Pesticide exposure in honey bees results in increased levels of the gut pathogen <Emphasis Type="Italic">Nosema</Emphasis>

Global pollinator declines have been attributed to habitat destruction, pesticide use, and climate change or some combination of these factors, and managed honey bees, Apis mellifera, are part of worldwide pollinator declines. Here we exposed honey bee colonies during three brood generations to sub-lethal doses of a widely used pesticide, imidacloprid, and then subsequently challenged newly emerged bees with the gut parasite, Nosema spp. The pesticide dosages used were below levels demonstrated to cause effects on longevity or foraging in adult honey bees. Nosema infections increased significantly in the bees from pesticide-treated hives when compared to bees from control hives demonstrating an indirect effect of pesticides on pathogen growth in honey bees. We clearly demonstrate an increase in pathogen growth within individual bees reared in colonies exposed to one of the most widely used pesticides worldwide, imidacloprid, at below levels considered harmful to bees. The finding that individual bees with undetectable levels of the target pesticide, after being reared in a sub-lethal pesticide environment within the colony, had higher Nosema is significant. Interactions between pesticides and pathogens could be a major contributor to increased mortality of honey bee colonies, including colony collapse disorder, and other pollinator declines worldwide.     

The ‘I see you’ prey–predator signal of Apis cerana is innate

An ‘I see you’ (ISY) prey–predator signal can co-evolve when such a signal benefits both prey and predator. The prey benefits if, by producing the signal, the predator is likely to break off an attack. The predator benefits if it is informed by the signal that the prey is aware of its presence and can break off what is likely to be an unsuccessful and potentially costly hunt. Because the signal and response co-evolve in two species, the behaviour underlying an ISY signal is expected to have a strong genetic component and cannot be entirely learned. An example of an ISY signal is the ‘shimmering’ behaviour performed by Asian hive bee workers in the presence of their predator Vespa velutina. To test the prediction that bee–hornet signalling is heritable, we let honey bee workers of two species emerge in an incubator so that they had never been exposed to V. velutina. In Apis cerana, the shimmering response developed 48 h post-emergence, was strong after 72 h and increased further over 2 weeks. In contrast, A. mellifera, which has evolved in the absence of Asian hornets, did not produce the shimmering signal. In control tests, A. cerana workers exposed to a non-threatening butterfly did not respond with the shimmering signal.     

Large pollen loads of a South African asclepiad do not interfere with the foraging behaviour or efficiency of pollinating honey bees

The pollen of asclepiads (Asclepiadoideae, Apocynaceae) and most orchids (Orchidaceae) are packaged as large aggregations known as pollinaria that are removed as entire units by pollinators. In some instances, individual pollinators may accumulate large loads of these pollinaria. We found that the primary pollinator of Cynanchum ellipticum (Apocynaceae—Asclepiadoideae), the honey bee Apis mellifera, accumulate very large agglomerations of pollinaria on their mouthparts when foraging on this species. We tested whether large pollinarium loads negatively affected the foraging behaviour and foraging efficiency of honey bees by slowing foraging speeds or causing honey bees to visit fewer flowers, and found no evidence to suggest that large pollinarium loads altered foraging behaviour. C. ellipticum displayed consistently high levels of pollination success and pollen transfer efficiency (PTE). This may be a consequence of efficiently loading large numbers of pollinaria onto pollinators even when primary points of attachment on pollinators are already occupied and doing so in a manner that does not impact the foraging behaviour of pollinating insects.     

Effects of nitrogen dioxide and its acid mist on reactive oxygen species production and antioxidant enzyme activity in Arabidopsis plants

Nitrogen dioxide (NO₂) is one of the most common and harmful air pollutants. To analyze the response of plants to NO₂ stress, we investigated the morphological change, reactive oxygen species (ROS) production and antioxidant enzyme activity in Arabidopsis thaliana (Col-0) exposed to 1.7, 4, 8.5, and 18.8 mg/m³ NO₂. The results indicate that NO₂ exposure affected plant growth and chlorophyll (Chl) content, and increased oxygen free radical (O₂⁻) production rate in Arabidopsis shoots. Furthermore, NO₂ elevated the levels of lipid peroxidation and protein oxidation, accompanied by the induction of antioxidant enzyme activities and change of ascorbate (AsA) and glutathione (GSH) contents. Following this, we mimicked nitric acid mist under experimental conditions, and confirmed the antioxidant mechanism of the plant to the stress. Our results imply that NO₂ and its acid mist caused pollution risk to plant systems. During the process, increased ROS acted as a signal to induce a defense response, and antioxidant status played an important role in plant protection against NO₂/nitric acid mist-caused oxidative damage.     

