ASSISTED REPRODUCTION TECHNOLOGIES
Increased count, motility, and total motile sperm cells collected across three consecutive ejaculations within 24 h of oocyte retrieval: implications for management of men presenting with low numbers of motile sperm for assisted reproduction
Al-Hasen Said1 & Michael L. Reed1
Received: 25 February 2015 /Accepted: 2 June 2015 # Springer Science+Business Media New York 2015
Purpose The purpose of this study was to quantitate changes in seminal volume, sperm count, motility, qualitative forward progression, and total motile sperm cells per ejaculate, across three consecutive ejaculates collected from individuals within 24 h preceding an IVF cycle.
Methods Men presenting with oligoasthenozoospermia or asthenozoospemia attempted three ejaculates within 24 h preceding IVF. Ejaculate 1 was produced the afternoon prior to oocyte retrieval, and ejaculates 2 and 3 were produced the morning of oocyte retrieval with 2–3 h between collections.
Ejaculates 1 and 2 were extended 1:1 v/v with room temperature rTYBS. Test tubes were placed into a beaker of room temperature water, then placed at 4 °C for gradual cooling.
Ejaculate 3 was not extended, but pooled with ejaculates 1 and 2 and processed for intracytoplasmic sperm injection (ICSI).
Results Out of 109 oocyte retrievals, 28 men were asked to attempt multiple consecutive ejaculations. Among this population, 25/28 (89.3 %) were successful, and 3/28men (10.7 %) could only produce two ejaculates. Mean volumes for ejaculates 1, 2, and 3 were significantly different from each other (p<0.01); the volume decreased for each ejaculate. Mean sperm counts, motility, qualitative forward progression, and total motile cells per ejaculate for the ejaculates1, 2, and 3 demonstrated the following: ejaculates 2 and 3 were not significantly different, but counts, motility, and total motile sperm were improved over ejaculate 1 (p<0.01).
Conclusions Pooling three consecutive ejaculates within 24 h increased the numbers of available motile sperm in this population by 8-fold compared to the first ejaculate alone, facilitating avoidance of sperm cryopreservation and additional centrifugation steps that could affect sperm viability and/or function.
Keywords IVF . ICSI . Sexual abstinence .
Oligoasthenozoospermia . Fertility preservation
In our laboratory, men presenting with oligozoospermia, asthenozoospermia, and oligoasthenozoospermia  are asked to attempt three consecutive ejaculates within 24 h of the scheduled oocyte retrieval: one ejaculate to be attempted mid-afternoon prior to the retrieval and two ejaculates attempted the morning of the retrieval, within a period of 2 to 3 h. The three consecutive ejaculates are pooled prior to processing for IVF. The goals of this novel strategy were to alleviate stress-induced anejaculation on the day of oocyte retrieval [2, 3] and to maximize quality and numbers of motile sperm available by eliminating potential iatrogenic cryopreservation-induced damage, further loss of alreadydiminished numbers of motile sperm following cryopreservation, and eliminating unpredictability of between- and withinpatient post-thaw sperm viability [4–7]. And, while we stipulate that in most, if not all cases, there would likely have been sufficient sperm in a single ejaculate from these men for ICSI,
Capsule Pooling consecutive ejaculates produced within 24 h significantly increased the numbers of available motile sperm for IVF in asthenozoospermic and oligoasthenozoospermic men by 8-fold over a single ejaculate alone. * Michael L. Reed email@example.com
Alh.firstname.lastname@example.org 1 Center for Reproductive Medicine of NewMexico, 201 Cedar Street
SE, Suite S1-20, Albuquerque, NM 87106, USA
J Assist Reprod Genet
DOI 10.1007/s10815-015-0509-z we considered it very important to avoid potentially damaging effects of centrifuge-mediated sperm pelleting, e.g., generation of reactive oxygen species, by being able to process sperm by gradient separation [8–11].
Over time, we noted, but failed to quantify, a pattern of increased sperm numbers and/or motility in the second and/ or third ejaculations of most men. This phenomenon is not widely documented [12–14], but it has been discussed anecdotally between laboratory professionals where it has been a routine practice to ask for a second specimen, spontaneously, in the event of a poor first specimen.
In context to the semen analysis parameters for the patient population described in this study, oligozoospermic men may benefit from multiple consecutive, or more frequent ejaculations compared to normozoospermic men [12, 15, 16]. Also,
Koscinski et al.  trialed a multiple specimen collection strategy in an effort to avoid sending cryptozoospermic patients for testicular surgery—two ejaculates were collected approximately 2 h apart on the day prior to IVF, the specimens were pooled, washed, and briefly incubated in microdrops, after which motile sperm were captured using an injection pipette and moved to a separate microdrop. The motile sperm that survived overnight incubation were then used in some of the men for IVF with ICSI.
The goal of this observational, retrospective study was to quantitate and document changes in basic semen analysis parameters (volume, sperm count, motility, and qualitative forward progression) and calculated total motile sperm per ejaculate, across three ejaculates within individual patients, collected within approximately 24 h of planned oocyte retrieval.
The time frame designated for this study was one calendar year, and the null hypothesis was that frequent ejaculations over a short time frame (three ejaculations within 24 h) would not alter semen analysis parameters from ejaculate to ejaculate.
Materials and methods
A deidentified data base was examined retrospectively as a basis for the study, and therefore, it does not contain clinical or other identifiers that might disclose the identity of the patients. The methodology for semen collection, extension, and processing was standard operating procedure for our laboratory and not experimental; therefore, patient consent and Institutional Review Board approval were not solicited.