionary stable.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-016-0583-6) contains supplementary material, which is available to authorized users.

Influence of the presence of Cacoxenus indagator Loew. parasite larvae in brood chambers on the emergence rate and size of red mason bees

Cacoxenus indagator is one of cleptoparasites most frequently found in the nests of Osmia bicornis L. The goal of this experiment was to examine the influence of the presence of 2-3 C. indagator larvae in the brood chamber on the cocoon mass, on the mass and size of bee imagines, and on their emergence rate. During the analysis of red mason bee nest material, 200 cocoons were taken from brood chambers, each of them also containing 2-3 larvae of C. indagator (CC). The control group consisted of 200 randomly chosen cocoons from brood chambers with no parasites inside (CFFC). The cocoons and the emerged bees were weighed, and then the size of the bees was determined by the microscope image analysis software Axio Vision Rel. 4.0 coupled with a Stereo Lumar V12 stereoscopic microscope (Carl Zeiss, Germany). This involved measuring the sum of the widths of tergites 3 and 4, the distance between the wings and the forewing length and width. It was found that the presence of 2-3 C. indagator larvae in the brood chamber had no impact on the mortality of bees in cocoons. The research demonstrates that CC cocoons do not have to be removed when collecting cocoons from artificial nests in managed O. bicornis populations, as bees emerging from such cocoons are fully developed.

Fossilized-Brood Chambers in Nummulites vascus from Libya

Spherical calcite chamber-like objects of almost identical size have been found in a few tests of Nummulites vascus. These Nummulites constitute the dominant benthic foraminifera assembly among the reported foraminiferal assemblage of the Abraq Formation (Late Oligocene) at the Daryanah- Al Abyar Roadcut in Northeast Libya. These structures are identified as brood chambers or cysts formed during reproduction and retained within the mother test, where the embryos were hosted during life. The possibility of a diagenetic origin to these cysts is excluded due to the hollow-nature of these objects which supports a biotic origin rather than diagenesis. Similarly, a symbiotic relationship with microalga is also excluded due to the lack of any cellular microstructure. The existence of calcitic brood chambers in Nummulites is hitherto unknown. Importantly this sample appears to be the first example of a calcified brood pouch fossil or ‘cyst’ in Nummulites (23– 28 Ma) research.

Residues of neonicotinoids and their metabolites in honey and pollen from sunflower and maize seed dressing crops

A study was carried out to evaluate the possible presence of thiamethoxam, clothianidin and imidacloprid, as well as the metabolic breakdown products of these three neonicotinoids in pollen and honey obtained from brood chamber combs of honeybee colonies located next to sunflower and maize crops from coated seeds. Samples were analyzed by liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry detector, in combination with accurate mass tools such as diagnostic ions by exact mass, chlorine mass filters, and MS/MS experiments. The presence of thiamethoxam and clothianidin was confirmed in some of the pollen samples analyzed. Moreover, different metabolites of neonicotinoids were tentatively detected in the pollen and honey samples collected. The results suggested that four metabolites were found in the honey samples, while for pollen samples eleven metabolites were identified; among these, five were considered for the first time as metabolic breakdown products in sunflower and maize plants.

The Unique Dorsal Brood Pouch of Thermosbaenacea (Crustacea, Malacostraca) and Description of an Advanced Developmental Stage of Tulumella unidens from the Yucatan Peninsula (Mexico), with a Discussion of Mouth Part Homologies to Other Malacostraca.

The Thermosbaenacea, a small taxon of crustaceans inhabiting subterranean waters, are unique among malacostracans as they brood their offspring dorsally under the carapace. This habit is of evolutionary interest but the last detailed report on thermosbaenacean development is more than 40 years old. Here we provide new observations on an ovigerous female of Tulumella unidens with advanced developmental stages in its brood chamber collected from an anchialine cave at the Yucatan Peninsula, which is only the third report on developmental stages of Thermosbaenacea and the first for the genus Tulumella. Significant in a wider crustacean context, we report and discuss hitherto unexplored lobate structures inside the brood chamber of the female originating at the first (maxilliped) and second thoracic segments, which are most likely modified epipods, perhaps serving as gills. At the posterior margin of carapace of the female are rows of large spines preventing the developing stages from falling out. The external morphology of the advanced developmental stages is described in much detail, providing information on e.g., carapace formation and early limb morphology. Among the hitherto unknown structures in the advanced developmental stages provided by this study are the presence of an embryonic dorsal organ and rudimentary ‘naupliar processes’ of the second antennae. Since most hypotheses on crustacean (and malacostracan and peracaridan) relationship rest on external limb morphology, we use early limb bud morphology of Tulumella to better establish thermosbaenacean limb homologies to those of other crustaceans, which is a necessary basis for future morphology based phylogenetic considerations.