Genetic diversity affects colony survivorship in commercial honey bee colonies

Honey bee (Apis mellifera) queens mate with unusually high numbers of males (average of approximately 12 drones), although there is much variation among queens. One main consequence of such extreme polyandry is an increased diversity of worker genotypes within a colony, which has been shown empirically to confer significant adaptive advantages that result in higher colony productivity and survival. Moreover, honey bees are the primary insect pollinators used in modern commercial production agriculture, and their populations have been in decline worldwide. Here, we compare the mating frequencies of queens, and therefore, intracolony genetic diversity, in three commercial beekeeping operations to determine how they correlate with various measures of colony health and productivity, particularly the likelihood of queen supersedure and colony survival in functional, intensively managed beehives. We found the average effective paternity frequency (m _e ) of this population of honey bee queens to be 13.6?±?6.76, which was not significantly different between colonies that superseded their queen and those that did not. However, colonies that were less genetically diverse (headed by queens with m _e ?≤?7.0) were 2.86 times more likely to die by the end of the study when compared to colonies that were more genetically diverse (headed by queens with m _e ?>?7.0). The stark contrast in colony survival based on increased genetic diversity suggests that there are important tangible benefits of increased queen mating number in managed honey bees, although the exact mechanism(s) that govern these benefits have not been fully elucidated.     

Heat Shielding: A Novel Method of Colonial Thermoregulation in Honey Bees

 Honey bees, Apis mellifera, maintain constant colony temperatures throughout the year. Honey bees fan their wings to cool the colony, and often spread fluid in conjunction with this behavior to induce evaporative cooling. We present an additional, previously undescribed mechanism used by the honey bee to maintain constant colony temperature in response to localized temperature increases. Worker bees shield the comb from external heat sources by positioning themselves on hot interior regions of the hive's walls. Although honey comb and brood comb were both shielded, the temperature-sensitive brood received a greater number of heat shielders and was thus better protected from overheating. Heat shielding appears to be a context-dependent adaptive behavior performed by worker bees who would previously have been considered "unemployed." Received: 16 November 1998 / Accepted in revised form: 31 March 1999     

Nickel accumulation and its effect on biomass, protein content and antioxidative enzymes in roots and leaves of watercress （Nasturtium officinale R. Br.）

In order to understand its response towards nickel stress, watercress (Nasturtium o cinale R. Br.) was exposed to nickel (1–25 mg/L) for 1, 3, 5 and 7 days. The accumulation and translocation of nickel were determined and the influence of nickel on biomass, protein content and enzymatic antioxidants was examined for both roots and leaves. It was determined that N. o cinale could accumulate appreciable amounts of Ni in both roots and leaves. Nickel accumulated particularly in the roots of plants. Biomass increased at low nickel concentrations but certain measurable change was not found at high concentrations. Under stress conditions the antioxidant enzymes were up-regulated compared to control. An increase in protein content and enzyme activities was observed at moderate exposure conditions followed by a decline at both roots and leaves. The maximum enzyme activities were observed at di erent exposure conditions. Our results showed that N. o cinale had the capacity to overcome nickel-induced stress especially at moderate nickel exposure. Therefore, N. o cinale may be used as a phytoremediator in moderately polluted aquatic ecosystems.     

Dietary antioxidants, lipid peroxidation and plumage colouration in nestling blue tits Cyanistes caeruleus

Carotenoid pigments are responsible for many of the red, yellow and orange plumage and integument traits seen in birds. One idea suggests that since carotenoids can act as antioxidants, carotenoid-mediated colouration may reveal an individual's ability to resist oxidative damage. In fact, there is currently very little information on the effects of most dietary-acquired antioxidants on oxidative stress in wild birds. Here, we assessed the impacts on oxidative damage, plasma antioxidants, growth and plumage colouration after supplementing nestling blue tits Cyanistes caeruleus with one of three diets; control, carotenoid treatment or α-tocopherol treatment. Oxidative damage was assessed by HPLC analysis of plasma levels of malondialdehyde (MDA), a by-product of lipid peroxidation. Contrary to predictions, we found no differences in oxidative damage, plumage colouration or growth rate between treatment groups. Although plasma lutein concentrations were significantly raised in carotenoid-fed chicks, α-tocopherol treatment had no effect on concentrations of plasma α-tocopherol compared with controls. Interestingly, we found that faster growing chicks had higher levels of oxidative damage than slower growing birds, independent of treatment, body mass and condition at fledging. Moreover, the chromatic signal of the chest plumage of birds was positively correlated with levels of MDA but not plasma antioxidant concentrations: more colourful nestlings had higher oxidative damage than less colourful individuals. Thus, increased carotenoid-mediated plumage does not reveal resistance to oxidative damage for nestling blue tits, but may indicate costs paid, in terms of oxidative damage. Our results indicate that the trade-offs between competing physiological systems for dietary antioxidants are likely to be complex in rapidly developing birds. Moreover, interpreting the biological relevance of different biomarkers of antioxidant status represents a challenge for evolutionary ecology.     

