Measuring Male-to-Male Differences in Fertility or Effects of Semen Treatments.

Fertility is a convenient but meaningless term unless the outcome measure is stipulated and accounts for dependence of male fertility on the female population. We describe outcome measures and detail the impacts of the physiological status of each female and her external environment, as well as management imposed by humans. We explain the dominant role of the female reproductive tract as a series of hurdles for sperm seeking an ovum. Each spermatozoon in an ejaculate is unique, although usually most are morphologically similar. Semen seemingly contains three subpopulations of sperm, based on fate within a female and role in hampering the success of the ultimate winning spermatozoon; we define these subpopulations. The numerical size of each subpopulation placed into a female determines the shape of the dose-response curve leading to possible live young. Heterospermic artificial insemination provides far greater sensitivity to detect differences, partly because the female environment is identical for each comparison.

Survival of honey bee (Hymenoptera: Apidae) spermatozoa incubated at room temperature from drones exposed to miticides.

We conducted research to examine the potential impacts ofcoumaphos, fluvalinate, and Apilife VAR (Thymol) on drone honey bee, Apis mellifera L. (Hymenoptera: Apidae), sperm viability over time. Drones were reared in colonies that had been treated with each miticide by using the dose recommended on the label. Drones from each miticide treatment were collected, and semen samples were pooled. The pooled samples from each treatment were subdivided and analyzed for periods of up to 6 wk. Random samples were taken from each treatment (n = 6 pools) over the 6-wk period. Sperm viability was measured using dual-fluorescent staining techniques. The exposure of drones to coumaphos during development and sexual maturation significantly reduced sperm viability for all 6 wk. Sperm viability significantly decreased from the initial sample to week 1 in control colonies, and a significant decrease in sperm viability was observed from week 5 to week 6 in all treatments and control. The potential impacts of these results on queen performance and failure are discussed.

Sperm morphology and preparation method affect bovine embryonic development.

This study was conducted to evaluate the effect of sperm separation methods of semen samples collected from bulls subjected to scrotal insulation on embryonic development after in vitro fertilization (IVF) and to determine whether IVF results would be affected by various heparin concentrations. Morphologically abnormal semen samples were obtained and cryopreserved from Holstein bulls following scrotal insulation for 48 hours. Standard protocols using the Percoll gradient (90%/45%) method and the swim-up method were used to separate spermatozoa fractions in experiment I. The pellet (A(p)) and the 45% layer (B(p)) were isolated from the Percoll separation, while for the swim-up separation, the supernatant (A(s)) and the interphase (B(s)) were isolated. The overall blastocyst rate for our laboratory control semen was 23.1 +/- 2.1% for Percoll separations (A(p) and B(p)) and 18.2 +/- 2.0% for swim-up (A(s) and B(s)) separations. This rate was higher (P <.01) than the rate observed for the semen from the bull that had the greatest response to scrotal insult 5 days prior to the insult, when it was 9.2 +/- 2.1% for the Percoll separation and 20.7 +/- 2.3% for the swim-up separation, while semen from 27 days after scrotal insulation (D +27) resulted in no blastocyst formation for the Percoll separation and a 4.2 +/- 2.1% rate for the swim-up separation. In experiment II, semen was sampled from the bulls that responded in the greatest and least degrees to scrotal insult 5 days before scrotal insulation (D -5) and on days 23 (D +23) and 34 (D +34) after scrotal insulation. These samples were exposed to IVF mediums with 3 different heparin concentrations (0.1, 1.0, and 10 microg/mL). There was a significant difference (P <.05) in developmental scores between the D -5 (1.08 +/- 0.08), D +23 (0.9 +/- 0.08), and D +34 (0.8 +/- 0.08) samples, but no differences were observed in blastocyst formation based on the number of cleaved embryos. Increasing the heparin concentration resulted in higher (P <.01) embryonic developmental scores. In conclusion, when semen samples with high percentages of abnormal spermatozoa are used for IVF, semen separation preparation methods affect results. Our results show that the separation methods used under these conditions were inadequate in their ability to provide potentially competent sperm for IVF. However, selecting appropriate sperm separation procedures could improve in the IVF embryonic development of semen from bulls used in artificial insemination. Also, an increase in the heparin concentration was able to partially overcome deficiencies, which suggests that morphologically abnormal spermatozoa undergo capacitation despite possible structural changes to the plasma membrane.

Effects of feeding endophyte-infected fescue seed on reproductive traits of male mice divergently selected for resistance or susceptibility to fescue toxicosis.

Our objective was to determine whether consumption of endophyte-infected fescue seed affected male reproduction differently in a mouse line previously selected for susceptibility (S) to fescue toxicosis than in a line previously selected for fescue toxicosis resistance (R). For 8 weeks following weaning, 48 males per line were provided diets containing 50% of either endophyte-infected (E+) or endophyte-free (E-) fescue seed. Each male was then paired with a female for 1 week, with litter size and weight recorded from subsequent births. Males were then killed, testes and seminal vesicles were weighed, cauda epididymal sperm were collected and testis cross-sections were fixed. The E+ diet reduced litter size by 0.5 in mates of S males but increased it by 1.0 in mates of R males (line by diet interaction P=0.05). Testis traits were not affected by diet or the line by diet interaction. Sperm integrity was adversely affected by the E+ diet (P<0.01) but did not differ significantly between lines, nor were line by diet interactions important. In earlier work, the E+ diet reduced long-term reproduction by a larger amount in S- than in R-line mated pairs. Because the E+ diet had similar effects on reproductive traits in R and S males in the current experiment, we infer that the differential impact previously reported acted primarily through traits expressed in females.

Examination of the binding ability of bovine spermatozoa to the zona pellucida as an indicator of fertility.

Despite the development of many new techniques, laboratory assays still do not predict male fertility accurately. To identify targets for laboratory assessment, we first need to determine which steps in fertilization are most often defective in subfertile males. We developed a competitive in vitro fertilization assay in which spermatozoa from 2 different males, stained with different lipophilic dyes, are incubated together with oocytes in a droplet. By exposing mixed spermatozoa to the same oocytes, this assay controls for many of the variables of in vitro fertilization and should allow identification of the most common faulty steps in fertilization. The relationship of zona-binding ability to fertility is controversial. Therefore, as a first step, we determined if zona pellucida-binding ability, measured by this competitive assay, was related to bovine spermatozoal fertility. Fertility data were collected from 2 groups of bulls by 2 means of evaluation, nonreturn to estrus rates postinsemination and competitive insemination. In the nonreturn to estrus study, semen samples from 15 bulls were effectively ranked by zona-binding ability, using pairwise competitive in vitro zona-binding assays (R(2) = 0.84). However, this ranking was not significantly correlated with nonreturn rates (r = -0.04). In the competitive insemination study, semen samples from 8 bulls were effectively ranked by pairwise comparison using the competitive zona-binding assay (R(2) = 0.67). Again, this ranking was not significantly correlated to the competitive insemination index calculated for these bulls (r = 0.29). In the third study, we tested 3 bulls to determine if in vivo zona binding, assessed by the number of accessory spermatozoa, was correlated with in vitro zona binding. The number of accessory spermatozoa on oocytes recovered from cows after mating was not correlated with in vitro competitive binding of the spermatozoa. In conclusion, in vitro competitive zona binding was not correlated with bovine fertility or binding of accessory spermatozoa to oocytes in vivo.