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All experiments were repeated at least twice and each sample was evaluated in triplicate.
Each sample was evaluated for sperm ubiquitination by the direct immunofluorescence method, using anti-ubiquitin antibodies.
A minimum of 100 cells from each sample was evaluated for the presence or absence of membrane staining.
The amount of organic matter in each sample was evaluated using Loss On Ignition (LOI), a hydrogen peroxide (H2O2) treatment, and a sodium hypochlorite (NaClO) treatment following the H2O2 treatment.
The actual number of living haemocytes in each sample was evaluated using a Thoma counting chamber.
Each sample was evaluated in triplicate, and each experiment contained a negative control corresponding to the RLU values obtained with empty HIV-1 pseudoparticles.
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For the upper respiratory tract, 16 sections of each sample were evaluated.
The concentrations of free amino acids in each sample were evaluated using an automated L-8800A Acido Analyzerlyzer (Hitachi, Tokyo, Japan) according to a previously described method [33], [34], [35], [36].
At least four samples from each sample were evaluated.
Complementary DNAs of each sample were evaluated in triplicate.
The SNPs in each sample were evaluated for MAF and HWE.
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