Physiological responses and detoxific mechanisms to Pb, Zn, Cu and Cd in young seedlings of Paulownia fortunei

Paulownia fortunei has been successfully used in the phytoremediation of many Pb/Zn mine tailings. However, seed germination and young seedlings of P. fortunei rarely occurred in these mine tailings. The physiological responses and detoxific mechanisms of P. fortunei young seedling to Pb, Zn, Cu and Cd stress were investigated. The germinated rate, shoot length, chlorophyll and carotenoid contents in leaves of young seedlings had a great reduction under Zn and Cu treatments, but had little decrease under Pb and Cd treatments. The production rate of O₂^•−, H₂O₂ and malondialdehyde (MDA) contents significantly increased in response to added Zn and Cu indicating great oxidative stress for young seedlings, but they had no significant change to added Pb and Cd. Young seedlings had effective detoxific mechanism to Pb and Cd, as antioxidant enzymes activities, phytochelatins (PCs-SH) and proline contents increased with increasing rates of added Pb and Cd. However, young seedlings had un-effective detoxific mechanisms to Zn and Cu stress. Results revealed the heavy metals (such as Cu) that present at low concentrations in mine tailings may be major constraint for the survival of young seedlings.     

Piriformospora indica confers cadmium tolerance in Nicotiana tabacum

Piriformospora indica, a root-colonizing endophytic fungus of Sebacinales, promotes plant growth and confers resistance against biotic and abiotic stresses. In order to confirm the influence of P. indica on growth, proline, malondialdehyde (MDA), chlorophyll, and cadmium (Cd) amounts in Nicotiana tabacum under Cd stress, hydroponics, pot and field trials were conducted. The results showed that P. indica can store Cd in plant roots and reduce leaf Cd content, reduce the concentration of MDA, and increase the proline and chlorophyll content and the activities of catalase, peroxidase, and superoxide dismutase under hydroponic Cd stress. RT-PCR analysis showed that the relative expression level of genes Gsh2, TaPCS1, oas1, GPX, and Hsp70 in colonized plants was 4.3, 1.4, 2.9, 1.7, and 6.9 fold higher than in un-colonized plants respectively. Cd exposure significantly reduced un-colonized plants'' agronomic traits compared to P. indica-colonized ones. Our results suggested that P. indica can sequester Cd in roots, so that much less cadmium was transported to leaves, and the increased concentrations of antioxidant enzymes, pigments and proline contents, as well as the higher expression of stress-related phytochelatin biosynthesis genes in P. indica-inoculated plants, may also serve to protect N. tabacum plants against oxidative damage, enhancing Cd tolerance.     

邻苯二甲酸二丁酯诱导小鼠神经行为学改变及相关生理生化指标分析

探究邻苯二甲酸二丁酯诱导小鼠神经行为学改变及与细胞外调节蛋白激酶(ERK1/2)通路相关蛋白的关联.雄性KM小鼠36只,随机分成4组:生理盐水组、50 mg·kg~(-1)·d~(-1) DBP组、50 mg·kg~(-1)·d~(-1)维生素E(VE)组、DBP+VE组,连续灌胃处理28 d,观察Morris水迷宫结果,检测小鼠脑海马组织的氧化应激(活性氧(ROS)荧光强度、还原型谷胱甘肽(GSH)与丙二醛(MDA)含量)、脑源性神经营养因子(BDNF)、磷酸化cAMP反应元件结合蛋白(p-CREB)、caspase-3水平,Western blot分析ERK1/2及其磷酸化(p-ERK1/2)水平;HE、Nissl染色及Hoechst 33258荧光染色分析脑组织CA1区病理学变化.结果表明,与对照组比较,50 mg·kg~(-1)·d~(-1) DBP组小鼠的学习记忆下降,氧化应激、p-ERK1/2、caspase-3水平上升,BDNF、p-CREB表达下降,差异均有统计学意义(p0.05,p0.01);海马组织CA1病理学损伤及凋亡程度增加.给予抗氧化剂VE处理后,DBP+VE组小鼠的学习记忆上升,氧化应激、p-ERK1/2、caspase-3水平降低,BDNF、p-CREB表达上升,差异均有统计学意义(p0.05);海马组织CA1病理学损伤及凋亡程度降低.由此推测,DBP暴露导致小鼠海马组织CA1病理学损伤、神经元凋亡程度增加、学习记忆下降、其神经行为学改变可以通过添加VE得到缓解,相关生理生化指标测试表明ERK系统应激氧化性损伤机制参与介导了毒理学过程.     

