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Livestock production by smallholder farmers in southern Africa is often constrained by insufficient fodder during the dry season. The aim of this study was to determine if inclusion of pigeon pea (Cajanus cajan (L.) Millsp.) hedgerows in Megathyrsus maximus ‘Gatton’ pastures could improve fodder production and nutritional yields, and to investigate effects of hedgerow cutting height on fodder production (including both browse and herbage). Treatments included three simulated silvopastoral systems with M. maximus planted between hedgerows, and one treatment that was M. maximus pasture alone. Hedgerows were cut at 60 or 90 cm above ground, or were uncut. Over the first two growing seasons, browse and grass fodder was harvested, dried and weighed four times. A portion of the fodder (comprising browse and herbage) was conserved in the second year and samples analysed for crude protein (CP), neutral detergent fibre (NDF) and acid detergent fibre (ADF) contents (%) and CP, phosphorus (P) and zinc (Zn) yields (kg ha−1). Hedgerows decreased cumulative grass biomass by 18.5%, but increased 1.22-fold the cumulative total fodder biomass when hedgerows were cut to 90 cm, compared to no hedgerows. Hedgerows had no effect on conserved grass herbage biomass (2.7 Mg ha−1 on average), but doubled biomass of conserved total fodder (5.0 Mg ha−1 on average) if hedgerows were cut. Cutting hedgerows increased CP, P and Zn yields, and increased CP and ADF contents, compared to not cutting them or not including them, while NDF was unaffected. Cutting hedgerows to 90 cm increased total browse production, compared to cutting to 60 cm. Pigeon pea seed biomass was unaffected by hedgerow cutting height (870 kg ha−1 on average). In conclusion, pigeon pea hedgerows included into grass pastures increase the quantity and quality of total fodder for livestock feed supplementation during the dry season.

Inadequate nutrition limits small-scale livestock production by smallholder farmers in South Africa but can be addressed by the introduction of woody fodder species. Cutting woody plants to harvest fodder affects their productivity and thus it is necessary to determine the optimum cutting height. Cutting affects the productivity and survival of woody plants because it alters concentrations of non-structural carbohydrates, which are used to support their recovery. Since pigeon pea (Cajanus cajan (L.) Millsp.) is not widely grown in South Africa and is not managed as a pruned hedgerow, there was a need to investigate the effect of cutting height under local conditions, especially in areas that experience frost. The trial was established in November 2016, ran over two growing seasons (2016/2017 and 2017/2018) and concluded in August 2018. The treatments comprised three silvopastoral systems with Panicum maximum planted between pigeon pea hedgerows that were cut at 60 cm and 90 cm above the ground, or left uncut and sole P. maximum. Over the two year period, the pigeon pea plants cut at 90 cm above ground produced significantly more leaf, twig and total biomass than the plants cut at 60 cm. Cutting height also had a significant effect on the percentage twig, with pigeon pea plants cut at 90 cm having significantly higher percentage twig than those cut at 60 cm (P 0.05). In July 2018 there were a number of days with temperatures below 0 °C. Prior to this no mortalities had been observed, but the frost events resulted in die-off. The mortality rates were determined in August 2018 when it was clear which of the shrubs had clearly died. The shrub mortality rates were higher for pruned relative to unpruned shrubs, but the effect was not significant (P > 0.05). It was concluded that the choice of cutting height is important for maximising browse production. Future research should consider the effect of pruning on non-structural carbohydrate concentration and total pool in the roots to better understand the reasons for reduced production and shrub mortalities.

Some birds replace feathers after the formative plumage of the first cycle and after basic plumages in subsequent cycles as a consequence of prealternate molts. The presence of prealternate molts has been suggested to be due to prolonged exposure to sunlight, especially for birds inhabiting exposed and scrubby areas. Although prealternate molts appear to be common in many Nearctic-Neotropic passerines, these can be also present in resident Neotropical species. For example, prealternate molt has been reported in 14 species among nine genera in thraupids. Here, using wild individuals and museum specimens, we describe the presence of a Complex Alternate Strategy in the molt cycle of the Peru’s Blue-and-yellow Tanager (Pipraeidea bonariensis darwinii). We categorized seven molt cycle–based age groups in 75 wild-captured birds and thirteen molt cycle–based age categories across 167 museum specimens. Our results confirm the existence of limited to partial prealternate molts in Blue-and-yellow Tanager and demonstrate the importance of investigating the presence of these types of molts in wild bird populations.

A variety of genetic techniques have been devised to determine cell lineage relationships during tissue development. Some of these systems monitor cell lineages spatially and/or temporally without regard to gene expression by the cells, whereas others correlate gene expression with the lineage under study. The GAL4 Technique for Real-time and Clonal Expression (G-TRACE) system allows for rapid, fluorescent protein-based visualization of both current and past GAL4 expression patterns and is therefore amenable to genome-wide expression-based lineage screens. Here we describe the results from such a screen, performed by undergraduate students of the University of California, Los Angeles (UCLA) Undergraduate Research Consortium for Functional Genomics (URCFG) and high school summer scholars as part of a discovery-based education program. The results of the screen, which reveal novel expression-based lineage patterns within the brain, the imaginal disc epithelia, and the hematopoietic lymph gland, have been compiled into the G-TRACE Expression Database (GED), an online resource for use by the Drosophila research community. The impact of this discovery-based research experience on student learning gains was assessed independently and shown to be greater than that of similar programs conducted elsewhere. Furthermore, students participating in the URCFG showed considerably higher STEM retention rates than UCLA STEM students that did not participate in the URCFG, as well as STEM students nationwide.

Airborne pollen is a major cause of allergic rhinitis, affecting between 10 and 30% of the population in Belgium, the Netherlands, and Luxembourg (Benelux). Allergenic pollen is produced by wind pollinating plants and released in relatively low to massive amounts. Current climate changes, in combination with increasing urbanization, are likely to affect the presence of airborne allergenic pollen with respect to exposure intensity, timing as well as duration. Detailed analysis of long-term temporal trends at supranational scale may provide more comprehensive insight into these phenomena. To this end, the Spearman correlation was used to statistically compare the temporal trends in airborne pollen concentration monitored at the aerobiological stations which gathered the longest time-series (30–44 years) in the Benelux with a focus on the allergenic pollen taxa: Alnus, Corylus, Betula, Fraxinus, Quercus, Platanus, Poaceae, and Artemisia. Most arboreal species showed an overall trend toward an increase in the annual pollen integral and peak values and an overall trend toward an earlier start and end of the pollen season, which for Betula resulted in a significant decrease in season length. For the herbaceous species (Poaceae and Artemisia), the annual pollen integral and peak values showed a decreasing trend. The season timing of Poaceae showed a trend toward earlier starts and longer seasons in all locations. In all, these results show that temporal variations in pollen levels almost always follow a common trend in the Benelux, suggesting a similar force of climate change-driven factors, especially for Betula where a clear positive correlation was found between changes in temperature and pollen release over time. However, some trends were more local-specific indicating the influence of other environmental factors, e.g., the increasing urbanization in the surroundings of these monitoring locations. The dynamics in the observed trends can impact allergic patients by increasing the severity of symptoms, upsetting the habit of timing of the season, complicating diagnosis due to overlapping pollen seasons and the emergence of new symptoms due allergens that were weak at first.

