Here is a question that has came up:

What is the difference between Chymotrypsin and Hyaluronidase?

Answer:

In the article: Stefan Stephanus du Plessis, Sheila Gokul, Ashok Agarwal. Frontiers in Bioscience, Elite, 5, 224-231, January 1, 2013, the following information on semen hyperviscosity (SHV) treatment can be found:
“Semen hyperviscosity can be treated successfully in vitro by traditional methods as well as by recently developed methods. Hyperviscous semen is commonly diluted or drawn into a hypodermic needle and forced through in order to overcome the elevated viscosity, although these methods are unlikely to be completely effective because SHV is not completely a mechanical phenomenon. More direct treatments of patients presenting with SHV include over hydration, prostatic massage, and the use of parenteral hyaluronidase (injection – my note). These methods showed limited success and have proven not to be too effective.

Less dated methods used in sperm preparation for ART procedures have been shown to improve semen and sperm parameters of infertile males. Honea et al. found that the use of limited proteolysis by using α-chymotrypsin in the treatment of SHV was effective for an in vitro setting such as IVF or IUI. Zavos et al. more recently reported that limited proteolysis by α-chymotrypsin was effective in improving the use and handling of hyperviscous semen samples, and that treatment with α-chymotrypsin assisted in the recovery of high quality sperm in greater numbers than in hyperviscous samples not treated with α-chymotrypsin.”  

Conclusions:

• Though hyaluronidase is present in semen and used in both IVF procedures and for treatment of patients (via injection), it is not recommended for semen hyperviscosity treatment in vitro because there is no evidence of effectiveness.
• α-chymotrypsin was found to be an effective method for SHV in vitro treatment based on the methods listed above and years of clinical practice. 
• The QwikCheck Liquefaction Kit (based on α-chymotrypsin) product insert is enclosed, demonstrating the effectiveness of the product in clinical studies.  

DOWNLOAD QWIKCHECK LIQUEFACTION KIT PRODUCT INSERT

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It is important to understand the relationship and difference between Precision and Accuracy.  Both are important, and both showcase different aspects of a Validation or Comparison study. The image below provides a very nice visual representation:

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Question:

In addition to the obvious issues with sample loading in the SQA testing capillary, would concentration readings be higher or lower in a sample which has not fully liquefied?

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Based on the required semen sample testing volume of 0.5 mL, it is recommended to fill the entire testing capillary with a semen sample without any dilution if the ejaculate volume is >= 0.5 mL. Sometimes though the ejaculate volume may be less than 0.5 mL. The dilution mode of the SQA-V and Vision can be used in this case as follows: If the semen sample volume is >= 0.3 mL, dilute the sample using the QwikCheck Dilution media two times (1:2 or 1+1). For example, 0.3 mL of semen sample is mixed with 0.3 mL of diluent resulting in a volume of 0.6 mL. This will provide an adequate sample volume to aspirate into the SQA testing capillary and the SQA software automatically compensates for the dilution and will provide a full report, no calculations are required by the operator.

RECOMMENDATIONS
Based on the sample volume, M.E.S. recommends the following approach to testing the sample:
• Semen Volume >= 0.5 mL - No dilution is required
• Semen Volume >= 0.3 mL and < 0.5 mL - Dilution 1:2 (1+1) using the QwikCheck Dilution kit
• Semen Volume < 0.3 mL - Manual assessment [microscope or Vision Manual Mode or Vision Semi-automated Mode (10 µL automated testing + 10 µL manual motility assessment)]

REFERENCES
• WHO laboratory manual for the examination and processing of human semen - 5th ed., WHO 2010.
• SQA-V GOLD User Guide Version 2.60 I-Button
• SQA-Vision User Guide Version 104.13.2

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