NR途径参与调节SO2胁迫下拟南芥含硫抗氧化物水平

以拟南芥为材料,研究了硝酸还原酶(NR)途径在植株响应SO_2过程中对含硫抗氧化物水平的影响.结果发现,野生型拟南芥植株中谷胱甘肽过氧化物酶(GPX)和谷胱甘肽硫转移酶(GST)活性高于NR缺失突变体(nia1nia2);SO_2暴露后,野生型和突变体nia1nia2植株中半胱氨酸(Cys)和谷胱甘肽(GSH)含量显著增加,GPX和GST的活性诱导性增高,且野生型植株中上述4项指标的增幅高于突变体;拟南芥逆境响应基因GSTU24和GPX7在野生型和突变体nia1nia2中差异表达,且对SO_2的响应程度不同.研究表明,植株体内NR途径与拟南芥细胞中含硫抗氧化物水平有关,NR缺失影响逆境应答中含硫抗氧化物水平的提高及抗逆基因转录,说明NR途径参与调节了植株逆境响应中含硫抗氧化物的合成及相关氧化还原反应与解毒过程.     

Toxic effects of perfluorononanoic acid on the development of Zebrafish (Danio rerio) embryos

Perfluorononanoic acid (PFNA) is a nine-carbon perfluoroalkyl acid widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans and wildlife. There is a concern that this chemical might be a developmental toxicant and teratogen in various ecosystems. In the present study, the toxic effects of PFNA were evaluated in zebrafish (Danio rerio) embryos. One hour post-fertilization embryos were treated with 0, 25, 50, 100, 200, 300, 350, and 400 μmol/L PFNA for 96 hr in 6-well plates. Developmental phenotypes and hatching rates were observed and recorded. Nineteen genes related to oxidative stress and lipid metabolism were examined using Quantitative RT-PCR and confirmed by whole mount in situ hybridization (WISH). Results showed that PFNA delayed the development of zebrafish embryos, reduced the hatching rate, and caused ventricular edema and malformation of the spine. In addition, the amount of reactive oxygen species in the embryo bodies increased significantly after exposure to PFNA compared with that of the control group. The Quantitative RT-PCR and WISH experiments demonstrated that mRNA expression of the lfabp and ucp2 genes increased significantly while that of sod1 and mt-nd1 decreased significantly after PFNA exposure. The mRNA expression levels of gpx1 and mt-atp6 decreased significantly in the high concentration group. However, the mRNA expression levels of both ppara and pparg did not show any significant variation after exposure. These findings suggest that PFNA affected the development of zebrafish embryos at relatively low concentrations.     

3种胡枝子抗氧化酶和渗透调节物质对干旱和增强UV-B辐射的动态响应

为探究干旱和增强UV-B辐射对3种胡枝子(美丽胡枝子、二色胡枝子、牛枝子)抗氧化酶活性和渗透调节物质的影响,采用盆栽试验,研究了正常生长(CK)、干旱胁迫(D)、增强UV-B辐射(UV-B)及二者复合胁迫(D+UV-B)下各植物叶片中抗氧化酶活性和渗透调节物质的动态响应.结果表明:3种处理下各植物叶片中SOD活性较对照值均呈先下降后上升趋势,且与单因子处理相比,牛枝子和二色胡枝子在复合胁迫下其SOD活性更高;增强UV-B辐射和复合处理下POD活性随胁迫时间总体呈先上升后下降趋势;CAT活性在干旱和复合胁迫下高于对照值,而在增强UV-B辐射下较对照值有所下降.3种植物叶片渗透调节物质含量(可溶性蛋白、可溶性糖、游离脯氨酸)在各处理中较对照值均有所增加,并呈先上升后下降趋势,但种间差异显著.3种处理均导致各植物叶片内MDA含量不同程度的升高,升高幅度的大小依次为牛枝子＞美丽胡枝子＞二色胡枝子.通过隶属函数法综合评价得出,复合处理下二色胡枝子和美丽胡枝子具有较强的抗氧化能力,牛枝子抗氧化能力较弱.综上,与干旱胁迫和增强UV-B辐射相比,复合胁迫对植物的伤害程度更小,说明干旱与增强UV-B辐射可以相互缓减彼此对植物的伤害,且美丽胡枝子和二色胡枝子在抵抗干旱胁迫和增强UV-B辐射方面具有一定优势.     

