Social Modulation of Stress Reactivity and Learning in Young Worker Honey Bees

Alarm pheromone and its major component isopentylacetate induce stress-like responses in forager honey bees, impairing their ability to associate odors with a food reward. We investigated whether isopentylacetate exposure decreases appetitive learning also in young worker bees. While isopentylacetate-induced learning deficits were observed in guards and foragers collected from a queen-right colony, learning impairments resulting from exposure to this pheromone could not be detected in bees cleaning cells. As cell cleaners are generally among the youngest workers in the colony, effects of isopentylacetate on learning behavior were examined further using bees of known age. Adult workers were maintained under laboratory conditions from the time of adult emergence. Fifty percent of the bees were exposed to queen mandibular pheromone during this period, whereas control bees were not exposed to this pheromone. Isopentylacetate-induced learning impairments were apparent in young (less than one week old) controls, but not in bees of the same age exposed to queen mandibular pheromone. This study reveals young worker bees can exhibit a stress-like response to alarm pheromone, but isopentylacetate-induced learning impairments in young bees are suppressed by queen mandibular pheromone. While isopentylacetate exposure reduced responses during associative learning (acquisition), it did not affect one-hour memory retrieval.     

Drone and Worker Brood Microclimates Are Regulated Differentially in Honey Bees,Apis mellifera

Background

Honey bee (Apis mellifera) drones and workers show differences in morphology, physiology, and behavior. Because the functions of drones are more related to colony reproduction, and those of workers relate to both survival and reproduction, we hypothesize that the microclimate for worker brood is more precisely regulated than that of drone brood.

Methodology/Principal Findings

We assessed temperature and relative humidity (RH) inside honey bee colonies for both drone and worker brood throughout the three-stage development period, using digital HOBO^® Data Loggers. The major findings of this study are that 1) both drone and worker castes show the highest temperature for eggs, followed by larvae and then pupae; 2) temperature in drones are maintained at higher precision (smaller variance) in drone eggs and larvae, but at a lower precision in pupae than the corresponding stages of workers; 3) RH regulation showed higher variance in drone than workers across all brood stages; and 4) RH regulation seems largely due to regulation by workers, as the contribution from empty honey combs are much smaller compared to that from adult workers.

Conclusions/Significance

We conclude that honey bee colonies maintain both temperature and humidity actively; that the microclimate for sealed drone brood is less precisely regulated than worker brood; and that combs with honey contribute very little to the increase of RH in honey bee colonies. These findings increase our understanding of microclimate regulation in honey bees and may have implications for beekeeping practices.     

Mapping Sleeping Bees within Their Nest: Spatial and Temporal Analysis of Worker Honey Bee Sleep

Patterns of behavior within societies have long been visualized and interpreted using maps. Mapping the occurrence of sleep across individuals within a society could offer clues as to functional aspects of sleep. In spite of this, a detailed spatial analysis of sleep has never been conducted on an invertebrate society. We introduce the concept of mapping sleep across an insect society, and provide an empirical example, mapping sleep patterns within colonies of European honey bees (Apis mellifera L.). Honey bees face variables such as temperature and position of resources within their colony''s nest that may impact their sleep. We mapped sleep behavior and temperature of worker bees and produced maps of their nest''s comb contents as the colony grew and contents changed. By following marked bees, we discovered that individuals slept in many locations, but bees of different worker castes slept in different areas of the nest relative to position of the brood and surrounding temperature. Older worker bees generally slept outside cells, closer to the perimeter of the nest, in colder regions, and away from uncapped brood. Younger worker bees generally slept inside cells and closer to the center of the nest, and spent more time asleep than awake when surrounded by uncapped brood. The average surface temperature of sleeping foragers was lower than the surface temperature of their surroundings, offering a possible indicator of sleep for this caste. We propose mechanisms that could generate caste-dependent sleep patterns and discuss functional significance of these patterns.     

