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Walls M.,Fertility Specialists of WA | Zuvela E.,Fertility Specialists of WA | Ayres C.,Demeter Laboratories | Sherrin D.,Queensland Fertility Group | And 8 more authors.
Asian Pacific Journal of Reproduction | Year: 2012

Objective: To undertake a multi-centre study to maximize the number of Makler chambers used. Methods: A total of 15 laboratories participated with 31 Makler chambers. A suspension of latex beads was prepared to a concentration of 20 millions per milliliter, and 0.5 mL aliquots distributed to each participating laboratory. They measured the concentration on their Makler chamber(s) used for routine semen analysis by adding 3, 4, 5, 7 and 10 μL volumes of bead suspension to the chamber. Results: There was no difference in within-chamber analysis of the bead concentration according to the volume of bead suspension applied within the range of 3-10 μL (F4,14=2.634, P=0.056). However, the between-chamber effects were significantly different (F30,124=4.937, P=0.000), and 24/31 (77.5%) chambers tested had an average bias>10% compared to the target bead concentration. Conclusions: A volume of 3-10 μL added to Makler counting chambers does not influence the concentration measured of latex beads, but the between-chamber variability and positive bias seen would suggest that other sources of error are present which are yet to be identified. © 2012 Hainan Medical College. Source

Pelzer E.S.,Queensland University of Technology | Pelzer E.S.,The Wesley Research Institute | Allan J.A.,Wesley Monash IVF | Waterhouse M.A.,The Wesley Research Institute | And 3 more authors.
PLoS ONE | Year: 2013

Our previous study reported microorganisms in human follicular fluid. The objective of this study was to test human follicular fluid for the presence of microorganisms and to correlate these findings with the in vitro fertilization (IVF) outcomes. In this study, 263 paired follicular fluids and vaginal swabs were collected from women undergoing IVF cycles, with various causes for infertility, and were cultured to detect microorganisms. The cause of infertility and the IVF outcomes for each woman were correlated with the microorganisms detected within follicular fluid collected at the time of trans-vaginal oocyte retrieval. Microorganisms isolated from follicular fluids were classified as: (1) 'colonizers' if microorganisms were detected within the follicular fluid, but not within the vaginal swab (at the time of oocyte retrieval); or (2) 'contaminants' if microorganisms detected in the vagina at the time of oocyte retrieval were also detected within the follicular fluid. The presence of Lactobacillus spp. in ovarian follicular fluids was associated with embryo maturation and transfer. This study revealed microorganisms in follicular fluid itself and that the presence of particular microorganisms has an adverse affect on IVF outcomes as seen by an overall decrease in embryo transfer rates and pregnancy rates in both fertile and infertile women, and live birth rates in women with idiopathic infertility. Follicular fluid microorganisms are a potential cause of adverse pregnancy outcomes in IVF in both infertile women and in fertile women with infertile male partners. © 2013 Pelzer et al. Source

Pelzer E.S.,Queensland University of Technology | Pelzer E.S.,The Wesley Research Institute | Harris J.E.,Queensland University of Technology | Allan J.A.,Wesley Monash IVF | And 4 more authors.
Journal of Reproductive Immunology | Year: 2013

Recently we reported the presence of bacteria within follicular fluid. Previous studies have reported that DNA fragmentation in human spermatozoa after in vivo or in vitro incubation with bacteria results in early embryo demise and a reduced rate of ongoing pregnancy, but the effect of bacteria on oocytes is unknown. This study examined the DNA within mouse oocytes after 12 hours' incubation within human follicular fluids (. n=. 5), which were collected from women undergoing in vitro fertilization (IVF) treatment. Each follicular fluid sample was cultured to detect the presence of bacteria. Terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL) was used to label DNA fragmentation in ovulated, non-fertilized mouse oocytes following in vitro incubation in human follicular fluid. The bacteria Streptococcus anginosus and Peptoniphilus spp., Lactobacillus gasseri (low-dose), L. gasseri (high-dose), Enterococcus faecalis, or Propionibacterium acnes were detected within the follicular fluids. The most severe DNA fragmentation was observed in oocytes incubated in the follicular fluids containing P. acnes or L. gasseri (high-dose). No DNA fragmentation was observed in the mouse oocytes incubated in the follicular fluid containing low-dose L. gasseri or E. faecalis. Low human oocyte fertilization rates (<29%) were associated with extensive fragmentation in mouse oocytes (80-100%). Bacteria colonizing human follicular fluid in vivo may cause DNA fragmentation in mouse oocytes following 12. h of in vitro incubation. Follicular fluid bacteria may result in poor quality oocytes and/or embryos, leading to poor IVF outcomes. © 2013 Elsevier Ireland Ltd. Source

Pelzer E.S.,Queensland University of Technology | Allan J.A.,Wesley Monash IVF | Cunningham K.,Queensland University of Technology | Mengersen K.,Queensland University of Technology | And 4 more authors.
Human Reproduction | Year: 2011

Background: Previous studies have measured cytokines expressed within follicular fluid and compared the profiles with the aetiology of infertility and/or successful or unsuccessful assisted reproduction technology (ART) outcomes. Methods: In this study, 71 paired follicular fluid and vaginal secretions collected from ART patients were cultured to detect microorganisms and tested for the presence of cytokines. Patient specimens were selected for assay based on two criteria: whether the follicular fluid specimen was colonized (with microorganisms prior to oocyte retrieval) or contaminated by vaginal flora and; the aetiology of infertility. Patients included fertile women (with infertile male partners; n 18), women with endometriosis (n 16) or polycystic ovary syndrome (PCOS, n 14), or couples with a history of genital tract infection (n 9) or idiopathic infertility (n 14). Results: Microorganisms and cytokines were detected within all tested specimens. Colonizing microorganisms in follicular fluid were associated with: decreased fertilization rates for fertile women (P 0.005), women with endometriosis (P 0.0002) or PCOS (P 0.002) compared with women whose follicular fluid was contaminated at the time of oocyte retrieval and with decreased pregnancy rates for couples with idiopathic infertility (P 0.001). A single cytokine was discriminatory for women with an idiopathic aetiology of infertility (follicular fluid interleukin (IL)-18). Unique cytokine profiles were also associated with successful fertilization (IL-1α, IL-1β, IL-18 and vascular endothelial growth factor). Conclusions: Follicular fluid is not sterile. Microorganisms colonizing follicular fluid and the ensuing cytokine response could be a further as yet unrecognized cause and/or predictor of adverse ART outcomes and infertility. © 2011 The Author. Source

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