Airborne pollen monitoring is of global socio-economic importance as it provides information on presence and prevalence of allergenic pollen in ambient air. Traditionally, this task has been performed by microscopic investigation, but novel techniques are being developed to automate this process. Among these, DNA metabarcoding has the highest potential of increasing the taxonomic resolution, but uncertainty exists about whether the results can be used to quantify pollen abundance. In this study, it is shown that DNA metabarcoding using trnL and nrITS2 provides highly improved taxonomic resolution for pollen from aerobiological samples from the Netherlands. A total of 168 species from 143 genera and 56 plant families were detected, while using a microscope only 23 genera and 22 plant families were identified. NrITS2 produced almost double the number of OTUs and a much higher percentage of identifications to species level (80.1%) than trnL (27.6%). Furthermore, regressing relative read abundances against the relative abundances of microscopically obtained pollen concentrations showed a better correlation for nrITS2 (R2 = 0.821) than for trnL (R2 = 0.620). Using three target taxa commonly encountered in early spring and fall in the Netherlands (Alnus sp., Cupressaceae/Taxaceae and Urticaceae) the nrITS2 results showed that all three taxa were dominated by one or two species (Alnus glutinosa/incana, Taxus baccata and Urtica dioica). Highly allergenic as well as artificial hybrid species were found using nrITS2 that could not be identified using trnL or microscopic investigation (Alnus × spaethii, Cupressus arizonica, Parietaria spp.). Furthermore, perMANOVA analysis indicated spatiotemporal patterns in airborne pollen trends that could be more clearly distinguished for all taxa using nrITS2 rather than trnL. All results indicate that nrITS2 should be the preferred marker of choice for molecular airborne pollen monitoring., Graphical abstract Unlabelled Image, Highlights • DNA successfully extracted from microscopic pollen slides and Burkard-collected tapes. • nrITS2 and trnL DNA metabarcoding improves taxonomic resolution of airborne pollen. • Relative read abundance nrITS2 shows higher correlation with pollen counts than trnL. • Finer scale spatiotemporal patterns in pollen trends detected using nrITS2. • Detection of artificial hybrid (Alnus × spaethii) that significantly prolongs hay fever season

Monitoring of airborne pollen concentrations provides an important source of information for the globally increasing number of hay fever patients. Airborne pollen is traditionally counted under the microscope, but with the latest developments in image recognition methods, automating this process has become feasible. A challenge that persists, however, is that many pollen grains cannot be distinguished beyond the genus or family level using a microscope. Here, we assess the use of Convolutional Neural Networks (CNNs) to increase taxonomic accuracy for airborne pollen. As a case study we use the nettle family (Urticaceae), which contains two main genera (Urtica and Parietaria) common in European landscapes which pollen cannot be separated by trained specialists. While pollen from Urtica species has very low allergenic relevance, pollen from several species of Parietaria is severely allergenic. We collect pollen from both fresh as well as from herbarium specimens and use these without the often used acetolysis step to train the CNN model. The models show that unacetolyzed Urticaceae pollen grains can be distinguished with > 98% accuracy. We then apply our model on before unseen Urticaceae pollen collected from aerobiological samples and show that the genera can be confidently distinguished, despite the more challenging input images that are often overlain by debris. Our method can also be applied to other pollen families in the future and will thus help to make allergenic pollen monitoring more specific.

Hay fever patients can be helped in managing their symptoms by predicting the severity of the upcoming grass pollen seasons., Allergic rhinitis is an inflammation in the nose caused by overreaction of the immune system to allergens in the air. Managing allergic rhinitis symptoms is challenging and requires timely intervention. The following are major questions often posed by those with allergic rhinitis: How should I prepare for the forthcoming season? How will the season’s severity develop over the years? No country yet provides clear guidance addressing these questions. We propose two previously unexplored approaches for forecasting the severity of the grass pollen season on the basis of statistical and mechanistic models. The results suggest annual severity is largely governed by preseasonal meteorological conditions. The mechanistic model suggests climate change will increase the season severity by up to 60%, in line with experimental chamber studies. These models can be used as forecasting tools for advising individuals with hay fever and health care professionals how to prepare for the grass pollen season.

Se estudió la dieta del suri Rhea pennata en la región de Moquegua al sur de Perú, mediante el análisis de heces. Esta especie considerada en “Peligro Crítico” en el Perú se mostró como estrictamente herbívora, principalmente folívora. Se identificaron 44 morfoespecies de plantas en su dieta, de las cuales Oxychloe andina (una especie de Juncaceae dominante en bofedales disturbados) presentó los mayores valores relativos de densidad, peso, volumen y frecuencia. A pesar de la aparente selectividad, esta especie presenta una dieta muy diversa que le permitiría adaptarse a las carencias nutricionales en el hábitat que ocupa. Los datos obtenidos en diferentes hábitats que ocupa el Suri, muestran que en zonas más áridas existe la tendencia a una mayor selectividad debido a la presencia de plantas con compuestos secundarios (fenoles, terpenos y alcaloides) para evitar la herbivoría, mientras que en ambientes con bofedales (como el del presente estudio) la selectividad disminuye, tendiendo a alimentarse de las especies dominantes., We study the Lesser Rhea (Rhea pennata) diet in the Moquegua region of southern Peru, using analysis of feces. This species considered “Critically Endangered” in Peru, was shown as strictly herbivorous, mainly folivorous. A total of 44 plant morphospecies were identified in the diet, of which Oxychloe andina (a Juncaceae species dominant in disturbed wetlands of the Puna) had the highest values of relative density, weight, volume and frequency. Despite the apparent selectivity, this species presents a very diverse diet that would allow it to adapt to the nutritional deficiencies in the habitat that it occupies. The data obtained in different habitats of the Lesser Rhea show that in more arid zones there is a tendency for greater selectivity due to the presence of plant with secondary compounds (phenols, terpenes and alkaloids) to avoid herbivory, while in environments with wetlands (as the present study) the selectivity decreases, tending to feed on the dominant species.