Infestation of Japanese Native Honey Bees by Tracheal Mite and Virus from Non-native European Honey Bees in Japan

Invasion of alien species has been shown to cause detrimental effects on habitats of native species. Insect pollinators represent such examples; the introduction of commercial bumble bee species for crop pollination has resulted in competition for an ecological niche with native species, genetic disturbance caused by mating with native species, and pathogen spillover to native species. The European honey bee, Apis mellifera, was first introduced into Japan for apiculture in 1877, and queen bees have been imported from several countries for many years. However, its effects on Japanese native honey bee, Apis cerana japonica, have never been addressed. We thus conducted the survey of honey bee viruses and Acarapis mites using both A. mellifera and A. c. japonica colonies to examine their infestation in native and non-native honey bee species in Japan. Honey bee viruses, Deformed wing virus (DWV), Black queen cell virus (BQCV), Israeli acute paralysis virus (IAPV), and Sacbrood virus (SBV), were found in both A. mellifera and A. c. japonica colonies; however, the infection frequency of viruses in A. c. japonica was lower than that in A. mellifera colonies. Based on the phylogenies of DWV, BQCV, and SBV isolates from A. mellifera and A. c. japonica, DWV and BQCV may infect both honey bee species; meanwhile, SBV has a clear species barrier. For the first time in Japan, tracheal mite (Acarapis woodi) was specifically found in the dead honey bees from collapsing A. c. japonica colonies. This paper thus provides further evidence that tracheal-mite-infested honey bee colonies can die during cool winters with no other disease present. These results demonstrate the infestation of native honey bees by parasite and pathogens of non-native honey bees that are traded globally.     

A Comparative Study of Varroa Jacobsoni Reproduction in Worker Cells of Honey Bees (Apis Mellifera) in England and Africanized Bees in Yucatan, Mexico

The aim of this study was to investigate an underlying mechanism of the apparent tolerance of Africanized honey bees (AHB) to Varroa jacobsoni mites in Mexico. This was achieved by conducting the first detailed study into the mites' reproductive biology in AHB worker cells. The data was then compared directly with a similar study previously carried out on European honey bees (EHB) in the UK. A total of 1071 singly infested AHB worker cells were analyzed and compared with the data from 908 singly infested EHB worker cells. There was no significant difference between the number of mother mites dying in the cells (AHB = 2.0%, EHB = 1.8%); the mean number of eggs laid per mite (AHB = 4.86, EHB = 4.93); the number of mites producing no offspring (AHB = 12%, EHB = 9%); and developmental times of the offspring in worker cells of AHB and EHB. However, there was a major difference between the percentage of mother mites producing viable adult female offspring (AHB = 40%, EHB = 75%). This was caused by the increased rate of mite offspring mortality suffered by the first (male) and second (female) offspring in AHB worker cells. Therefore, only an average of 0.7 viable adult female offspring are produced per mite in AHB, compared to 1.0 in EHB.     

铅色水蛇繁殖生活史特征的年间变异

爬行动物繁殖生活史特征是动物进化生物学研究领域的重要内容,而国内有关卵胎生蛇类繁殖生活史特征的研究目前较为匮乏.本研究以我国南方地区常见的卵胎生蛇类——铅色水蛇(Hypsiscopus plumbea)为研究对象,在2013至2015年每年7月于温州地区捕获怀孕中后期水蛇母体,带回实验室,收集所产幼体,测量母体产后形态...     

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)

This video demonstrates novel techniques of RNA interference (RNAi) which downregulate two genes simultaneously in honey bees using double-stranded RNA (dsRNA) injections. It also presents a protocol of proboscis extension response (PER) assay for measuring gustatory perception.RNAi-mediated gene knockdown is an effective technique downregulating target gene expression. This technique is usually used for single gene manipulation, but it has limitations to detect interactions and joint effects between genes. In the first part of this video, we present two strategies to simultaneously knock down two genes (called double gene knockdown). We show both strategies are able to effectively suppress two genes, vitellogenin (vg) and ultraspiracle (usp), which are in a regulatory feedback loop. This double gene knockdown approach can be used to dissect interrelationships between genes and can be readily applied in different insect species.The second part of this video is a demonstration of proboscis extension response (PER) assay in honey bees after the treatment of double gene knockdown. The PER assay is a standard test for measuring gustatory perception in honey bees, which is a key predictor for how fast a honey bee''s behavioral maturation is. Greater gustatory perception of nest bees indicates increased behavioral development which is often associated with an earlier age at onset of foraging and foraging specialization in pollen. In addition, PER assay can be applied to identify metabolic states of satiation or hunger in honey bees. Finally, PER assay combined with pairing different odor stimuli for conditioning the bees is also widely used for learning and memory studies in honey bees.     

