The relationship between sperm nuclear chromatin structure and fertility was evaluated in two groups of Holstein bulls: Group 1, 49 mature bulls, and Group 2, 18 young bulls. Fertility ratings had been estimated for Group 1 and nonreturn rates were known for Group 2. Semen samples were measured by the sperm chromatin structure assay (SCSA): sperm were treated to induce partial in situ DNA denaturation, stained with acridine orange, and evaluated by flow cytometry. Acridine orange intercalated into double-stranded DNA emits green fluorescence upon excitation with 488 nm light, and red fluorescence when associated with single-stranded DNA. An index of DNA denaturation per cell is provided by alpha-t [alpha t = red/(red + green) fluorescence]. The standard deviation (SD alpha t), coefficient of variation (CV alpha t) and proportion of cells outside the main population (COMP alpha t) of the alpha t distribution quantify the extent of denaturation for a sample. Intraclass correlations of the alpha t values were high (greater than or equal to 0.70), based on four collections obtained over several years from Group 1 bulls. Negative correlations were obtained between fertility ratings and both SD alpha t (-0.58, p less than 0.01) and COMP alpha t (-0.40, p less than 0.01) in Group 1, and between nonreturn rates and both SD alpha t (-0.65, p less than 0.01) and COMP alpha t (-0.53, p less than 0.05) in Group 2. These data suggest that the SCSA will be of value for identification of low fertility sires and poor quality semen samples.
In three experiments, mice from lines selected for resistance (R) or susceptibility (S) to growth depression from endophyte-infected fescue seed in the diet were fed diets containing infected (E+) or non-infected (E-) seed. Activities of liver enzymes known to participate in oxidation, reduction, or hydrolysis or in conjugation of xenobiotics were measured in these mice. In all experiments, E+ caused greater reduction in initial ADG of S than of R mice. In Exp. 1, liver cytochromes P450 and b5 activities were not affected by line, diet, or their interaction. These enzymes were not evaluated in subsequent experiments. In all experiments, glutathione-S-transferase (GST) and uridine diphosphate glucuronosyltransferase (GRT) activities differed between lines. Resistant mice had significantly higher GST activity on both diets in Exp. 1, on E- in Exp. 2, and on E+ in Exp. 3. Resistant mice had higher GRT activities on E+ in Exp. 1, on E- in Exp. 2, but after 4 wk on either diet in Exp. 3. Before test diets were imposed in Exp. 3, GST and GRT activities were higher in R-line mice. Divergent selection created lines that differed in response to tall fescue in the diet. Postweaning growth of resistant mice was less severely depressed by E+, although susceptible mice later expressed compensatory gain. Activities of two detoxification enzymes generally were higher in livers from R-line mice, suggesting a biochemical mechanism for the difference. Using such traits, it may be possible to select ruminants for resistance to fescue toxicosis.
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