Changing from summer-brown to winter-white pelage or plumage is a crucial adaptation to seasonal snow in more than 20 mammal and bird species. Many of these species maintain nonwhite winter morphs, locally adapted to less snowy conditions, which may have evolved independently. Mountain hares ( Lepus timidus ) from Fennoscandia were introduced into the Faroe Islands in 1855. While they were initially winter-white, within ∼65 y all Faroese hares became winter-gray, a morph that occurs in the source population at low frequency. The documented population history makes this a valuable model for understanding the genetic basis and evolution of the seasonal trait polymorphism. Through whole-genome scans of differentiation and single-nucleotide polymorphism (SNP) genotyping, we associated winter coat color polymorphism to the genomic region of the pigmentation gene Agouti , previously linked to introgression-driven winter coat color variation in the snowshoe hare ( Lepus americanus ). Lower Agouti expression in the skin of winter-gray individuals during the autumn molt suggests that regulatory changes may underlie the color polymorphism. Variation in the associated genomic region shows signatures of a selective sweep in the Faroese population, suggesting that positive selection drove the fixation of the variant after the introduction. Whole-genome analyses of several hare species revealed that the winter-gray variant originated through introgression from a noncolor changing species, in keeping with the history of ancient hybridization between the species. Our findings show the recurrent role of introgression in generating winter coat color variation by repeatedly recruiting the regulatory region of Agouti to modulate seasonal coat color change.

Network metrics are widely used to infer the roles of mutualistic animals in plant communities and to predict the effect of species’ loss. However, their empirical validation is scarce. Here we parameterized a joint species model of frugivory and seed dispersal with bird movement and foraging data from tropical and temperate communities. With this model, we investigate the effect of frugivore loss on seed rain, and compare our predictions to those of standard coextinction models and network metrics. Topological coextinction models underestimated species loss after the removal of highly linked frugivores with unique foraging behaviours. Network metrics informed about changes in seed rain quantity after frugivore loss. However, changes in seed rain composition were only predicted by partner diversity. Nestedness, closeness, and d’ specialisation could not anticipate the effects of rearrangements in plant–frugivore communities following species loss. Accounting for behavioural differences among mutualists is critical to improve predictions from network models. Fil: Morán López, Teresa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Espíndola, Walter. State University of Pennsylvania; Estados Unidos Fil: Vizzachero, Benjamin. State University of Pennsylvania; Estados Unidos Fil: Fontanella, Antonio. Universidad Nacional Mayor de San Marcos; Perú Fil: Salinas, Letty. Universidad Nacional Mayor de San Marcos; Perú Fil: Arana, César. Universidad Nacional Mayor de San Marcos; Perú Fil: Amico, Guillermo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Pizo, Marco A.. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil Fil: Carlo, Tomás A.. Universidad Nacional Mayor de San Marcos; Perú. State University of Pennsylvania; Estados Unidos Fil: Morales, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina

Summary The clinical importance of anterior foregut endoderm (AFE) derivatives, such as thyrocytes, has led to intense research efforts for their derivation through directed differentiation of pluripotent stem cells (PSCs). Here, we identify transient overexpression of the transcription factor (TF) NKX2-1 as a powerful inductive signal for the robust derivation of thyrocyte-like cells from mouse PSC-derived AFE. This effect is highly developmental stage specific and dependent on FOXA2 expression levels and precise modulation of BMP and FGF signaling. The majority of the resulting cells express thyroid TFs (Nkx2-1, Pax8, Foxe1, Hhex) and thyroid hormone synthesis-related genes (Tg, Tpo, Nis, Iyd) at levels similar to adult mouse thyroid and give rise to functional follicle-like epithelial structures in Matrigel culture. Our findings demonstrate that NKX2-1 overexpression converts AFE to thyroid epithelium in a developmental time-sensitive manner and suggest a general methodology for manipulation of cell-fate decisions of developmental intermediates., Graphical Abstract, Highlights • Transient Nkx2-1 overexpression robustly converts foregut endoderm to thyroid lineage • The effect is highly developmental stage specific and dependent on BMP/FGF signals • The progeny self-organizes to 3D structures reminiscent of follicular thyroid tissue • Mature cultures produce T4 hormone at levels comparable with mouse thyroid tissue, Ikonomou and colleagues found that pluripotent stem cell-derived anterior foregut endoderm can be robustly converted to thyroid progenitors through transient expression of the transcription factor Nkx2-1. This inductive effect is highly stage specific and relies on thyroid specification signals. The thyrocyte-like progeny self-organize into follicle structures in 3D culture and express mature thyroid proteins indicative of functionality (TG, NIS, T4).

In contrast to the exploration and use of southern African grass plant genetic resources over the past century, only a few indigenous legume species are currently used as forages, notwithstanding the vast untapped legume diversity available in southern Africa. The aim of this study was to assess legumes, indigenous to South Africa, Lesotho and Swaziland and propose a list of species with pasture potential for further evaluation. Towards this end, legume species documented as being cultivated, grazed or browsed as well as plant collection data from the National Herbarium, South Africa, were used. The majority of cultivated, grazed or browsed species were recorded in the Central Bushveld, Lowveld and Mopane Bioregions, the Drakensberg Foothill and Coastal Region, Savanna Group and Northern Mistbelt leguminochoria and is largely enclosed by the Wolkberg Centre of Plant Endemism. The Phaseoleae tribe was found to contain the highest percentage of cultivated legume species. Legume species adapted to low soil phosphorus levels were identified, with 22 of these species previously noted for their pasture potential. It is suggested that the results of this study be used in the continued search for alternative indigenous legumes species for eventual integration into local and international pasture systems.Keywords: Fabaceae, fodder crops, Leguminosae, phosphorus, plant genetic resources

Cystic echinococcosis (CE) is an important helminthic zoonotic disease that commonly affects the liver and lungs. Imaging methods and serology establish the diagnosis in most cases. Chest x-ray can diagnose uncomplicated pulmonary hydatid cysts, whereas superinfection and/or rupture of the hydatid cyst (complicated cysts) may change the radiographic appearance and lead to delayed diagnosis and treatment. We report the case of a patient with hemoptysis and chest pain, where computer tomography scan of the lung suggested a large, ruptured hydatid cyst. However, serological tests with indirect hemagglutination (IHA)for Echinococcus granulosus antibodies were negative, and there was massive growth of Streptococcus pneumoniae in sputum. Based on this, we concluded that the patient had a bacterial lung abscess. The diagnosis of CE was only made after surgical removal of the cyst followed by microscopy and polymerase chain reaction.

