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  • Specifications table br Value of the data br Experimental

    2018-10-23

    Specifications table
    Value of the data
    Experimental design, materials and methods The digital camera was attached to light transmission microscope with 10× objective magnification and no eye pieces. The blood smear has been illuminated by 50W halogen lamp and the image focused by normal microscopic focusing system in order to capture high resolution image of the stained blood smear. The acquired images subjected to segmentation process as a part of the date gap junctions stage involves the partitioning of the image plane into meaningful parts [1,2]. In this work a system scheme (Diagram 1) is proposed to segment both the RBCs form the background and the counting area which indicated by the triple line square on the glass slide [3]. The full description of the segmentation scheme is given as follows: At the first step, the captured smear image split into its three color component bands (red, green and blue) as shown in Fig. 1. The three grayscale components are examined and we note that the green components show a clear image of RBCs [4]. At the second step, the histogram green component has been performed and the initial threshold value taken as gray level corresponding to the peak of the histogram as shown in Fig. 2. At the third step, the areas of the objects on the threshold image has been determined which shows that the most common areas are less than 10 pixels corresponding to the area of the RBCs. The objects that have areas larger than this value (10) are due to inaccurate threshold as shown in Fig. 3. Based on this, if there is any object which has an area greater than 10 the feedback increases the value of threshold by one until no longer object has an area greater than 10 as shown in Fig. 4. At the fourth step, extracting the counting area by calibrating the cropped image to be the same area of the triple line square at that system setup as shown in Fig. 5. Finally, the number of objects in the extracted area determined which is corresponding to the number of RBCs in that area. A complete set of images, gap junctions their RBCs counts and segmentations are included as supplementary material to this article.
    Data, experimental design, materials and methods
    Acknowledgements Authors wish to thank the Finland Ministry of Foreign Affairs (CHIESA project [Climate Change Impacts on Ecosystem Services and Food Security in Eastern Africa]), French Ministry of Foreign Affairs and IRD (Institut de Recherche pour le Développement) (KENCCA-JEAI project [KENyan Climate Change and Adaptation-Jeunes Equipes AIRD]) and icipe (Nairobi, Kenya) for their financial supports. Authors wish to thank to Eric Muchugu for interpolation of rainfall data for Machakos Hills transect. Thanks are also given to Fritz Schulthess for his review of the manuscript and to the anonymous reviewers for their helpful comments and corrections.
    Data The data presented here show the prevalence of eucalypt pollen (percentage of individuals with eucalypt pollen) and of pollen of other types and the average pollen loads per individual for eucalypt and other pollen types for all bird species captured in a 14-month period, together with the number of captured individuals. See [1] for further information and discussion.
    Experimental design, materials and methods The study was carried out in two constant effort bird ringing stations operated by the bird ringing group Anduriña (see acknowledgments) in NW Spain (Darbo: 42°15′48.01″N 8°47′46.97″W; Coiro: 42°16′29.60″N 8°46′16.61″W). The stations are located 2.5km apart in mixed landscapes with agricultural land, abandoned fields, native forest patches, forestry plantations (of Eucalyptus globulus and Pinus pinaster) and scattered houses with their gardens. The two sites differ in the surface covered by E. globulus. In Coiro, E. globulus stands covered 19.5% of the area comprised within 300-m distance from mist nets (total area of 31.3ha), with a minimum distance between mist nets and E. globulus trees of c. 50m. In Darbo, E. globulus stands covered 7.9% (of an area of 17.7ha) and the minimum distance between mist nets and E. globulus trees was c. 200m. We set nine 12-m long nets in Darbo, making a total of 108m arranged in three 36-m long lines. In Coiro, we set six 12-m long nets, making a total of 76m arranged in four lines of 12m or 24m. Mist nets were operated one day per month in each site, from dawn to dusk, usually on two consecutive Saturdays: the closest to the 15th day of each month and the next one (weather permitting, avoiding rainy and windy days). The study was carried out between March 2014 and April 2015 (i.e. 14 months). For all birds captured, we collected pollen grains attached to the bill and surrounding feathers (forehead, chin and cheeks). For this, we used a lab spatula impregnated with a glycerine-based gel, which was then transferred to microscope slides [2]. When the amount of pollen was too much for one slide, we used two slides, or transferred a subsample of the whole amount to slides (1/2 or 1/4). Samples were processed by acetolysis [3] to aid pollen identification. The sampling effort was adjusted according to the density of pollen in each slide: for slides with low pollen density (≤2500 grains/cm2) pollen was quantified in 30 fields at ×100 magnification (19% of the area sampled; [4]), with further observation of the pollen grains detected in each field at ×400 magnification for identification. For slides with high pollen density (>2500 grains/cm2) we used 30 fields at ×400 magnification. Pollen was identified using our own pollen reference collections and the aid of expert palynologists (see acknowledgments).