The COLOSS BEEBOOK Volume II: Standard methods for Apis mellifera pest and pathogen research

n recent years, declines of honey bee populations have received massive media attention worldwide, yet attempts to understand the causes have been hampered by a lack of standardisation of laboratory techniques. Published as a response to this, the COLOSS BEEBOOK is a unique collaborative venture involving 234 bee scientists from 34 countries, who have produced the definitive guide to how to carry out research on honey bees. It is hoped that these volumes will become the standards to be adopted by bee scientists worldwide. Volume II includes approximately 600 separate protocols dealing with the study of the pests and diseases of the honey bee, Apis mellifera. These cover epidemiology and surveying techniques, virus diseases, bacterial diseases such as European and American foulbrood, fungal and microsporidian diseases such as Nosema, mites such as Acarapis, Varroa and Tropilaelaps, and other pests such as the small hive beetle.

Differential Responses of Herbivores and Herbivory to Management in Temperate European Beech

Forest management not only affects biodiversity but also might alter ecosystem processes mediated by the organisms, i.e. herbivory the removal of plant biomass by plant-eating insects and other arthropod groups. Aiming at revealing general relationships between forest management and herbivory we investigated aboveground arthropod herbivory in 105 plots dominated by European beech in three different regions in Germany in the sun-exposed canopy of mature beech trees and on beech saplings in the understorey. We separately assessed damage by different guilds of herbivores, i.e. chewing, sucking and scraping herbivores, gall-forming insects and mites, and leaf-mining insects. We asked whether herbivory differs among different forest management regimes (unmanaged, uneven-aged managed, even-aged managed) and among age-classes within even-aged forests. We further tested for consistency of relationships between regions, strata and herbivore guilds. On average, almost 80 of beech leaves showed herbivory damage, and about 6 of leaf area was consumed. Chewing damage was most common, whereas leaf sucking and scraping damage were very rare. Damage was generally greater in the canopy than in the understorey, in particular for chewing and scraping damage, and the occurrence of mines. There was little difference in herbivory among differently managed forests and the effects of management on damage differed among regions, strata and damage types. Covariates such as wood volume, tree density and plant diversity weakly influenced herbivory, and effects differed between herbivory types. We conclude that despite of the relatively low number of species attacking beech; arthropod herbivory on beech is generally high. We further conclude that responses of herbivory to forest management are multifaceted and environmental factors such as forest structure variables affecting in particular microclimatic conditions are more likely to explain the variability in herbivory among beech forest plots.

The Honey Bee Pathosphere of Mongolia: European Viruses in Central Asia

Parasites and pathogens are apparent key factors for the detrimental health of managed European honey bee subspecies, Apis mellifera. Apicultural trade is arguably the main factor for the almost global distribution of most honey bee diseases, thereby increasing chances for multiple infestations/infections of regions, apiaries, colonies and even individual bees. This imposes difficulties to evaluate the effects of pathogens in isolation, thereby creating demand to survey remote areas. Here, we conducted the first comprehensive survey for 14 honey bee pathogens in Mongolia (N = 3 regions, N = 9 locations, N = 151 colonies), where honey bee colonies depend on humans to overwinter. In Mongolia, honey bees, Apis spp., are not native and colonies of European A. mellifera subspecies have been introduced ~60 years ago. Despite the high detection power and large sample size across Mongolian regions with beekeeping, the mite Acarapis woodi, the bacteria Melissococcus plutonius and Paenibacillus larvae, the microsporidian Nosema apis, Acute bee paralysis virus, Kashmir bee virus, Israeli acute paralysis virus and Lake Sinai virus strain 2 were not detected, suggesting that they are either very rare or absent. The mite Varroa destructor, Nosema ceranae and four viruses (Sacbrood virus, Black queen cell virus, Deformed wing virus (DWV) and Chronic bee paralysis virus) were found with different prevalence. Despite the positive correlation between the prevalence of V. destructor mites and DWV, some areas had only mites, but not DWV, which is most likely due to the exceptional isolation of apiaries (up to 600 km). Phylogenetic analyses of the detected viruses reveal their clustering and European origin, thereby supporting the role of trade for pathogen spread and the isolation of Mongolia from South-Asian countries. In conclusion, this survey reveals the distinctive honey bee pathosphere of Mongolia, which offers opportunities for exciting future research.

