PMC10203518_01
Timeline of the clinical evolution, diagnostic studies, pathogen identification, therapy and outcome of the patient case study.
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F1
A 36-year-old male presented to a Urology and Andrology outpatient clinic of a healthcare center from Cordoba, Argentina, in July 2018 referring signs and symptoms of epididymo-orchitis (Figure 1). The patient's medical record revealed a history of varicocele surgery 1 year earlier (July 2017) (Figure 1). Table 1, a positive result was obtained for C. trachomatis infection while all the other pathogens were negative. The urogenital infection by C. trachomatis was later confirmed on August 17th 2018 in an independent sample (Table 1, Supplementary Figure 1, Figure 1). All these findings allowed the diagnosis of chronic epididymitis due to C. trachomatis infection and a treatment with doxycycline 100 mg/12 h for 45 days was prescribed (Figure 1). Negative results were obtained for C. trachomatis as well as for all the other pathogens investigated indicating resolution of the infection (Table 1, Supplementary Figure 1, Figure 1).
Testicular color Doppler ultrasounds showing hyperechogenicity and enlarged right epididymis with markedly increased vascularity that extended to the fatty spermatic cord compatible with epididymitis (A) and funiculitis (B) at patient admission (indicated with arrows), and almost complete resolution of tissue changes after antibiotic therapy and infection clearing (C). Assessment of sperm viability by flow cytometry using propidium iodide staining as previously described (D). Representative histograms depicting frequencies of necrotic spermatozoa (propidium iodide-positive) within the spermatozoa population gated on FSC vs. SSC plots. Data were collected on FACS-CANTO II flow cytometer (BD Biosciences, San Diego, USA) and analyzed using FlowJo software (version 7.6.2). Proper compensation using Fluorescence Minus One (FMO) controls were used. Quantification of reactive oxygen species (ROS) level in seminal plasma by chemiluminescence using luminol (Sigma-Aldrich, St. Louis, USA) was carried out according to the WHO laboratory manual for the examination and processing of human semen (E). Measurements were made with use of a Berthold luminometer (model LKB 953; Wallac Inc., Gaithersburg, USA). Results are expressed as percentages (%) or counted photons per minute (cpm). Experiments were performed in triplicates. Data are shown as mean +- SD. Mann-Whitney test; *p<0.05.
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F2
These findings were consistent with focal epididymitis and funiculitis (Figures 2A, B). A post-treatment Doppler ultrasound showed scarce to absent hypervascularization and slightly increased size and echogenicity of the right epididymitis, findings that were compatible with sequelae of focal chronic epididymitis (Figure 2C). In addition, no signs of inflammation in the right spermatic cord indicated resolution of the funiculitis (Figure 2C). Moreover, the resolution of the infection associated with significant reductions in ROS and sperm necrosis levels (Figures 2D, E). MLST allelic profile showed an exact match with C. trachomatis sequence type 141 (ST141). Moreover, the ompA gene sequence obtained was 100% identical to that of C. trachomatis genovar L2, confirming that the strain detected was C. trachomatis LGV L2 (Supplementary Figure 2 and Supplementary Table 2).