Although RNA interference (RNAi) has become a ubiquitous laboratory tool since its discovery 12 years ago, in vivo delivery to selected cell types remains a major technical challenge. Here, we report the use of lentiviral vectors for long-term in vivo delivery of RNAi selectively to resident alveolar macrophages (AMs), key immune effector cells in the lung. We demonstrate the therapeutic potential of this approach by RNAi-based downregulation of p65 (RelA), a component of the pro-inflammatory transcriptional regulator, nuclear factor κB (NF-κB) and a key participant in lung disease pathogenesis. In vivo RNAi delivery results in decreased induction of NF-κB and downstream neutrophilic chemokines in transduced AMs as well as attenuated lung neutrophilia following stimulation with lipopolysaccharide (LPS). Through concurrent delivery of a novel lentiviral reporter vector (lenti-NF-κB-luc-GFP) we track in vivo expression of NF-κB target genes in real time, a critical step towards extending RNAi-based therapy to longstanding lung diseases. Application of this system reveals that resident AMs persist in the airspaces of mice following the resolution of LPS-induced inflammation, thus allowing these localized cells to be used as effective vehicles for prolonged RNAi delivery in disease settings.

SummaryTwo populations of Nkx2-1+ progenitors in the developing foregut endoderm give rise to the entire postnatal lung and thyroid epithelium, but little is known about these cells because they are difficult to isolate in a pure form. We demonstrate here the purification and directed differentiation of primordial lung and thyroid progenitors derived from mouse embryonic stem cells (ESCs). Inhibition of TGFβ and BMP signaling, followed by combinatorial stimulation of BMP and FGF signaling, can specify these cells efficiently from definitive endodermal precursors. When derived using Nkx2-1GFP knockin reporter ESCs, these progenitors can be purified for expansion in culture and have a transcriptome that overlaps with developing lung epithelium. Upon induction, they can express a broad repertoire of markers indicative of lung and thyroid lineages and can recellularize a 3D lung tissue scaffold. Thus, we have derived a pure population of progenitors able to recapitulate the developmental milestones of lung/thyroid development.

The diversity of legumes, indigenous to South Africa, Lesotho and Swaziland is reported using recorded descriptive and distribution data. A total of 24 tribes, 118 genera and 1662 species were documented with the majority of genera belonging to subfamily Faboideae, tribe Phaseoleae, and the majority of species to tribe Crotalarieae. In terms of distribution patterns, most species were present in the Savanna Biome and Central Bushveld Bioregion. Even though most tribes contain species with secondary metabolites (mainly non-protein amino acids), tribes containing poisonous species are far fewer. Herbs are the key growth form, followed by shrubs, dwarf shrubs and trees. The majority of species are perennials. A map representing the collection intensity for the study area showed that the majority of legumes species were collected in the Fynbos, Savanna and Grassland Biome. It is concluded that indigenous South African legumes are extremely diverse and this denotes the importance of further investigating their forage potential or use in other agricultural practices. Key words : Biomes, bioregions, Fabaceae, fodder, genebank, Leguminosae, pastures, tribes

Summary Coxiella burnetii, a Gram-negative obligate intracellular pathogen, replicates within an parasitophorous vacuole with lysosomal characteristics. To understand how C. burnetii maintains genomic integrity in this environment, a database search for genes involved in DNA repair was performed. Major components of repair, SOS response and recombination were identified, including recA and ruvABC, but lexA and recBCD were absent. Instead, C. burnetii possesses addAB orthologous genes, functional equivalents to recBCD. Survival after treatment with UV, mitomycin C (MC) or methyl methanesulfonate (MMS), as well as homologous recombination in Hfr mating was restored in Escherichia coli deletion strains by C. burnetii recA or addAB. Despite the absence of LexA, co-protease activity for C. burnetii RecA was demonstrated. Dominant-negative inhibition of C. burnetii RecA by recA mutant alleles, modelled after E. coli recA1 and recA56, was observed and more apparent with expression of C. burnetii RecAG159D mutant protein. Expression of a subset of repair genes in C. burnetii was monitored and, in contrast to the non-inducible E. coli recBCD, addAB expression was strongly upregulated under oxidative stress. Constitutive SOS gene expression due to the lack of LexA and induction of AddAB likely reflect a unique repair adaptation of C. burnetii to its hostile niche.

Inherited mutations in the human alpha(1)-antitrypsin (AAT) gene lead to deficient circulating levels of AAT protein and a predisposition to developing emphysema. Gene therapy for individuals deficient in AAT is an attractive goal, because transfer of a normal AAT gene into any cell type able to secrete AAT should reverse deficient AAT levels and attenuate progression of lung disease. Here we present an approach for AAT gene transfer based on the transplantation of lentivirally transduced hematopoietic stem cells (HSCs). We develop a novel dual-promoter lentiviral system to transfer normal human AAT cDNA as well as a fluorescent tracking "reporter gene" into murine HSCs. After transplantation of 3,000 transduced HSCs into irradiated mouse recipients, we demonstrate simultaneous and sustained systemic expression of both genes in vivo for at least 31 weeks. The stem cells transduced with this protocol maintain multipotency, self-renewal potential, and the ability to reconstitute the hematopoietic systems of both primary and secondary recipients. This lentiviral-based system may be useful for investigations requiring the systemic secretion of anti-proteases or cytokines relevant to the pathogenesis of a variety of lung diseases.

The present study determines lead content in onion root tip cells (Allium cepa L.), correlating it with its toxicity. The treatment was carried at 25 +/- 0.5 degrees C using aqueous solutions of lead chloride at 0.1, 0.25, 0.50, 0.75, and 1 ppm for 12, 24, 48, and 72 h. For each treatment, a control where the lead solution was substituted by distilled water was included. After treatment, the meristems were fixed with a mixture of alcohol-acetic acid (3:1) and colored according to the technique of Feulgen. Lead content was quantified by graphite furnace absorption atomic spectrometry. The lead content in the roots ranged from 3.25 to 244.72 microg/g dry weight, with a direct relation with the concentration and time of exposure. A significant negative correlation was presented (r = -0.3629; p < 0.01) among lead content and root growth increment, and a positive correlation (r = 0.7750; p < 0.01) with the induction of chromosomic aberrations. In conclusion, lead is able to induce a toxic effect in the exposed roots, correlated with its content.

The translocation of animal species, that is the transfer of individuals from a source population to a release area to restock, is a widespread practice in conservation biology and wildlife management. However, translocation is likely a severe test for the translocated individuals as they are subjected to stressful and inevitable disturbances: physical handling, transport and release into an unknown environment. Moreover, other disturbances may be induced by translocation, for example in case of social disorganization among translocated individuals or of a large difference in habitat quality between origin and release areas. These disturbances induced by the translocation process may have a negative impact upon the translocated individuals, through reduced survival or breeding success, or abnormal behaviour. Consequently, founding a viable population by the mean of a translocation is not easy. Then, it is necessary to determine which factors are involved in the failure or the success of a translo...