Tres nuevas citas de trips (Thysanoptera: Thripidae) para la Argentina y clave de los géneros de la familia Thripidae presentes en el país

Tres especies de trips se citan por primera vez para la Argentina. Scirtothrips inermis y Scolothrips pallidus se recolectaron sobre Viburnum tinus en la provincia de Mendoza. La primera de ellas es fitófaga y potencial plaga de citrus y la segunda es una especie benéfica y preda ácaros. También se registra por primera vez Thrips trehernei sobre Taraxacum officinale en Buenos Aires. Se elabora una clave para los géneros de la familia Thripidae en la Argentina. Asimismo, se presentan claves para las especies de los géneros Thrips y Scirtothrips en Argentina.

Fluctuación poblacional de dos eriófidos del olivo (Acari: Eriophyidae) en Coquimbito (Maipú, Mendoza, Argentina)

El cultivo del olivo actualmente es afectado por dos especies de eriófidos poco conocidas en Argentina. Estos organismos son fitófagos obligados de numerosas plantas, que infestan todos sus órganos, excepto las raíces; algunos causan daños económicamente significativos al provocar malformaciones en diferentes partes de la planta: falta de crecimiento, acortamiento de brotes, formación de escoba de brujas, torsión y decoloración de hojas. Las especies presentes en los olivares de Coquimbito, Maipú, son: Aceria oleae (Nalepa) y Oxycenus maxwelli (Keifer). Con el objetivo de establecer la fluctuación poblacional de ambas especies se realizó un monitoreo en un monte olivícola ubicado en Maipú, Mendoza. Se seleccionaron plantas con síntomas evidentes de la plaga. Se efectuó, quincenalmente, un muestreo dirigido, extrayendo 30 brotes y 30 inflorescencias o frutos con pedúnculo de los cuales se observaron, bajo estereomicroscopio, 100 hojas, 30 yemas vegetativas y 30 inflorescencias o frutos. Los resultados indican que estos ácaros pasan el invierno principalmente en las hojas y en las yemas vegetativas. A fines de septiembre se detecta una mayor proporción de individuos en yemas florales, flores y posteriormente en frutos recién cuajados, aumentando hasta llegar a su densidad máxima en diciembre.

Time series of zooplankton abundance at station L4 in the English Channel from 1988 to 2011

Ongoing zooplankton research at the Plymouth Marine Laboratory has established a time series of zooplankton species since 1988 at L4, a coastal station off Plymouth. Samples were collected by vertical net hauls (WP2 net, mesh 200 µm; UNESCO 1968) from the sea floor (approximately 50 m) to the surface and stored in 4% formalin. Much of the zooplankton analysis has been to the level of "major taxonomic groups" only, and a number of different analysts have participated over the years. The level of expertise has generally been consistent, but the user should be aware that levels of taxonomic discrimination may vary during the course of the dataset. The dominant calanoid copepods are generally well discriminated to species throughout. Calanus has not been routinely examined for species determination, the assumption being that the local population is entirely composed of Calanus helgolandicus. In certain years there has been a particular interest in Temora stylifera, Centropages cherchiae and other species reflected in the dataset. The lack of records in other previous years does not necessarily reflect species absence. We view it as essential for all users of L4 plankton data to establish and maintain contact with the nominated current data originators as well as fully consulting the metadata. While not impinging on free data access, this ensures that this large, species-rich but slightly complex species database is being used in the correct way, and any potential issues with the data are clarified. Furthermore, a proper dialogue with these local experts on the time series will enable where appropriate the most recent sampling timepoints to be used. The data can be downloaded from BODC or from doi:10.1594/PANGAEA.778092 as files for each year by searching for "L4 zooplankton". The most comprehensive dataset is the version downloadable directly from this page. The entire set of zooplankton samples is stored at the Plymouth Marine Laboratory in buffered formalin, and may be available for further taxonomic analysis on request.

