Animal Sciences

This study evaluated bovine amniotic and allantoic fluids as culture media for two-cell murine embryos to the hatched blastocyst stage. Amniotic and allantoic fluids were collected from four 70-d periods of pregnancy and pooled from at least five different animals. In Experiment 1 (n=470) the fluids were frozen twice. Treatments consisted of twice frozen amniotic or allantoic fluid from each pregnancy period, Whitten's medium and fetal calf serum. The later two media were controls. Twice-frozen amniotic fluid<70 d pregnancy period, fetal calf serum and Whitten's medium supported the development of embryos to the hatched blastocyst stage. Whitten's medium was superior to twice-frozen amniotic fluid<70 d pregnancy period or fetal calf serum (P<0.01). Biochemical analysis showed lower glucose in amniotic and allantoic fluids than in Whitten's medium. Experiment 2 (n=425) was performed to evaluate the effect of glucose supplementation to amniotic fluid. No benefit of glucose supplementation of the amniotic fluid was observed. In Experiment 3 (n=432), the fluids were transported nonfrozen on ice. Treatments consisted of nonfrozen amniotic fluid<70 d pregnancy period; nonfrozen amniotic fluid<70 d pregnancy period+glucose), nonfrozen allantoic fluid<70 d pregnancy period; and Whitten's medium. The percentages of embryos developing to hatched blastocyst stage were 66.6, 56.5, 57.4 and 63.9% respectively, for each of the four treatments. No differences were found between any two treatments (P<0.05). In Experiment 4 (n=231) the fluids were stored at -20 degrees C for 15 d. Whitten's medium was superior to amniotic or allantoic fluid<70 d pregnancy period in sustaining embryo development (P<0.05). In conclusion, these data indicate that nonfrozen bovine amniotic or allantoic fluid<70 d pregnancy period can support the development of murine embryos to the hatched blastocyst stage comparable to culture in Whitten's medium. Glucose supplementation of the amniotic fluid offered no advantage, and freezing of fluids had an adverse effect on in vitro embryo development.

To investigate incidence and causes of complete failed fertilization after intracytoplasmic sperm injection (ICSI) in a tertiary care facility. A total of 1,779 cycles between February 1994 and December 2003 were analyzed. Study parameters were female age, infertility diagnosis, ovarian stimulation protocol, estradiol level on day of hCG administration, number of follicles, number of oocytes retrieved, number of oocytes injected, and semen parameters. Complete failed fertilization occurred in 23 cycles (1.29%) involving a total of 85 oocytes injected. Infertility causes among patients with failed fertilization included unexplained (43.6%), male factor (26%), presence of more than one factor (17.4%), hysterectomy (4.4%), premature ovarian failure (4.3%), and advanced age (4.3%). In 12 cycles (52%), fewer than 5 follicles were present. In three (13%) cycles, no mature (MII) oocyte was available and in 61% (14/23) fewer than 3 MII oocytes were available for ICSI. Immotile sperm was use...