Julia Horwood

Neonicotinoid Residues in Wildflowers, a Potential Route of Chronic Exposure for Bees

In recent years, an intense debate about the environmental risks posed by neonicotinoids, a group of widely used, neurotoxic insecticides, has been joined. When these systemic compounds are applied to seeds, low concentrations are subsequently found in the nectar and pollen of the crop, which are then collected and consumed by bees. Here we demonstrate that the current focus on exposure to pesticides via the crop overlooks an important factor: throughout spring and summer, mixtures of neonicotinoids are also found in the pollen and nectar of wildflowers growing in arable field margins, at concentrations that are sometimes even higher than those found in the crop. Indeed, the large majority (97%) of neonicotinoids brought back in pollen to honey bee hives in arable landscapes was from wildflowers, not crops. Both previous and ongoing field studies have been based on the premise that exposure to neonicotinoids would occur only during the blooming period of flowering crops and that it may be diluted by bees also foraging on untreated wildflowers. Here, we show that exposure is likely to be higher and more prolonged than currently recognized because of widespread contamination of wild plants growing near treated crops.

Bioassay-directed identification of novel antiandrogenic compounds in bile of fish exposed to wastewater effluents

The widespread occurrence of feminized male fish downstream of some UK Wastewater Treatment Works (WwTWs) has been associated with exposure to estrogenic and potentially antiandrogenic (AA) contaminants in the effluents. In this study, profiling of AA contaminants in WwTW effluents and fish was conducted using HPLC in combination with in vitro androgen receptor transcription screens. Analysis of extracts of wastewater effluents revealed complex profiles of AA activity comprising 21-53 HPLC fractions. Structures of bioavailable antiandrogens were identified by exposing rainbow trout to a WwTW effluent and profiling the bile for AA activity using yeast (anti-YAS) and mammalian-based (AR-CALUX) androgen receptor transcription screens. The predominant fractions with AA activity in both androgen receptor screens contained the germicides chlorophene and triclosan, and together these contaminants accounted for 51% of the total anti-YAS activity in the fish bile. Other AA compounds identified in bile included chloroxylenol, dichlorophene, resin acids, napthols, oxybenzone, 4-nonylphenol, and bisphenol A. Pure standards of these compounds were active in the androgen receptor screens at potencies relative to flutamide of between 0.1 and 13.0. Thus, we have identified, for the first time, a diverse range of AA chemicals in WwTWs that are bioavailable to fish and which need to be assessed for their risk to the reproductive health of these organisms and other aquatic biota.

Fundamental differences in the optical structure of the eyes of nocturnal and diurnal mosquitoes

Abstract We have studied the anatomy and optics of the eyes of a range of mosquito species from the wholly dark-active blood-feeding Anopheles gambiae to the diurnal plant-feeder Toxorhynchites brevipalpis. Consistent with studies by Satô in the 1950s, we find that dark-active and crepuscular species have short fused rhabdoms with a conical construction. This maximises the amount of light the rhabdoms receive from the almost hemispherical wide-aperture lenses. Toxorhynchites, on the other hand, has long narrow rhabdomeres that are separated from each other over their entire length, and so resemble the open rhabdoms of advanced flies (Brachycera and Cyclorrhapha). These findings are confirmed by studies of the pseudopupil, whose form indicates the layout of the rhabdomere tips in the focal plane of each ommatidial lens. In anopheline species the pseudopupil is a single undivided ellipse, indicating a fused rhabdom structure, whereas in Toxorhynchites there is a ring of six outer elements surrounding a central one. This means that each rhabdomere views a separate direction in space, and our measurements indicate that, as in higher Diptera, adjacent rhabdomeres share their fields of view with one of the rhabdomeres in the immediately adjacent ommatidia. This in turn means that in the diurnal type of mosquito eye there is a basis for neural superposition, but the fused construction of anopheline rhabdoms precludes this. The Aedes species studied were similar to Anopheles but with lenses of less extreme aperture, and Sabethes cyaneus, a diurnal blood-feeder, was intermediate in structure, with fused conical rhabdoms in the centre of the eye and unfused rhabdomeres around the periphery.