Occurrence of the Oribatid Mite Trhypochthoniellus longisetus longisetus (Acari: Trhypochthoniidae) on Tilapia (Oreochromis niloticus).

Mites as parasites infesting fish have been described in a few case reports involving Histiostoma anguillarum, H. papillata, and Schwiebea estradai. We describe the unexpected occurrence of oribatid mites of the genus Trhypochthoniellus on farmed tilapia Oreochromis niloticus. The fish had mites on the skin, fins, and gills, as well as in the mouth. The morphological characteristics of the mites, observed by optical and scanning electron microscopy, were consistent with those described for T. longisetus longisetus. All stages of development were observed, suggesting that the mites were able to actively reproduce on fish

Dispersal of aphids, whiteflies and their natural enemies under photoselective nets

Integrated Pest Management of insects includes several control tactics, such as the use of photoselective nets, which may reduce the flight activity of insects. Limiting the dispersal of pests such as aphids and whiteflies is important because of their major role as vectors of plant viruses, while a minor impact on natural enemies is desired. In this study, we examined for the first time the dispersal ability of three vector species, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), Macrosiphum euphorbiae (Thomas) (Hemiptera: Aphididae) and Myzus persicae (Sulzer) (Hemiptera: Aphididae), in cages covered with photoselective nets. Contrary to the results obtained with aphids, the ability of the whitefly B. tabaci, to reach the target plant was reduced by photoselective nets. In a second set of experiments, the impact of UV-absorbing nets on the visual cues of two important predator species, Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) and Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae), was evaluated. The anthocorid was caught in higher numbers in traps placed under regular nets, whereas the mites preferably chose environments in which the UV radiation was attenuated. We have observed a wide range of effects that impedes generalization, although photoselective nets have a positive effect on pest management of whiteflies and aphids under protected environments.

Side effects of modern pesticides on adults of the predatory mite Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) under laboratory conditions

Amblyseius swirskii (Athias-Henriot) is a polyphagous predatory mite which feeds on pollen and small arthropod preys like whiteflies, thrips and mites. This species is widely used in IPM programs in greenhouses, being essential for its success, to obtain information about the non target effects of the pesticides currently used in those crops where the mite is artificially released. This work describes a laboratory contact residual test for evaluating lethal (mortality after 72 hour exposure to fresh residues) and sublethal effects (fecundity and fertility of the surviving mites) of eleven modern pesticides to adults of A. swirskii. Spiromesifen is lipogenesis inhibitor; flonicamid a selective feeding inhibitor with a mode of action not totally known; flubendiamide a modulator of the rhyanodin receptor, sulfoxaflor has a complex mode of action not totally ascertained; metaflumizone is a voltage dependent sodium channel blocker; methoxyfenozide is an IGR, spirotetramat inhibits lipids; abamectin and emamectin activate the Cl- channel; spinosad is a neurotix naturalyte and deltamethrin a pyrethroid used as positive standard. Selected pesticides are effective against different key pests present in horticultural crop areas and were always applied at the maximum field recommended concentration in Spain if registered, or at the concentration recommended by the supplier. Out of the tested pesticides, spiromesifen, flonicamid, flubendiamide, sulfoxaflor, metaflumizone, methoxyfenozide and spirotetramat were harmless to adults of the predatory mite (IOBC toxicity class 1). The rest of pesticides exhibited some negative effects: emamectin was slightly harmful (IOBC 2), deltamethrin moderately harmful (IOBC 3) and spinosad and abamectin harmful (IOBC 4). Further testing under more realistic conditions is needed for those pesticides having some harmful effect on the mite prior deciding their joint use or not. Key words: Amblyseius swirskii, adults, laboratory, residual test, spiromesifen, flonicamid, flubendiamide, sulfoxaflor, metaflumizone, methoxyfenozide, spirotetramat, emamectin, deltamethrin, abamectin, spinosad.

