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. 2021 Feb 10;57(2):159. doi: 10.3390/medicina57020159

Table 4.

Evaluation procedure and results of sperm quality parameters.

Study Variable Measurement Procedure Results
Samadian 2019 [16] Cell count Analyses by light microscopic Figure 2
Apoptosis index Evaluated by TUNEL (Terminal deoxynucleotidyl transferase enzyme-mediated dUTP nick end labeling) staining kit (In situ cell death detection kit, POD, Roche; Cat, Germany). (Con = Ex = DiaIn = DiaInEx) < DiaEx < Dia
Samadian 2019 [17] Quality Sperm: Count Incubation of tissues in heat transfer fluid (HTF) medium, 45 min at 37 °C in 5% CO2. Dilution of 1:50 and observation with light microscope stereo zoom microscope. Count based on standard Neobar slide method. Figure 2
Quality Sperm: Motility Incubation of tissues in HTF medium, 45 min at 37 °C in 5% CO2. Dilution of 1:50 50 and observation with light microscope stereo zoom microscope. Figure 3
Quality Sperm: Impaired chromatin condensation Aniline blue staining. The spermatozoa with unstained nuclei were considered as mature (representing protamine content), while the spermatozoa with stained nuclei (representing histone content) were marked as immature [22]. (Con = Ex) < (DiaIn = DiaInEx = DiaEx) < Dia
Quality Sperm: DNA Fragmentation Acridine orange Staining and evaluation by fluorescence microscope (Zeiss Company). Spermatozoa with green DNA content were marked as spermatozoa with double-stranded DNA, while the spermatozoa with yellowish and/or reddish fluorescent DNA content were considered as spermatozoa with DNA fragmentation. (Con = Ex) < (DiaIn = DiaInEx = DiaEx) < Dia
Parastesh 2019 [20] Sperm count (106) Dissected epididymis was transferred into a 5 cc (DMEM) medium. Sperm swim out into the medium. One mL of the solution was diluted with 9 mL formaldehyde fixative. Neubauer chamber and manually counted under a microscope. Figure 2
Sperm viability Eosin-nigrosine staining was used to evaluate sperm viability according to WHO protocol [23]. Ex (87.8 ± 2.9%) > Con (77.5 ± 4.6%) > DiaEx (41.7 ± 7.2%) > Dia (29.7 ± 16.2%)
Sperm morphology Papanicolaou stain according to WHO protocol [23]. Día (85.2 ± 7.5) < Con (95.4 ± 1.3);
Ex (90.4 ± 9.1) > (Dia (85.2 ± 7.5) = DiaEx (88 ± 8.8))
Sperm motility WHO protocol [23]. Ten µl of the sperm suspension was located on a microscope slide. Figure 3
Parastesh 2019 [21] Sperm count (106) Dissected epididymis was transferred into a 5 cc (DMEM) medium. Sperm swim out into the medium. 1 mL of the solution was diluted with 9 mL formaldehyde fixative. Neubauer chamber and manually counted under a microscope. Figure 2
Sperm viability Eosin-nigrosine staining was used to evaluate sperm viability according to WHO protocol [23]. (Dia (29.7 ± 16.2) = DiaExE (41.7 ± 7.2)) < Con (77.5 ± 4.6); DiaExR (60.6 ± 8) and DiaExC (60.6 ± 8) > Dia (29.7 ± 16.2)
Sperm morphology Papanicolaou stain according to WHO protocol [23]. Dia (85.2 ± 7.5) < Con (95.4 ± 1.3);
DiaExE (88 ± 8.8) = DiaExR (89.1 ± 6.9) = DiaExC (93.4 ± 2.7)
Sperm motility WHO protocol [23]. Ten µl of the sperm suspension was located on a microscope slide and covered. Figure 3
Rosety Rodríguez 2014 [19] Volume Semen was collected after 3 days of sexual abstinence (intercourse or masturbation) by manual masturbation into a sterile container on-site and examined within 30 min of ejaculation [23]. Dia (pre, 2.90 ± 0.86) = Dia (post, 2.92 ± 0.90);
DiaEx (pre, 2.92 ± 0.90 = DiaEx (post, 3.08 ± 0.86)
Concentration Hemacytometer (HauserScientific Inc., Horsham, PA, USA) Figure 2
Motility Computer-aided semen analysis (CASA system, Microptic S.L., Spain) Figure 3
Morphology Hemacytometer (Hauser Scientific Inc., Horsham, PA, USA) and Computer-aided semen analysis (CASA system, MicropticS.L., Spain) DiaEx (pre, 46.1 ± 4.6) < DiaEx (post, 51.2 ± 5.0)
DiaEx (post, 51.2 ± 5.0) > Dia (post, 45.9 ± 4.8)
Murray 1988 [18] Evaluation of sperm quality before exercise One single measure comparing diabetic and healthy young males:
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    Volume: Dia (2.25 ± 0.6); Con (2.98 ± 0.7)

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    Sperm density: Dia (41.5 ± 10.7); Con (57.4 ± 10.7)

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    Motility: Dia (46.3 ± 6.1); Con (53.5 ± 4.4)

Con: control group, Dia: diabetic group, DiaIn: insulin administration group, DiaEx: diabetic and exercise intervention group, DiaInEx: diabetic group with insulin administration and exercise intervention, DiaExE: diabetic and endurance exercise intervention group, DiaExR: diabetic and resistance exercise intervention group, DiaExC: diabetic and combined exercise intervention group, WHO: World Health Organization, HTF: heat transfer fluid.