En la región altoandina de Ancash la mayor diversidad de flores ornitófilas se encuentran en ambientes de matorral, en contraste a roquedales y pajonales, en los que encontramos rodales de bromelias del género Puya, cuyo néctar podría constituir un importante recurso para picaflores altoandinos en estos tipos de ambiente. Para documentar esta hipótesis, entre el 2004 y el 2005 se realizaron un total de 264 horas de observación de picaflores de dos rodales de Puya, ubicados encima de los 3000 m de altitud. El primer rodal fue de Puya raimondii en un pajonal del Parque Nacional Huascarán (9º39' S-77º13' W), el segundo fue de Puya rauhii en roquedales del Callejón de Conchucos (8º10' S-77º52' W). La frecuencia de visita de los picaflores a las inflorescencias de Puya fue evaluada desde 10 puntos fijos de observación, además capturas con redes de niebla permitieron identificar el polen de su pico y frente. En P. raimondii se identificaron cuatro especies de Trochilidae alimentándose de su néctar (Oreotrochilus stolzmanni, Patagona gigas, Aglaeactis cupripennis y Metallura phoebe), siendo O. stolzmanni la especie más frecuentemente avistada (60%). El 80% de las muestras de polen obtenidas de estos picaflores correspondían a P. raimondii. En el rodal de Puya rauhii se identificaron diez especies de Trochilidae (Colibri coruscans, Oreotrochilus stolzmanni, Patagona gigas, Aglaeactis cupripennis, Coeligena iris, Chalcostigma stanleyi, Lesbia nuna, Myrtis fanny, Metallura phoebe y Metallura tyrianthina), siendo las especies con mayores avistamientos M. phoebe (26%), C. coruscans (21%) y P. gigas (17%). El 31% de las muestras de polen obtenidas de estos picaflores correspondían a Puya. Las especies altoandinas de Puya proporcionan un importante recurso alimenticio para picaflores, en especial para los de grandes altitudes donde la diversidad de plantas disminuye. In the high Andean region of Ancash the greatest diversity of flowers ornithophilous is in shrublands, in contrast to rocky areas and grasslands, in those that we find stands of bromelias of the genera Puya whose nectar could constitute an important resource for high Andean hummingbirds in these habitat types. To document this hypothesis, during 2004-2005, in 264 hours of observation, were evaluated the hummingbirds of two stands of Puya above the 3000 m of altitude. The first stand was of Puya raimondii in a grassland of the Huascarán National Park (9º39' S-77º13' W), the second was of Puya rauhii in rocky areas of the Conchucos Valley (8º10' S-77º52' W). The frequency of visit of hummingbirds to the inflorescences of Puya was evaluated from 10 fixed points of observation, also captures with mist nets allowed to identify the pollen of their bill and front. In P. raimondii four species of Trochilidae were identified feeding of their nectar (Oreotrochilus stolzmanni, Patagonia gigas, Aglaeactis cupripennis and Metallura phoebe), being O. stolzmanni the frequently sighted species (60%). 80% of the obtained samples of pollen of these hummingbirds corresponded P. raimondii. In the rodal of Puya rauhii was identified ten species of Trochilidae (Colibri coruscans, Oreotrochilus stolzmanni, Patagonia gigas, Aglaeactis cupripennis, Coeligena iris, Chalcostigma stanleyi, Lesbia nuna, Myrtis fanny, Metallura phoebe and M. tyrianthina), being the species with more sightings M. phoebe (26%), C. coruscans (21%) and P. gigas (17%). 31% of the obtained samples of pollen of these hummingbirds corresponded to Puya. The high Andean species of Puya provide an important nutritious resource for hummingbirds, especially for those of high altitudes where the diversity of plants diminishes.

Quantification of airborne pollen is an important tool in scientific research and patient care in allergy. The currently available method relies on microscopic examination of pollen slides, performed by qualified researchers. Although highly reliable, the method is labor intensive and requires extensive training of the researchers involved. In an approach to develop alternative detection methods, we performed a feasibility study on the automated recognition of the allergenic relevant pollen, grass, birch, and mugwort, by utilizing digital image analysis and pattern recognition tools. Of a total of 254 pollen samples (including 79 of grass, 79 of birch and 96 of mugwort), 97.2% were recognized correctly. This encouraging result provides a promising prospect for future developments.

The vanadium is a metal that presents great interest from the toxicological point of view, because of the numerous alterations that can take place in different biological systems. This work evaluated the capacity of vanadium accumulation and its correlation with genotoxic effects in root cells of Allium cepa L. The bulbs were cultivated in renovated filtered water each 24 h, at a temperature of 25 +/- 0.5 degrees C, in darkness and constant aeration. Treatments were carried out under the same experimental conditions, using water solutions of vanadium of 25, 50, 75 and 100 microg/g for 0, 12, 24, 48 and 72 h. A control was carried out where metal solution was substituted by distilled water. After the treatment, the meristems were fixed with alcohol--acetic acid (3:1) and stained according to the technique of Feulgen. The capacity of accumulation was determined by GFAAS. The analysis of the results revealed an accumulation of the metal for all times and concentrations. No correlation was presented among vanadium accumulation, growth and mitotic index; however, positive correlation was given with the induction of chromosomic aberrations. In conclusion, vanadium is able to induce cytotoxic effect in the exposed roots, but only genotoxic effect was correlated with metal accumulation.

Restocking is a widespread management practice used to support wild populations of European rabbit Oryctolagus cuniculus which are jeopardised by habitat loss and disease outbreaks. However, rabbits are known to experience a high mortality just after release, which might be due to the handling stress induced by translocation, and particularly to storage condition between capture and release. In our study, we tested two soft treatments of storage in boxes, where rabbits were stored either alone or in groups of four or five, and we assessed both survival rates and maximum dispersal distances by radio-tracking 43 individuals during eight weeks after release. We also recorded data about the survival at the time of release for 11 other rabbits that either died before release or were not radio-equipped. Our study involved two similar experimental releases in 2001 and 2002 in distinct nearby study areas to assess the effect of storage condition. Rabbits were released in two subsequent batches at a few d...

8 p.-5 fig.-3 tab., This work evaluated the qualitative and quantitative cellular changes induced by treatment with 5-aminouracil (5-AU) and a combination of 5-AU and caffeine in plant cells in relation to DNA damage, repaired damage, and residual damage. As biological material, Allium cepa L. root tips were used, grown in filtered water, in darkness, with aeration at constant temperature of 25 °C ± 0.5. Cell populations were synchronized using 5 mM caffeine in order to study the effects of 5-AU and caffeine/5-AU combined treatment on the DNA content and their incidence in the entrance to mitosis. The results showed a delay in the G2 period due to induced DNA damage by the 5-AU and caffeine/5-AU combined treatment, shown by aberrant metaphases, anaphases and telophases. The effect of caffeine in the combined treatment was heightened in spite of lengthening the checkpoints route that retains the cells in G2. The existence of G2 checkpoints was shown in the cell population studied, inducing lesions in the DNA, chromosomic aberrations and cellular instability., This work was financed by the Fondo Nacional para la Ciencia y la Tecnología de Venezuela (FONACIT).