Vasculature of the hive: heat dissipation in the honey bee (Apis mellifera) hive

Eusocial insects are distinguished by their elaborate cooperative behavior and are sometimes defined as superorganisms. As a nest-bound superorganism, individuals work together to maintain favorable nest conditions. Residing in temperate environments, honey bees (Apis mellifera) work especially hard to maintain brood comb temperature between 32 and 36 °C. Heat shielding is a social homeostatic mechanism employed to combat local heat stress. Workers press the ventral side of their bodies against heated surfaces, absorb heat, and thus protect developing brood. While the absorption of heat has been characterized, the dissipation of absorbed heat has not. Our study characterized both how effectively worker bees absorb heat during heat shielding, and where worker bees dissipate absorbed heat. Hives were experimentally heated for 15 min during which internal temperatures and heat shielder counts were taken. Once the heat source was removed, hives were photographed with a thermal imaging camera for 15 min. Thermal images allowed for spatial tracking of heat flow as cooling occurred. Data indicate that honey bee workers collectively minimize heat gain during heating and accelerate heat loss during cooling. Thermal images show that heated areas temporarily increase in size in all directions and then rapidly decrease to safe levels (<37 °C). As such, heat shielding is reminiscent of bioheat removal via the cardiovascular system of mammals.     

毒死蜱对斑马鱼胚胎氧化应激效应研究

以斑马鱼为模型,研究了毒死蜱对斑马鱼胚胎形态学的影响,及其对胚胎的氧化应激和氧化损伤作用.将斑马鱼胚胎暴露在梯度浓度的毒死蜱溶液中96h后,发现毒死蜱会造成斑马鱼胚胎严重畸形甚至死亡,其96h半致死浓度为1.18mg/L.对氧化应激相关基因的表达及抗氧化酶活性和丙二醛(MDA)含量进行检测,结果表明,在毒死蜱胁迫下抗氧化酶(SOD、CAT)活性降低,并且其编码基因(Cu/Zn-sod、Mn-sod、cat)的表达受到抑制;但在低浓度毒死蜱胁迫下,抗氧化酶活性并没有受到显著影响,而抗氧化酶基因的表达对毒死蜱更加敏感.毒死蜱能引起gstp2的表达上调,但GST活性与gstp2的表达变化并不一致.处理组胚胎中nrf2表达上调,从而上调抗氧化蛋白和II相解毒酶基因的表达.毒死蜱胁迫下,基因ucp2、cox1表达下调,能够减少呼吸链ROS的产生.同时基因bcl2表达下调,表明凋亡的平衡受到破坏.毒死蜱处理组中MDA含量显著升高,说明毒死蜱能造成斑马鱼胚胎氧化损伤.     

Honeybee glucose oxidase—its expression in honeybee workers and comparative analyses of its content and H2O2-mediated antibacterial activity in natural honeys

Antibacterial properties of honey largely depend on the accumulation of hydrogen peroxide (H₂O₂), which is generated by glucose oxidase (GOX)-mediated conversion of glucose in diluted honey. However, honeys exhibit considerable variation in their antibacterial activity. Therefore, the aim of the study was to identify the mechanism behind the variation in this activity and in the H₂O₂ content in honeys associated with the role of GOX in this process. Immunoblots and in situ hybridization analyses demonstrated that gox is solely expressed in the hypopharyngeal glands of worker bees performing various tasks and not in other glands or tissues. Real-time PCR with reference genes selected for worker heads shows that the gox expression progressively increases with ageing of the youngest bees and nurses and reached the highest values in processor bees. Immunoblot analysis of honey samples revealed that GOX is a regular honey component but its content significantly varied among honeys. Neither botanical source nor geographical origin of honeys affected the level of GOX suggesting that some other factors such as honeybee nutrition and/or genetic/epigenetic factors may take part in the observed variation. A strong correlation was found between the content of GOX and the level of generated H₂O₂ in honeys except honeydew honeys. Total antibacterial activity of most honey samples against Pseudomonas aeruginosa isolate significantly correlated with the H₂O₂ content. These results demonstrate that the level of GOX can significantly affect the total antibacterial activity of honey. They also support an idea that breeding of novel honeybee lines expressing higher amounts of GOX could help to increase the antibacterial efficacy of the hypopharyngeal gland secretion that could have positive influence on a resistance of colonies against bacterial pathogens.