Abstract

Simple SummaryIn animal production, the prediction of male fertility is vital for the success of specific techniques such as artificial insemination. Thus, testicular ultrasound, a non-invasive diagnostic procedure, could be a useful tool. Moreover, recent ultrasound-video analysis and software developments allow the visualization of tissue at the microscopic level. The objective of this work was to establish a possible correlation between testicular ultrasonography and semen quality in rams. For this purpose, the testicles of nine rams were evaluated and the semen was analyzed for one year. The results revealed that the number of white and grey pixels correlated with sperm parameters indicating poor seminal quality. On the other hand, the increase in the seminiferous-tubule density or the lumen area of these tubules was related to a rise in seminal quality. Therefore, ultrasound-video analysis could be a good tool for evaluating the fertility of rams, either for artificial insemination or on the farm.Testicular ultrasound is a non-invasive technique that could be very useful for predicting ram seminal quality. Recent software developments allow macroscopic and microscopic evaluation of testicular parenchyma. Thus, the aim of this study was to evaluate the testicular echotexture using ultrasound-video analysis and investigate its possible correlation with semen quality. Nine rams were evaluated for one year using a portable ultrasound scanner and the echotexture was analyzed with ECOTEXT® software. The number of black (Ec1), white (Ec2), and grey pixels (Ec3), tubular density (TD), lumen area (LA), and lumen diameter (LD) were analyzed. Semen was collected by an artificial vagina the same day and the sperm concentration, morphology, motility, viability, phosphatidylserine (PS) translocation, reactive-oxygen-species (ROS) levels, DNA damage and capacitation state were evaluated. Ec2 and Ec3 correlated positively with “bad quality” sperm parameters (the percentage of spermatozoa with high ROS levels, with PS translocation and proximal cytoplasmic droplets), and negatively with motility. In contrast, TD and LA showed a positive correlation with “good quality” parameters (motility or normal morphology) and a negative correlation with spermatozoa with high ROS levels, with DNA fragmentation, and proximal or distal cytoplasmic droplets. Thus, echotexture analysis by ultrasound-video analysis could be a valuable tool for assessing ram fertility.

Highlights

  • Seasonality, which is regulated by photoperiod and melatonin secretion [1], could be a limiting factor in the sheep reproduction [2]

  • Of the Doppler parameters, only the frequency significantly increased (p < 0.05) during the non-breeding season (Table 1), a non-significant decrease in Pulsatility Index (PI) was observed in the coldest months of the year (Supplemental Figure S1B)

  • Previous studies had shown an increase in testicular numerical-pixel values (NPVs) and pixel heterogeneity during the non-breeding seasons (NB) season [15,16] that could have been related to changes in the seminiferous tubules [13,14]

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Summary

Introduction

Seasonality, which is regulated by photoperiod and melatonin secretion [1], could be a limiting factor in the sheep reproduction [2]. A dedicated software package has been developed for studying testicular tissue through macroscopic and microscopic ultrasound-video analysis (Ecotext® software) This software is able to analyze testicle ultrasonograms based on their echotexture properties, measuring pixel intensity as Ec1 (black pixels, the number of pixels with NPV = 0), Ec2 (white pixels, the number of pixels with NPV = 255), and Ec3 (grey level of pixels, the mean value of the pixels with NPV > 0 and

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