Es merged in physiological answer (NaCl 0.9) to prevent the drying dles (Figure 3). 9-Amino-6-chloro-2-methoxyacridine Chemical Because the as the abdominal cavity was opened, the abdomen was in (Figure 3). As quickly assoonabdominal cavity was opened, the abdomen was submergedsub- out organs. physiological reLacto-N-biose I Data Sheet solution (NaCl 0.9) to prevent theto avert the the internal organs.internal merged in physiological option (NaCl 0.9) drying out of drying out of theorgans.Figure 3. Queen abdominal cavity immediately after dissection (P6 in the figure was a note for tracking the sample identity).two.five. Spermatheca Extraction and AnalysisFigureAs Queen abdominal cavity right after dissection (P6 in the figurewas extracted and put a note 3. 3. Queen abdomen was opened, the spermatheca was the figure was the Figure soon because the abdominal cavity following dissection (P6 within a note for tracking on a sample slide. In 2017, the spermatheca was photographed on a slide beneath the microscope, sample identity). glass identity).for2.five. Spermatheca Extraction and AnalysisAs quickly as the abdomen was opened, the spermatheca was extracteAnimals 2021, 11, Animals 2021, 11, x6 ofand its diameter (ds) was measured with ImageJ software program. In 2018, the diameters As quickly because the abdomen was opened, the spermatheca was extracted and place on a measured by hand with a digital caliper. Because the spermatheca of a queen doesn’t glass slide. In 2017, the spermatheca was photographed on a slide beneath the microscope, a perfect spherical shape, the diameter was calculated because the diameters have been and its diameter (ds) was measured with ImageJ application. In 2018,the typical of three mea ments. by hand having a digital caliper. Given that (vs) was calculated as the volume of measuredThe volume on the spermatheca the spermatheca of a queen does not have aa spher perfectTo assess the concentration of calculated as the average of 3 measurements. mL of spherical shape, the diameter was semen, the spermatheca was popped in 1 The volume solution (NaCl 0.9). The concentration of sperm (sp) was assessed us iological of the spermatheca (vs) was calculated as the volume of a sphere. To assess the concentration of semen, (Burker Camera) below a in 1 mL of physstandard haemocytometer chamber the spermatheca was popped light microscope [48].iological remedy (NaCl 0.9). The concentration of sperm (sp) was assessed making use of a typical haemocytometer chamber (Burker Camera) under a light microscope [48].two.five. Spermatheca Extraction and Analysis2.6. Ovarioles Count2.6. Ovarioles the ovarioles with the correct ovary had been counted [37]. The amount of ovariol Only Count Only the ovarioles of your correct ovary had been counted [37]. et number of ovarioles was estimated with a strategy derived from PorporatoTheal. [46]. The ideal ovary w (o) was estimated using a approach derived from Porporato et al. [46]. The right ovary moved from the abdominal cavity and kept in staining solution for ca. 10 min (meth was removed in the abdominal cavity and kept in staining answer for ca. 10 min Blue Trihydrate, 0.5 , 0.five , Sigma-Aldrich, St. Luis, MO, USA). Soon after dying, the (methylene Blue Trihydrate,Sigma-Aldrich, St. Luis, MO, USA). After dying,the ovary washed washed with physiological remedy, placed on a slide, and analyzed beneath a ovary was with physiological option, placed on a slide, and analyzed beneath a stereom scope. The ovarioles have been separated using the the support of a dissecting needle and stereomicroscope. The ovarioles were separated withhelp of a dissecting needle and counte counted one particular by one particular,cut.