Implementing an open-access CASA software for the assessment of stallion sperm motility: Relationship with other sperm quality parameters.

In the field of assisted reproductive techniques (ART), computer-assisted sperm analysis (CASA) systems have proved their utility and potential for assessing sperm quality, improving the prediction of the fertility potential of a seminal dose. Although most laboratories and scientific centers use commercial systems, in the recent years certain free and open-source alternatives have emerged that can reduce the costs that research groups have to face. However, these open-source alternatives cannot analyze sperm kinetic responses to different stimuli, such as chemotaxis, thermotaxis or rheotaxis. In addition, the programs released to date have not usually been designed to encourage the scalability and the continuity of software development. We have developed an open-source CASA software, called OpenCASA, which allows users to study three classical sperm quality parameters: motility, morphometry and membrane integrity (viability) and offers the possibility of analyzing the guided movement response of spermatozoa to different stimuli (useful for chemotaxis, thermotaxis or rheotaxis studies) or different motile cells such as bacteria, using a single software. This software has been released in a Version Control System at Github. This platform will allow researchers not only to download the software but also to be involved in and contribute to further developments. Additionally, a Google group has been created to allow the research community to interact and discuss OpenCASA. For validation of the OpenCASA software, we analysed different simulated sperm populations (for chemotaxis module) and evaluated 36 ejaculates obtained from 12 fertile rams using other sperm analysis systems (for motility, membrane integrity and morphology modules). The results were compared with those obtained by Open-CASA using the Pearson’s correlation and Bland-Altman tests, obtaining a high level of correlation in all parameters and a good agreement between the different used methods and the OpenCASA. With this work, we propose an open-source project oriented to the development of a new software application for sperm quality analysis. This proposed software will use a minimally centralized infrastructure to allow the continued development of its modules by the research community.

/pdf/opencasa-a-new-open-source-and-scalable-tool-for-sperm-30a6t7qa7l.pdf

OpenCASA: A new open-source and scalable tool for sperm quality analysis

Swim-up is a sperm purification method that is being used daily in andrology labs around the world as a simple step for in vitro sperm selection. This method accumulates the most motile sperm in the upper fraction and leaves sperm with low or no motility in the lower fraction. However, the underlying reasons are not fully understood. In this article, we compare metabolic rate, motility and sperm tail length of bovine sperm cells of the upper and lower fraction. The metabolic assay platform reveals oxygen consumption rates and extracellular acidification rates simultaneously and thereby delivers the metabolic rates in real time. Our study confirms that the upper fraction of bull sperm has not only improved motility compared to the cells in the lower fraction but also shows higher metabolic rates and longer flagella. This pattern was consistent across media of two different levels of viscosity. We conclude that the motility-based separation of the swim-up technique is also reflected in underlying metabolic differences. Metabolic assays could serve as additional or alternative, label-free method to evaluate sperm quality.

/pdf/the-motility-based-swim-up-technique-separates-bull-sperm-3du4h0r9vy.pdf

The motility-based swim-up technique separates bull sperm based on differences in metabolic rates and tail length.

Development of computer-assisted sperm analysis plugin for analyzing sperm motion in microfluidic environments using Image-J.

For reproduction, most fish species adopt external fertilization: their spermatozoa are delivered in the external milieu (marineor freshwater) that represents both a drastic environment and a source of signals that control the motility function. This chapter is an updated overview of the signaling pathways going from external signals such as osmolarity and ionic concentration and their membrane reception to their transduction through the membrane and their final reception at the flagellar axoneme level. Additional factors such as energy management will be addressed as they constitute a limiting factor of the motility period of fish spermatozoa. Modern technologies used nowadays for quantitative description of fish sperm flagella in movement will be briefly described as they are more and more needed for prediction of the quality of sperm used for artificial propagation of many fish species used in aquaculture. The chapter will present some applications of these technologies and the information to which they allow access in some aquaculture species.

/pdf/fish-sperm-physiology-structure-factors-regulating-motility-1ee0b1ec1c.pdf

Fish Sperm Physiology: Structure, Factors Regulating Motility, and Motility Evaluation

Summary

Computer assisted sperm analysis (CASA) has greatly enhanced the quality and quantity of data that can be collected on spermatozoa movements. Open source imagej software allows any researcher to have access to such analyses, but CASA systems in general are not designed to ably process large numbers of video sequences. Here we introduce a modified imagej CASA_automated plugin that fundamentally enhances efficiency and will allow higher quality research to be conducted with no extra post-microscope effort.

Modifications to the imagej computer assisted sperm analysis plugin greatly improve efficiency and fundamentally alter the scope of attainable data

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Modifications to the imagej computer assisted sperm analysis plugin greatly improve efficiency and fundamentally alter the scope of attainable data