Serological data on myxoma virus, rabbit haemorrhagic disease (RHD) virus and RHD-like viruses in juvenile rabbits (Oryctolagus cuniculus) trapped in 1995, 1996 and 1997 in two areas of France were analysed. For each disease, the effects of bodyweight, year, month and seropositivity for the other disease were modelled by using logistic regressions. In one area, a model including RHD seropositivity was selected to explain the myxoma virus seropositivity. Models including myxoma virus seropositivity were selected to explain the RHD seropositivity in both areas, and the odds of a rabbit being seropositive to both viruses were 5.1 and 8.4 times higher than the odds of a rabbit being seronegative to myxoma virus and seropositive to RHD. The year and bodyweight had significant effects for myxomatosis in one area and for RHD in both areas.

Glucagon has a short plasma t1/2in vivo, with renal extraction playing a major role in its elimination. Glucagon is degraded by neutral endopeptidase (NEP) 24.11 in vitro, but the physiological relevance of NEP 24.11 in glucagon metabolism is unknown. Therefore, the influence of candoxatril, a selective NEP inhibitor, on plasma levels of endogenous and exogenous glucagon was examined in anesthetized pigs. Candoxatril increased endogenous glucagon concentrations, from 6.3 ± 2.5 to 20.7 ± 6.3 pmol/l [COOH-terminal (C)-RIA, P < 0.05]. During glucagon infusion, candoxatril increased the t1/2determined by C-RIA (from 3.0 ± 0.5 to 17.0 ± 2.5 min, P < 0.005) and midregion (M)-RIA (2.8 ± 0.5 to 17.0 ± 3.0 min, P < 0.01) and reduced metabolic clearance rates (MCR; 19.1 ± 3.2 to 9.4 ± 2.0 ml·kg−1·min−1, P < 0.02, C-RIA; 19.2 ± 4.8 to 9.0 ± 2.3 ml·kg−1·min−1, P < 0.05, M-RIA). However, neither t1/2nor MCR determined by NH2-terminal (N)-RIA were significantly affected ( t1/2, 2.7 ± 0.4 to 4.5 ± 1.6 min; MCR, 30.3 ± 6.4 to 28.5 ± 9.0 ml·kg−1·min−1), suggesting that candoxatril had no effect on NH2-terminal degradation but leads to the accumulation of NH2-terminally truncated forms of glucagon. Determination of arteriovenous glucagon concentration differences revealed that renal glucagon extraction was reduced (but not eliminated) by candoxatril (from 40.4 ± 3.8 to 18.6 ± 4.1%, P < 0.02, C-RIA; 29.2 ± 3.1 to 14.7 ± 2.2%, P < 0.02, M-RIA; 26.5 ± 4.0 to 19.7 ± 3.5%, P < 0.06, N-RIA). Femoral extraction was reduced by candoxatril when determined by C-RIA (from 22.7 ± 2.4 to 8.0 ± 5.1%, P < 0.05) but was not changed significantly when determined using M- or N-RIAs (10.0 ± 2.8 to 4.7 ± 3.7%, M-RIA; 10.5 ± 2.5 to 7.8 ± 4.2%, N-RIA). This study provides evidence that NEP 24.11 is an important mediator of the degradation of both endogenous and exogenous glucagon in vivo.

Maleic hydrazide (MH) is an herbicide and is a regulator of the growth of buds in vegetables during storage. It is used in agriculture-in despite its known effect as a mutagenic and clastogenic agent. In this research the effect of MH on the root tips of Allium cepa L. was determined; correlations between the effects of different concentrations and exposure times on the mitotic index (MI) and induction of chromosomal aberrations (ChA) were also examined. Experiments were carried out in triplicate, using aqueous solutions of MH to concentrations of 10(-6), 10(-5), 10(-4) and 10(-3)M, at intervals of 0, 4, 8, 12, 24, and 48 h, with a control for each combination (with the MH substituted by distilled water). The results revealed an inhibition of the MI linked to the concentration and time of treatment (F=845.51, P

The polymorphic human debrisoquine hydroxylase, cytochrome P450 2D6 (CYP2D6), is one of the most important phase I drug metabolising enzymes. It is responsible for metabolising a large number of compounds that mostly share similarity in having a basic N-atom and an aromatic moiety. In homology modelling studies, it has been suggested that in fixation of this aromatic moiety, there may be an important role for phenylalanine 120 (Phe(120)). In this study, the role of Phe(120) in ligand binding and catalysis was experimentally examined by mutating it into an alanine. Strikingly, this substitution led to a completely abolished 7-methoxy-4-(aminomethyl)-coumarin (MAMC) O-demethylating activity of CYP2D6. On the other hand, bufuralol metabolism was hardly affected (K(m) of 1-hydroxylation mutant: 1.2 microM, wild-type: 2.9 microM, 4-hydroxylation mutant: 1.5 microM, and wild-type: 3.2 microM) and neither was affected dextromethorphan O-demethylation (K(m) mutant: 1.2 microM, wild-type: 2 microM, k(cat) mutant: 4.5 min(-1), and wild-type: 3.3 min(-1)). However, the Phe(120)Ala mutant also formed 3-hydroxymorphinan, the double demethylated form of dextromethorphan, which was not detected using wild-type CYP2D6. 3,4-Methylenedioxymethamphetamine (MDMA) was demethylenated by both mutant and wild-type CYP2D6 to 3,4-dihydroxymethamphetamine (3,4-OH-MA K(m) of mutant: 55 microM and wild-type: 2 microM). In addition, the mutant formed two additional metabolites; 3,4-methylenedioxyamphetamine (MDA) and N-hydroxy-3,4-methylenedioxymethamphetamine (N-OH-MDMA). Inhibition experiments of dextromethorphan O-demethylation showed a decreased affinity of the Phe(120)Ala mutant for quinidine (IC(50) mutant: 240 nM and wild-type, 40 nM), while IC(50)s for quinine were equal (1 microM). These data indicate the importance of Phe(120) in the selectivity and regiospecificity in substrate binding and catalysis by CYP2D6.