Effects of a bifenthrin-treated net on the natural enemies Aphidius colemani (Haliday) and Adalia bipunctata L. in a cucumber crop in Central Spain. Semi-field experiments

Pest management practices that rely on pesticides are growing increasingly less effective and environmentally inappropriate in many cases and the search of alternatives is under focus nowadays. Exclusion of pests from the crop by means of pesticide-treated screens can be an eco-friendly method to protect crops, especially if pests are vectors of important diseases. The mesh size of nets is crucial to determine if insects can eventually cross the barrier or exclude them because there is a great variation in insect size depending on the species. Long-lasting insecticide-treated (LLITN) nets, factory pre-treated, have been used since years to fight against mosquitoes vector of malaria and are able to retain their biological efficacy under field for 3 years. In agriculture, treated nets with different insecticides have shown efficacy in controlling some insects and mites, so they seem to be a good tool in helping to solve some pest problems. However, treated nets must be carefully evaluated because can diminish air flow, increase temperature and humidity and decrease light transmission, which may affect plant growth, pests and natural enemies. As biological control is considered a key factor in IPM nowadays, the potential negative effects of treated nets on natural enemies need to be studied carefully. In this work, the effects of a bifentrhin-treated net (3 g/Kg) (supplied by the company Intelligent Insect Control, IIC) on natural enemies of aphids were tested on a cucumber crop in Central Spain in autumn 2011. The crop was sown in 8x6.5 m tunnels divided in 2 sealed compartments with control or treated nets, which were simple yellow netting with 25 mesh (10 x 10 threads/cm2; 1 x 1 mm hole size). Pieces of 2 m high of the treated-net were placed along the lateral sides of one of the two tunnel compartments in each of the 3 available tunnels (replicates); the rest was covered by a commercial untreated net of a similar mesh. The pest, Aphis gossypii Glover (Aphidae), the parasitoid Aphidius colemani (Haliday) (Braconidae) and the predator Adalia bipunctata L. (Coccinellidae) were artificially introduced in the crop. Weekly sampling was done determining the presence or absence of the pest and the natural enemies (NE) in the 42 plants/compartment as well as the number of insects in 11 marked plants. Environmental conditions (temperature, relative humidity, UV and PAR radiation) were recorded. Results show that when aphids were artificially released inside the tunnels, neither its number/plant nor their distribution was affected by the treated net. A lack of negative effect of the insecticide-treated net on natural enemies was also observed. Adalia bipunctata did not establish in the crop and only a short term control of aphids was observed one week after release. On the other hand, A. colemani did establish in the crop and a more long-term effect on the numbers of aphids/plant was detected irrespective of the type of net. KEY WORDS: bifenthrin-treated net, Adalia bipunctata, Aphidius colemani, Aphis gossypii, semi-field