Aggressive mimicry coexists with mutualism in an aphid

Understanding the evolutionary transition from interspecific exploitation to cooperation is a major challenge in evolutionary biology. Ant-aphid relationships represent an ideal system to this end because they encompass a coevolutionary continuum of interactions ranging from mutualism to antagonism. In this study, we report an unprecedented interaction along this continuum: aggressive mimicry in aphids. We show that two morphs clonally produced by the aphid Paracletus cimiciformis during its root-dwelling phase establish relationships with ants at opposite sides of the mutualism-antagonism continuum. Although one of these morphs exhibits the conventional trophobiotic (mutualistic) relationship with ants of the genus Tetramorium, aphids of the alternative morph are transported by the ants to their brood chamber and cared for as if they were true ant larvae. Gas chromatography-mass spectrometry analyses reveal that the innate cuticular hydrocarbon profile of the mimic morph resembles the profile of ant larvae more than that of the alternative, genetically identical nonmimic morph. Furthermore, we show that, once in the brood chamber, mimic aphids suck on ant larva hemolymph. These results not only add aphids to the limited list of arthropods known to biosynthesize the cuticular chemicals of their deceived hosts to exploit their resources but describe a remarkable case of plastic aggressive mimicry. The present work adds a previously unidentified dimension to the classical textbook paradigm of aphid-ant relationships by showcasing a complex system at the evolutionary interface between cooperation and exploitation.

Brood space limitation of reproduction may explain growth after maturity in differently sized Daphnia species

Our aim was to determine if a limitation of reproduction by available brood chamber space could explain indeterminate growth in Daphnia, and what conditions might affect this. We evaluated the brood chamber volumes of the larger Daphnia longispina and smaller D. cucullata species during the first three clutches of eggs and compared this with the volume of eggs produced in low and high food conditions. We also examined whether these Daphnia retained significant levels of unused lipids in their bodies after reproduction, and if the volume of eggs deposited in brood chambers of different capacities corresponded to the maximal egg volume specified by a geometrical model. Our results revealed that, at least during the early clutches, both Daphnia species produced as manyeggs as permitted by the available brood chamber volume in both high and low food conditions, thus indicating that “brood space constraint” can explain the simultaneous investment in growth and reproduction. The detailed evaluation of this hypothesis required not only brood chamber volume assessment, but also consideration of the effect of egg size on packing efficiency, especially in low food conditions and for the smaller Daphnia species. The significant lipid reserves detected in the Daphnia body after reproduction also support the limitation of reproduction by brood chamber volume, particularly in larger species.

Vpliv krmljenja čebeljih družin na njihovo živalnost in pristnost medu

For the natural development of bee colonies, there is the need for appropriate nutrition. Lack of natural honey flow must be supplemented by feeding bee colonies with sugar syrups or candy paste. This supplementary feeding encourages brood breeding and forage activity, whereby stronger colonies collect more honey. Sugar syrups can cause honey adulteration, which is more frequent with the reversing of the brood combs with the bee food, with the combs moved from the brood chamber to the upper chamber. Authentication of honey from the standpoint of the presence of sugar syrup is very complex, because there is no single method by which honey adulteration can be reliably confirmed. Feeding the colonies in spring should result in stronger colonies and hence the collection of more honey in the brood chambers. The objective of the present study was to determine whether this has effects also on honey authenticity, and to discover a simple method for detection of honey adulteration. The colonies were fed with candy paste that had added yeast and blue dye, to provide markers for detection of honey adulteration. The strength of the colonies and quantity of honey in the brood chambers were monitored. The results of the analysis of stable isotope and activity of foreign enzymes were compared with the results of yeast quantity and colour of the honey (absorbance, L*, a*, b* parameters). Detection of yeast in the honey samples and presence of colour as a consequence of added dye appear to be appropriate methods to follow honey adulteration, and further studies are ongoing.