Predicting Gene Function from Uncontrolled Expression Variation among Individual Wild-Type Arabidopsis Plants

Gene expression profiling studies are usually performed on pooled samples grown under tightly controlled experimental conditions to suppress variability among individuals and increase experimental reproducibility. In addition, to mask unwanted residual effects, the samples are often subjected to relatively harsh treatments that are unrealistic in a natural context. Here, we show that expression variations among individual wild-type Arabidopsis thaliana plants grown under the same macroscopic growth conditions contain as much information on the underlying gene network structure as expression profiles of pooled plant samples under controlled experimental perturbations. We advocate the use of subtle uncontrolled variations in gene expression between individuals to uncover functional links between genes and unravel regulatory influences. As a case study, we use this approach to identify ILL6 as a new regulatory component of the jasmonate response pathway.     

Overwintering Is Associated with Reduced Expression of Immune Genes and Higher Susceptibility to Virus Infection in Honey Bees

The eusocial honey bee, Apis mellifera, has evolved remarkable abilities to survive extreme seasonal differences in temperature and availability of resources by dividing the worker caste into two groups that differ in physiology and lifespan: summer and winter bees. Most of the recent major losses of managed honey bee colonies occur during the winter, suggesting that winter bees may have compromised immune function and higher susceptibility to diseases. We tested this hypothesis by comparing the expression of eight immune genes and naturally occurring infection levels of deformed wing virus (DWV), one of the most widespread viruses in A. mellifera populations, between summer and winter bees. Possible interactions between immune response and physiological activity were tested by measuring the expression of vitellogenin and methyl farnesoate epoxidase, a gene coding for the last enzyme involved in juvenile hormone biosynthesis. Our data show that high DWV loads in winter bees correlate with reduced expression of genes involved in the cellular immune response and physiological activity and high expression of humoral immune genes involved in antibacterial defense compared with summer bees. This expression pattern could reflect evolutionary adaptations to resist bacterial pathogens and economize energy during the winter under a pathogen landscape with reduced risk of pathogenic viral infections. The outbreak of Varroa destructor infestation could have overcome these adaptations by promoting the transmission of viruses. Our results suggest that reduced cellular immune function during the winter may have increased honey bee’s susceptibility to DWV. These results contribute to our understanding of honey bee colony losses in temperate regions.     

山羊FGF5基因组织表达及其变异与羊绒性状的关联性分析

为了研究山羊FGF5基因的遗传变异和组织表达等分子特征及其变异对山羊羊绒性状的影响,本试验应用PCR-SSCP与测序相结合的方法检测FGF5基因第1外显子在陇东绒山羊、柴达木绒山羊和中卫山羊中的多态性,并分析该基因变异对羊绒性状的影响;采用RT-qPCR技术研究FGF5基因在陇东绒山羊和辽宁绒山羊组织中的表达.结果 表明,在山羊FGF5基因的第1外显子区检测到一个同义突变位点c.272C＞T,表现为MM、MN和NN3种基因型,其中NN型仅在陇东绒山羊中有极少量分布.3个山羊品种的优势基因型均为MM,优势等位基因均为M.c.272C＞T位点在3种山羊品种中均处于Hardy-Weinberg平衡状态,且均为低度多态.c.272C＞T位点与陇东绒山羊的羊绒性状关联性分析表明,该位点对羊绒纤维直径的影响显著.携带等位基因N的个体的羊绒纤维直径显著低于不含等位基因N的个体的(P＜0.05);基因型MN个体的羊绒纤维直径显著低于基因型MM个体的(P＜0.05).FGF5基因在陇东绒山羊和辽宁绒山羊的心、肝、脾、肺、肾、骨骼肌和皮肤中均有表达;在陇东绒山羊的心、脾、肺、肾和皮肤中的表达量显著或极显著高于辽宁绒山羊的.本研究结果提示,若以改善羊绒细度为选育目标,山羊FGF5基因可作为羊绒性状改良的候选基因.     