Occupational allergic multiorgan disease induced by wheat flour

Bakers are repeatedly exposed to wheat flour (WF) and may develop sensitization and occupational rhinoconjunctivitis and/or asthma to WF allergens.1 Several wheat proteins have been identified as causative allergens of occupational respiratory allergy in bakery workers.1 Testing of IgE reactivity in patients with different clinical profiles of wheat allergy (food allergy, wheat-dependent exercise-induced anaphylaxis, and baker's asthma) to salt-soluble and salt-insoluble protein fractions from WF revealed a high degree of heterogeneity in the recognized allergens. However, mainly salt-soluble proteins (albumins, globulins) seem to be associated with baker's asthma, and prolamins (gliadins, glutenins) with wheat-dependent exercise-induced anaphylaxis, whereas both protein fractions reacted to IgE from food-allergic patients.1 Notwithstanding, gliadins have also been incriminated as causative allergens in baker's asthma.2 We report on a 31-year-old woman who had been exposed to WF practically since birth because her family owned a bakery housed in the same home where they lived. She moved from this house when she was 25 years, but she continued working every day in the family bakery. In the last 8 years she had suffered from work-related nasal and ocular symptoms such as itching, watery eyes, sneezing, nasal stuffiness, and rhinorrhea. These symptoms markedly improved when away from work and worsened at work. In the last 5 years, she had also experienced dysphagia with frequent choking, especially when ingesting meats or cephalopods, which had partially improved with omeprazole therapy. Two years before referral to our clinic, she began to have dry cough and breathlessness, which she also attributed to her work environment. Upper and lower respiratory tract symptoms increased when sifting the WF and making the dough. The patient did not experience gastrointestinal symptoms with ingestion of cereal products. Skin prick test results were positive to grass (mean wheal, 6 mm), cypress (5 mm) and Russian thistle pollen (4 mm), WF (4 mm), and peach lipid transfer protein (6 mm) and were negative to rice flour, corn flour, profilin, mites, molds, and animal dander. Skin prick test with a homemade WF extract (10% wt/vol) was strongly positive (15 mm). Serologic tests yielded the following results: eosinophil cationic protein, 47 ?g/L; total serum IgE, 74 kU/L; specific IgE (ImmunoCAP; ThermoFisher, Uppsala, Sweden) to WF, 7.4 kU/L; barley flour, 1.24 kU/L; and corn, gluten, alpha-amylase, peach, and apple, less than 0.35 kU/L. Specific IgE binding to microarrayed purified WF allergens (WDAI-0.19, WDAI-0.53, WTAI-CM1, WTAI-CM2, WTAI-CM3, WTAI-CM16, WTAI-CM17, Tri a 14, profilin, ?-5-gliadin, Tri a Bd 36 and Tri a TLP, and gliadin and glutamine fractions) was assessed as described elsewhere.3 The patient's serum specifically recognized ?-5-gliadin and the gliadin fraction, and no IgE reactivity was observed to other wheat allergens. Spirometry revealed a forced vital capacity of 3.88 L (88%), an FEV1 of 3.04 L (87%), and FEV1/forced vital capacity of 83%. A methacholine inhalation test was performed following an abbreviated protocol,4 and the results were expressed as PD20 in cumulative dose (mg) of methacholine. Methacholine inhalation challenge test result was positive (0.24 mg cumulative dose) when she was working, and after a 3-month period away from work and with no visits to the bakery house, it gave a negative result. A chest x-ray was normal. Specific inhalation challenge test was carried out in the hospital laboratory by tipping WF from one tray to another for 15 minutes. Spirometry was performed at baseline and at 2, 5, 10, 15, 20, 30, 45, and 60 minutes after the challenge with WF. Peak expiratory flow was measured at baseline and then hourly over 24 hours (respecting sleeping time). A 12% fall in FEV1 was observed at 20 minutes and a 26% drop in peak expiratory flow at 9 hours after exposure to WF,

Three novel families of miniature inverted-repeat transposable elements are associated with genes of the yellow fever mosquito, Aedes aegypti

Three novel families of transposable elements, Wukong, Wujin, and Wuneng, are described in the yellow fever mosquito, Aedes aegypti. Their copy numbers range from 2,100 to 3,000 per haploid genome. There are high degrees of sequence similarity within each family, and many structural but not sequence similarities between families. The common structural characteristics include small size, no coding potential, terminal inverted repeats, potential to form a stable secondary structure, A+T richness, and putative 2- to 4-bp A+T-biased specific target sites. Evidence of previous mobility is presented for the Wukong elements. Elements of these three families are associated with 7 of 16 fully or partially sequenced Ae. aegypti genes. Characteristics of these mosquito elements indicate strong similarities to the miniature inverted-repeat transposable elements (MITEs) recently found to be associated with plant genes. MITE-like elements have also been reported in two species of Xenopus and in Homo sapiens. This characterization of multiple families of highly repetitive MITE-like elements in an invertebrate extends the range of these elements in eukaryotic genomes. A hypothesis is presented relating genome size and organization to the presence of highly reiterated MITE families. The association of MITE-like elements with Ae. aegypti genes shows the same bias toward noncoding regions as in plants. This association has potentially important implications for the evolution of gene regulation.