Fig 1.
Genital plugs of Urophonius species.
(A) Genital plug of Urophonius brachycentrus. (B) Genital plug of Urophonius achalensis, note that the plug is shown severed under the distal zone due to handling during extraction. Abbreviations: Dz, distal zone; Mz, middle zone; Pz, proximal zone. Scale bars: 1 mm.
Fig 2.
Hemocytes cells and distal substances found on the implants of females of Urophonius.
(A)(B) Surface of the genital implant of Urophonius brachycentrus under an optical microscope, note hemocyte cells. (C)(D) External female genitalia of Urophonius achalensis, note the genital implant and some surrounding substance in the area of the genital aperture. Abbreviations: bp, basal piece; ds, distal substance produced by the female; gi, genital implant; GR, granulocyte; ME, melanotic encapsulation; og, genital operculum; p, pectine; PL, plasmatocyte. Scale bars: A = 0.2 mm; B = 40 μm; C, D = 1 mm.
Fig 3.
Graphs of local vs systemic immunological parameters in females of two scorpion species.
Boxplots showing the distribution of data set and differences between the species and the type of response elicited (systemic: implants inserted in the body cavity; local: implants inserted in the genitalia) in females of Urophonius brachycentrus and Zabius fuscus. (A) Area of melanotic encapsulation (ME) (mm2). (B) Coloration of ME encapsulation (average grayscale value). Numbers above the boxplots indicate significant differences (p <0.05) between types of immune response elicited.
Fig 4.
Graphs of local immunological parameters in females of three scorpion species.
Top boxplots showing distribution of data set and differences between species, below heat maps charts in which average values are represented by colors (scale of reference of each variable to the side of the graph) according to the zones of the genital implant. (A) Area of melanotic encapsulation (ME) (mm2). (B) Area of non-melanotic encapsulation (NME) (mm2). (C) Coloration of NME encapsulation (average grayscale value). (D) Coloration of ME encapsulation (average grayscale value). Numbers indicate significant differences (p<0.05) between species above the boxplots. Letters on heat maps charts indicate significant differences (p <0.05) between zones of the genital implant within each species (i.e., intraspecific comparison: reading vertically, not horizontally).
Fig 5.
Hemocytes present in the hemolymph of females of three scorpions’ species.
(A) Plasmatocytes (PLs) and granulocytes (GRs) cluster of Urophonius brachycentrus. (B) GRs of Urophonius brachycentrus, note isodiametric morphology, granules in the cytoplasm and short and acute cytoplasmic extensions. (C) GRs of Zabius fuscus, note granules in the cytoplasm and short cytoplasmic extensions. (D) PL cluster of Urophonius achalensis. (E) PLs of Zabius fuscus expanding their cytoplasmic extensions with small inclusions in their cytoplasm. (F) Signet-ring PL of Urophonius brachycentrus, note vacuole occupied a large portion of the cytoplasm of the cell. Abbreviations: ce, cytoplasmatic extensions; GR, granulocyte; gs, granules in the cytoplasm; PL, Plasmatocyte; sg, signet-ring plasmatocyte; va, vacuole. Scale bars: A, D = 50 μm; B, C = 20μm; E, F = 10 μm.
Fig 6.
Total Hemocyte Load of females of the three scorpion species.
Differences in total hemocyte load (THL) (number of hemocytes per milliliter of hemolymph) according to the stage of quantification (pre-implantation and post-implantation in genitalia) of three scorpion species. Letters indicate significant differences (p<0.05) between species. Values are shown in scientific E-notation where 'E' represents the exponential to 10.
Fig 7.
Correlation matrix between the individual’s local immunological parameters and body weight.
In the diagonal portion of the matrix are found the parameters that were correlated with their corresponding frequency histogram: total hemocyte Load (THL) (number of hemocytes per milliliter of hemolymph h/ml); area of melanotic encapsulation (ME) (mm2); area of non-melanotic encapsulation (NME) (mm2); coloration of melanotic encapsulation (ME average grayscale value); coloration of non-melanotic encapsulation (NME average grayscale value); body weight (g). In the upper portion of the matrix are the Spearman correlation coefficients for each pair of variables and below the correlation p-value. In the lower portion of the matrix are graphs of dispersion of the variables, the colors of points represent the different species of scorpions (Black dots: Urophonius achalensis; Grey dots: Urophonius brachycentrus; White dots: Zabius fuscus). Abbreviations: ME, melanotic encapsulation of genital implant; NME, non-melanotic encapsulation of genital implant; THL, total hemocyte load. Some values are shown in scientific E-notation where 'E' represents the exponential to 10.