Modification of Gene Expression and Virulence Traits in Streptococcus mutans in Response to Carbohydrate Availability

The genetic and phenotypic responses of Streptococcus mutans, an organism that is strongly associated with the development of dental caries, to changes in carbohydrate availability were investigated. S. mutans UA159 or a derivative of UA159 lacking ManL, which is the EIIAB component (EIIAB^Man) of a glucose/mannose permease of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) and a dominant effector of catabolite repression, was grown in continuous culture to steady state under conditions of excess (100 mM) or limiting (10 mM) glucose. Microarrays using RNA from S. mutans UA159 revealed that 174 genes were differentially expressed in response to changes in carbohydrate availability (P < 0.001). Glucose-limited cells possessed higher PTS activity, could acidify the environment more rapidly and to a greater extent, and produced more ManL protein than cultures grown with excess glucose. Loss of ManL adversely affected carbohydrate transport and acid tolerance. Comparison of the histidine protein (HPr) in S. mutans UA159 and the manL deletion strain indicated that the differences in the behaviors of the strains were not due to major differences in HPr pools or HPr phosphorylation status. Therefore, carbohydrate availability alone can dramatically influence the expression of physiologic and biochemical pathways that contribute directly to the virulence of S. mutans, and ManL has a profound influence on this behavior.     

The Use of the Vibration Signal and Worker Piping to Influence Queen Behavior during Swarming in Honey Bees, Apis mellifera

We investigated worker regulation of queen activity during reproductive swarming by examining the rates at which workers performed vibration signals and piping on queens during the different stages of the swarming process. Worker–queen interactions were first examined inside observation hives during the 2–3 wk that preceded the issue of the swarm (pre‐swarming period) and then inside the swarm clusters during the period that preceded liftoff and relocation to a new nest site (post‐swarming period). Queen court size did not differ between the pre‐ and post‐swarming periods, but workers fed the queens less inside the swarm clusters. Workers performed vibration signals on the queens at increasing rates throughout the pre‐swarming period inside the natal nest, but rarely or never vibrated the queen inside the swarm. Piping was performed on the queens during both the pre‐ and post‐swarming periods and always reached a peak immediately before queen flight. During the final 2–4 h before swarm liftoff, queens were increasingly contacted by waggle dancers for nest sites, some of which piped the queen. The vibration signal may operate in a modulatory manner to gradually prepare the queen for flight from the natal nest, and the cumulative effects of the signal during the pre‐swarming period may make further vibrations on the queen unnecessary when inside the swarm cluster. In contrast, worker piping may function in a more immediate manner to trigger queen takeoff during both the pre‐ and post‐swarming periods. Workers that vibrate and pipe the queen tend to be older, foraging‐age bees. The regulation of queen activity during colony reproduction may therefore be controlled largely by workers that normally have little contact with queens, but help to formulate colony reproductive and movement decisions.     

MyoG基因对金华猪繁殖性状的影响

用PCR-RFLP方法对金华猪I系（109头）、II系（15头）、III系（25头）的MyoG基因的第二内含子内（PCR1- MspⅠ-RFLP）和3'-端（PCR2- MspⅠ-RFLP）位点进行了多态性分析。结果表明：PCR1- MspⅠ-RFLP位点有3种基因型（AA、AB、BB）,其基因型频率分别为0.1544,0.3826,0.4631;PCR2- MspⅠ-RFLP位点只有2种基因型（MM、MN）,其基因型频率分别为0.9953和0.0047。采用SPSS程序分析MyoG基因对金华猪繁殖性状的影响。结果表明：MyoG基因对金华母猪初胎的总产仔数、二胎的产活仔数影响显著（P<0.05）,而对其他胎次组合的总产仔数、产活仔数以及初生窝重没有显著影响（P<0.05）。