Glucagon metabolism under basal (endogenous) conditions and during intravenous glucagon infusion was studied in anesthetized pigs by use of midregion (M), COOH-terminal (C), and NH2-terminal (N)-RIAs. Arteriovenous concentration differences revealed a negative extraction of endogenous glucagon immunoreactivity across the portal bed (-35.4 ± 11.0, -40.3 ± 9.6, -35.6 ± 16.9%, M-, C-, N-RIA, respectively), reflecting net secretion of pancreatic glucagon and intestinal glicentin and oxyntomodulin, but under exogenous conditions, a net extraction occurred (11.6 ± 3.6 and 18.6 ± 5.7%, C- and N-RIA, respectively). Hindlimb extraction of endogenous (17.4 ± 3.7%, C-RIA) and exogenous (29.1 ± 4.8 and 19.8 ± 5.1%, C- and M-RIA) glucagon was detected, indicating M and C cleavage of the molecule. Renal extraction of glucagon was detected by all assays under endogenous (19.4 ± 6.7, 33.9 ± 7.1, 29.5 ± 6.7%, M-, C-, N-RIA) and exogenous conditions (46.9 ± 4.8, 46.4 ± 6.0, 47.0 ± 7.7%; M-, C-, N-RIA), indicating substantial elimination of the peptide. Hepatic glucagon extraction was undetectable under basal conditions and detected only by M-RIA (10.0 ± 3.8%) during glucagon infusion, indicating limited midregional cleavage of the molecule. The plasma half-life determined by C- and N-RIAs (2.7 ± 0.2 and 2.3 ± 0.2 min) were similar, but both were shorter than when determined by M-RIA (3.2 ± 0.2 min, P < 0.02). Metabolic clearance rates were similar regardless of assay (14.4 ± 1.1, 13.6 ± 1.7, 17.0 ± 1.7 ml·kg-1·min-1, M-, C-, N-RIA). Porcine plasma degraded glucagon, but this was not significantly affected by the dipeptidyl peptidase IV (DPP IV) inhibitor valine-pyrrolidide, and in anesthetized pigs, glucagon's metabolic stability was unchanged by DPP IV inhibition. We conclude that tissue-specific metabolism of glucagon occurs, with the kidney being the main site of removal and the liver playing little, if any, role. Furthermore, valine-pyrrolidide has no effect on glucagon stability, suggesting that DPP IV is unimportant in glucagon metabolism in vivo, in contrast to its significant role in the metabolism of the other proglucagon-derived peptides and glucose-dependent insulinotropic polypeptide.

International audience; no abstract

Wild rabbit (Oryctolagus cuniculus) restocking is a common practice in management in France. However rabbits generally experience heavy mortality after release, which jeopardizes restocking efficiency. Release success is full only when introduced individuals produce offspring.

One-third of the Dutch population suffers from allergic rhinitis, including hay fever. In this study, a 5-day-ahead hay fever forecast was developed and validated for grass pollen allergic patients in the Netherlands. Using multiple regression analysis, a two-step pollen and hay fever symptom prediction model was developed using actual and forecasted weather parameters, grass pollen data and patient symptom diaries. Therefore, 80 patients with a grass pollen allergy rated the severity of their hay fever symptoms during the grass pollen season in 2007 and 2008. First, a grass pollen forecast model was developed using the following predictors: (1) daily means of grass pollen counts of the previous 10 years; (2) grass pollen counts of the previous 2-week period of the current year; and (3) maximum, minimum and mean temperature (R 2 = 0.76). The second modeling step concerned the forecasting of hay fever symptom severity and included the following predictors: (1) forecasted grass pollen counts; (2) day number of the year; (3) moving average of the grass pollen counts of the previous 2 week-periods; and (4) maximum and mean temperatures (R 2 = 0.81). Since the daily hay fever forecast is reported in three categories (low-, medium- and high symptom risk), we assessed the agreement between the observed and the 1- to 5-day-ahead predicted risk categories by kappa, which ranged from 65 % to 77 %. These results indicate that a model based on forecasted temperature and grass pollen counts performs well in predicting symptoms of hay fever up to 5 days ahead.

Understanding relations between climate and pollen production is important for several societal and ecological challenges, importantly pollen forecasting for pollinosis treatment, forensic studies, global change biology, and high-resolution palaeoecological studies of past vegetation and climate fluctuations. For these purposes, we investigate the role of climate variables on annual-scale variations in pollen influx, test the regional consistency of observed patterns, and evaluate the potential to reconstruct high-frequency signals from sediment archives. A 43-year pollen-trap record from the Netherlands is used to investigate relations between annual pollen influx, climate variables (monthly and seasonal temperature and precipitation values), and the North Atlantic Oscillation climate index. Spearman rank correlation analysis shows that specifically in Alnus, Betula, Corylus, Fraxinus, Quercus and Plantago both temperature in the year prior to (T-1), as well as in the growing season (T), are highly significant factors (TApril rs between 0.30 [P

The Viral Activation Transfusion Study (VATS) was a randomized trial that compared leukocyte-reduced transfusions with unfiltered red blood cell transfusions in HIV and cytomegalovirus (CMV) antibody-positive patients with anemia who were undergoing their first blood transfusion. The relations of the baseline qualitative and quantitative polymerase chain reaction (PCR) measures of plasma CMV viremia, HIV RNA, CD4 + cell counts, and quality of life in these study subjects were examined. The 511 study subjects had a median CD4 + cell count equal to 15 cells/mm 3 , and 110 (21.5%) had CMV viremia by qualitative assay. In multivariate models, frequency of positive qualitative CMV increased with decreasing CD4 + cell counts (p =.04 trend), higher HIV RNA (p

Translocation is an important tool in conservation biology. However, translocation success is generally low for numerous animal species, therefore experiments are required for improvement. We carried out an experimental translocation of European wild rabbits (Oryctolagus cuniculus). The results may be of great interest for conservation biology, although we used a common species. As rabbits are known to experience a high mortality during the first days following release, it was necessary to assess the influence of handling trauma and environment novelty, respectively, and ways of suppressing them. Both tranquillization treatment during handling and a ‘soft’ release protocol (acclimatization pens in the new territory) were tested. Tranquillization did not increase survival, while the effect of acclimatization depended on sex. Females survived better when acclimatized, while males showed the opposite tendency. This difference is discussed in terms of sex-specific social behaviour, which is possibly an important correlate of translocation success. Finally, environmental stress seemed to override handling stress in determining the level of early survival for translocated wild rabbits.