5‐HT2C receptor activation inhibits appetitive and consummatory components of feeding and increases brain c‐fos immunoreactivity in mice

5‐Hydroxytryptamine (5‐HT)2C and 5‐HT1B receptors are implicated in the inhibitory modulation of feeding behaviour. However, their respective, and possibly different, roles have not been clearly identified because of a lack of selective 5‐HT2C receptor agonists. Here, using the putative, selective 5‐HT2C receptor agonist VER23779, we show that its effects on feeding are fully reversed by pretreatment with a selective 5‐HT2C receptor antagonist, but unaffected by pretreatment with either a 5‐HT1B or a 5‐HT2A receptor antagonist. In mice eating a palatable mash, feeding ends earlier, inactivity is increased but the behavioural satiety sequence is preserved. In a second‐order schedule of reinforcement with an initial, non‐food‐reinforced appetitive phase, VER23779 produces a much greater relative reduction in appetitive responding than the 5‐HT1B receptor agonist CP‐94,253. Increased c‐fos immunoreactivity patterns following VER23779 also differ from those described for CP‐94,253, in particular showing strong activation of the basolateral amygdala. The different behavioural consequences of 5‐HT2C and 5‐HT1B receptor activation may relate to the patterns of c‐fos immunoreactivity. In particular, the basolateral amygdala may have a role in maintaining response in the appetitive phase of the second‐order schedule and also be susceptible to serotonergic modulation through activation of 5‐HT2C receptors.

Profiles and some initial identifications of (anti)androgenic compounds in fish exposed to wastewater treatment works effluents.

Exposure of fish to wastewater treatment works (WwTWs) effluents can result in reproductive anomalies consistent with exposure to estrogenic compounds. However, UK WwTWs effluents also contain compounds with androgen receptor activities which may contribute to reproductive dysfunction in fish. A toxicity identification and evaluation (TIE) approach was used to profile (anti)androgenic compounds in bile of fish exposed to two WwTWs effluents. Extracts of bile from exposed fish and effluent were fractionated by liquid chromatography and tested for (anti)androgenic activity using a yeast androgen receptor transcription screen (YAS). A number of bile fractions contained (anti)androgenic activity unique to the effluent-exposed fish. Some of these fractions contained di(chloromethyl)anthracene or dichlorophene, and these contaminants showed antagonistic activity in the YAS when tested as pure compounds. No androgenic activity was detected in the effluents, but TIE analysis of bile revealed a number of androgenic fractions which contained testosterone metabolites that were unique to effluent-exposed fish. This is the first work reported on the nature of some of the (anti)androgenic compounds that bioaccumulate in fish from WwTWs effluents and indicates that other contaminants, besides estrogenic substances, need to be considered for their potential to contribute to the disruption of reproductive system of fish in UK waters.

Disruption of the steroid metabolome in fish caused by exposure to the environmental estrogen 17α-ethinylestradiol.

Exposure to environmental estrogens such as 17alpha-ethinylestradiol (EE2) has been associated with feminization and a decline in fertility of male fish. To investigate the effect of estrogen exposure on steroid homeostasis, we exposed roach (Rutilus rutilus) to EE2 (1-29 ng/L) for 18 days and analyzed steroid profiles in bile and plasma using targeted analyses and in liver and gonadal tissues using mass spectrometry metabolite profiling techniques (metabolomics). Exposure to EE2 resulted in a concentration dependent reduction of estrogens and androgens in bile and plasma of both male and female fish. At 10 ngEE2/L, significant reductions in concentrations of hydroxyprogesterone, androstenedione, 11-hydroxyandrostenedione, and 11-ketotestosterone were detected in the testes metabolome, indicating disruption of steroid biosynthesis upstream of androgen metabolism. Estrogen exposure also resulted in increased biosynthesis of cortisol and cortisone in testes and ovaries, respectively, but did not alter glucocorticoid concentrations in the liver or plasma. This first report on the effect of EE2 exposure on the steroid metabolome in fish tissues suggests that both sex steroid and glucocorticoid pathways are one of the primary targets of estrogen exposure in fish gonads and provides further insights into the mode of action of this endocrine disrupting chemical.