Waste Management in the Stingless Bee Melipona beecheii Bennett (Hymenoptera: Apidae)

Waste management is important in insect societies because waste can be hazardous to adults, brood and food stores. The general organization of waste management and the influence of task partitioning, division of labor and age polyethism on waste processing were studied in three colonies of the tropical American stingless bee Melipona beecheii Bennett in Yucatan, Mexico. Waste generated in the colony (feces, old brood cells, cocoons, dead adults and brood) was collected by workers throughout the nest and taken to specific waste dumps within the nest. During the day, workers based at the waste dumps formed waste pellets, which they directly transferred in 93% of cases, to other workers who subsequently removed them from the nest. This is an example of task partitioning and is hypothesized to improve nest hygiene as has been found in leafcutting ants, Atta. To investigate division of labor and age polyethism we marked a cohort of 144 emerging workers. Workers forming waste pellets were on average 31.2±6.5 days old (±SD, N= 40, range of 18-45 days). The life span of M. beecheii workers was 49.0±14.0 days (N= 144). There was no difference in the life span of workers who formed (52.2±11.6 days, N= 40) or did not form (49.9±11.5 days, N= 97) waste pellets, suggesting that waste work did not increase mortality. Although waste was probably not hazardous to adults and brood, because the dumps are located outside the brood chamber, its presence inside the nests can attract phorid flies and predators, which can harm the colony.

Bee Health in Apiaries in the Paraíba Valley, São Paulo State, Southeast Brazil

Bee health is a growing global concern due to phenomena with as yet undefined causes, such as the sudden population decline of colonies that has been observed in apiaries in many countries, recently including Brazil. The main objective of this study was to assess the presence and/or prevalence of pathogens that afflict Africanized Apis mellifera bees in the Paraiba Valley region of Sao Paulo state, Brazil. Three sampling periods were established: period 1 – August and September 2009 (winter/start of spring); period 2 – December 2009 and January 2010 (summer); and period 3 – April and May 2010 (autumn). Different types of samples were collected from the brood chamber, besides foragers at the entrance of the hive, in order to evaluate the presence and prevalence of Paenibacillus larvae, Varroa destructor and Nosema sp. The results indicated that the intensity of infection by Nosema ceranae and infestation rates of V. destructor in the hives are low, with no detection of P. larvae spores in the samples analyzed. The prevalence of N. ceranae and V. destructor was high, at respective values of 85.2 and 95.7%. These results demonstrate that although mites and microsporidia are widespread in the region’s colonies, the Africanized bees are apparently tolerant of these parasites. However, the mechanisms related to defense against pathogens are not completely clear and monitoring and prophylactic measures are essential to maintain the health of bee colonies.

Effects of low salinity on adult behavior and larval performance in the intertidal gastropod Crepipatella peruviana (Calyptraeidae).

Shallow-water coastal areas suffer frequent reductions in salinity due to heavy rains, potentially stressing the organisms found there, particularly the early stages of development (including pelagic larvae). Individual adults and newly hatched larvae of the gastropod Crepipatella peruviana were exposed to different levels of salinity stress (32(control), 25, 20 or 15), to quantify the immediate effects of exposure to low salinities on adult and larval behavior and on the physiological performance of the larvae. For adults we recorded the threshold salinity that initiates brood chamber isolation. For larvae, we measured the impact of reduced salinity on velar surface area, velum activity, swimming velocity, clearance rate (CR), oxygen consumption (OCR), and mortality (LC50); we also documented the impact of salinity discontinuities on the vertical distribution of veliger larvae in the water column. The results indicate that adults will completely isolate themselves from the external environment by clamping firmly against the substrate at salinities ≤24. Moreover, the newly hatched larvae showed increased mortality at lower salinities, while survivors showed decreased velum activity, decreased exposed velum surface area, and decreased mean swimming velocity. The clearance rates and oxygen consumption rates of stressed larvae were significantly lower than those of control individuals. Finally, salinity discontinuities affected the vertical distribution of larvae in the water column. Although adults can protect their embryos from low salinity stress until hatching, salinities <24 clearly affect survival, physiology and behavior in early larval life, which will substantially affect the fitness of the species under declining ambient salinities.