Background Movements of animals have important consequences, at both the individual and population levels. Due to its important implications in the evolutionary dynamics of populations, dispersal is one of the most studied types of movement. In contrast, non-permanent extra home-range movements are often paid less attention. However, these movements may occur in response to important biological processes such as mating or predation avoidance. In addition, these forays are often preludes to permanent dispersal, because they may help individuals gain cues about their surroundings prior to settlement in a new place. In the European hare, exploration forays occur predominantly in juveniles, the time at which most hares disperse. In France, the timing of dispersal also overlaps with the hare hunting period. However, the determinants of such behaviour have not yet been studied. Herein, we investigate whether these non-permanent explorations are dispersal attempts/preludes or, in contrast, whether they are triggered by other factors such as disturbances related to hunting. Results Contrary to natal dispersal, we did not find strong male-bias in the propensity to engage in explorations. Exploration forays occurred less in juveniles than in adults and later in the season than natal dispersal. This was the case both for philopatric movements and for movements occurring after dispersal and settlement. These movements were also more likely to occur during the hare hunting period and the mating season. Conclusions We suggest that explorations in hares are triggered by factors other than dispersal and that hares may respond to hunting disturbances. Overall, we emphasize the need to account for human-related predation risk as a factor driving space-use in harvested species.

The bioluminescently marked Pseudomonas fluorescens strain 5RL, has been used previously to follow colonisation of soy bean roots (De Weger et al. [1991] Appl. Environ. Microbiol. 57:36-41). In the present paper the method has been further developed and optimized for wheat roots and it is used to get a quick overview of the colonisation patterns of many different root systems at the same time. Colonisation was followed on wheat plants grown in our gnotobiotic sand system (Simons et al., 1996. Mol Plant Microbe Interact 9: 600–607) and the following results were obtained. (i) A spatio-temporal analysis of the colonisation of wheat roots showed that 4 days after planting the highest bacterial activity was observed at the upper part of the root. After 6 days the high bacterial activity at the upper part was further increased, whereas spot-like activities were observed on the lower root parts, possibly due to micro-colonies. (ii) Bacterial mutations causing lack of motility or auxotrophy for amino acids resulted in impaired colonisation of the lower root parts, indicating that motility and prototrophy for the involved amino acid(s) are important factors for wheat root colonisation by strain 5RL. (iii) Coinoculation of strain 5RL with other wild type Pseudomonas strains on the root influenced the colonisation pattern observed for strain 5RL. Colonisation was not visually affected when the competing strain was a poor root coloniser, but was severely reduced when the competing strain was a good root coloniser. The results show that the spatio-temporal colonisation of wheat root by P. fluorescens strain 5RL and derivatives is similar to that of strain WCS365 on tomato. The advantage of the use of lux-marked strains is that the results are obtained much quicker than when conventional methods are used and that the result is supplied as an image of the colonisation pattern of many different roots.

Pollen monitoring is an important and widely used tool in allergy research and creation of awareness in pollen-allergic patients. Current pollen monitoring methods are microscope-based, labour intensive and cannot identify pollen to the genus level in some relevant allergenic plant groups. Therefore, a more efficient, cost-effective and sensitive method is needed. Here, we present a method for identification and quantification of airborne pollen using DNA sequencing. Pollen is collected from ambient air using standard techniques. DNA is extracted from the collected pollen, and a fragment of the chloroplast gene trnL is amplified using PCR. The PCR product is subsequently sequenced on a next-generation sequencing platform (Ion Torrent). Amplicon molecules are sequenced individually, allowing identification of different sequences from a mixed sample. We show that this method provides an accurate qualitative and quantitative view of the species composition of samples of airborne pollen grains. We also show that it correctly identifies the individual grass genera present in a mixed sample of grass pollen, which cannot be achieved using microscopic pollen identification. We conclude that our method is more efficient and sensitive than current pollen monitoring techniques and therefore has the potential to increase the throughput of pollen monitoring.

Repoblaciones de conejo europeo: una revisión crítica según las directrices de la IUCN (1998) para las reintroducciones Las repoblaciones de conejo europeo son una de las medidas más empleadas en los cotos de caza y en los proyectos de conservación en España, Francia y Portugal, donde el conejo es una especie clave. El objetivo de este trabajo consiste en revisar el conocimiento actual sobre los factores que afectan al establecimiento de las poblaciones de conejo reintroducidas según las directrices de la IUCN (1998), a fin de determinar las lagunas de conocimiento en este ámbito y destacar las técnicas que mejoran los buenos resultados reales de las reintroducciones. Ocho de los 17 puntos seleccionados de estas directrices se identificaron como desconocidos o parcialmente estudiados, incluidos importantes aspectos como el manejo y la liberación de conejos salvajes criados en cautividad, la elaboración de planes de transporte y seguimiento, la aplicación de programas de vacunación y los estudios a largo plazo posteriores a la liberación. Por lo tanto, los investigadores deben concentrar sus esfuerzos en suprimir esta falta de conocimiento y los gestores deben analizar todos los factores que aquí revisamos, con el objetivo de establecer unas directrices precisas para las futuras repoblaciones de conejo., European rabbit restocking is one of the most frequent actions in hunting estates and conservation projects in Spain, France and Portugal where rabbit is a keystone species. The aim of this work was to review current knowledge regarding rabbit restocking in accordance with the IUCN (1998) guidelines for re–introduction in order to identify gaps in knowledge and highlight the techniques that improve the overall success rate. Eight of 17 items selected from these guidelines were identified as partly studied or unknown, including important items such as the management and release of captive–reared wild rabbits, the development of transport and monitoring programs, the application of vaccine programs, and post–release long–term studies. Researchers should therefore concentrate their efforts on bridging these knowledge gaps, and wildlife managers should consider all the factors reviewed herein so as to establish accurate management guidelines for subsequent rabbit restocking programs.

Efficient colonization of the plant root is thought to be crucial for the plant-beneficial effect of particular Pseudomonas strains. Since root colonization is often the limiting step for successful plant growth stimulation, this process needs improvement. It is therefore important to acquire more information as regards (i) the conditions in the rhizosphere, and (ii) the bacterial traits that are involved in colonization. This review discusses some recent studies that focus on these two issues.

The impact of pollen on human health is primarily evident in allergic diseases. Sensitized patients can respond to pollen by symptoms of nose, eyes and bronchi. Pollen threshold levels for sensitization are unknown; instead most studies focus on the prevalence of sensitization for different pollen species. The pollen thresholds for symptom development vary among the different studies. Factors that influence the threshold level of a pollen species for symptom development are discussed. (i) Differences in response are observed among individual patients, but also among (ii) ethnic populations, (iii) changes in response to pollen concentrations during the pollen season occur, (iv) the amount of allergens carried by the pollen grains can differ in per region, from day to day and from year to year, and finally (v) threshold levels are affected by environmental factors, like weather conditions (temperature, pressure and storms), and air pollutants.The diversity of factors that influence the health impact of pollen has hampered the definition of a straight forward relationship between pollen and the severity of symptoms. However, within the public, the policymakers and the pharmaceutical industry there is a need for a definition of threshold pollen levels. A first approach to meet this need could be to define preliminary threshold values for different regions, followed by a validation of these preliminary threshold levels with patient symptom scores that can be collected by using new information and communication technology (ICT).