Optics of the ultraviolet reflecting scales of a jumping spider

The jumping spider Cosmophasis umbratica from Singapore is strongly sexually dimorphic. The males, but not the females, reflect ultraviolet as well as green–orange light. The scales responsible for this are composed of a chitin–air–chitin sandwich in which the chitin layers are three-quarters of a wavelength thick and the air gap a quarter wavelength (where λ=600 nm, the peak wavelength of the principal reflection maximum). It is shown that this configuration produces a second reflectance peak at approximately 385 nm, accounting for the observed reflection in the ultraviolet. Other scales have a similar thickness of chitin but lack the air gap and thus produce a dull purple reflection. This novel mechanism provides the spiders with two colour signals, both of which are important in mating displays.

Neuroarchitecture of the color and polarization vision system of the Stomatopod haptosquilla

The apposition compound eyes of stomatopod crustaceans contain a morphologically distinct eye region specialized for color and polarization vision, called the mid‐band. In two stomatopod superfamilies, the mid‐band is constructed from six rows of enlarged ommatidia containing multiple photoreceptor classes for spectral and polarization vision. The aim of this study was to begin to analyze the underlying neuroarchitecture, the design of which might reveal clues how the visual system interprets and communicates to deeper levels of the brain the multiple channels of information supplied by the retina. Reduced silver methods were used to investigate the axon pathways from different retinal regions to the lamina ganglionaris and from there to the medulla externa, the medulla interna, and the medulla terminalis. A swollen band of neuropil—here termed the accessory lobe—projects across the equator of the lamina ganglionaris, the medulla externa, and the medulla interna and represents, structurally, the retinas mid‐band. Serial semithin and ultrathin resin sections were used to reconstruct the projection of photoreceptor axons from the retina to the lamina ganglionaris. The eight axons originating from one ommatidium project to the same lamina cartridge. Seven short visual fibers end at two distinct levels in each lamina cartridge, thus geometrically separating the two channels of polarization and spectral information. The eighth visual fiber runs axially through the cartridge and terminates in the medulla externa. We conclude that spatial, color, and polarization information is divided into three parallel data streams from the retina to the central nervous system. J. Comp. Neurol. 467:326–342, 2003.

Mosquito eye design: conical rhabdoms are matched to wide aperture lenses

Night–active mosquitoes have photopigment–containing rhabdoms that take the form of a wide hollow cone, with the distal apex at the focus of the ommatidial lens. It is shown geometrically, and with the aid of a gelatin model, that such a structure can trap light over an angle of up to 60°, roughly equal to the width of the cone of light provided by the lens. This gives a gain in light–gathering power of up to nine times that of the more usual cylinder–shaped rhabdom with an acceptance angle of less than 25°.

Infection by a foliar endophyte elicits novel arabidopside-based plant defence reactions in its host,Cirsium arvense

Endophytic fungi live asymptomatically within plants. They are usually regarded as nonpathogenic or even mutualistic, but whether plants respond antagonistically to their presence remains unclear, particularly in the little-studied associations between endophytes and nongraminoid herbaceous plants. We investigated the effects of the endophyte Chaetomium cochlioides on leaf chemistry in Cirsium arvense. Plants were sprayed with spores; leaf material from both subsequent new growth and the sprayed leaves was analysed 2 wk later. Infection frequency was 91% and 63% for sprayed and new growth, respectively, indicating that C. cochlioides rapidly infects new foliage. Metabolomic analyses revealed marked changes in leaf chemistry with infection, especially in new growth. Changes in several novel oxylipin metabolites were detected, including arabidopsides reported here for the first time in a plant species other than Arabidopsis thaliana, and a jasmonate-containing galactolipid. The production of these metabolites in response to endophyte presence, particularly in newly infected foliage, suggests that endophytes elicit similar chemical responses in plants to those usually produced following wounding, herbivory and pathogen invasion. Whether endophytes benefit their hosts may depend on a complex series of chemically mediated interactions between the plant, the endophyte, other microbial colonists and natural enemies.