Effects of Exogenous Glutathione on the Quality of Chilled Bull Semen

Glutathione (GSH) at concentrations of 0.0 (control), 0.5, 1.0, 2.0 and 3.0 mM was added to chilled bull semen to determine its effects on the keeping quality of semen used for artificial insemination (AI). The semen was preserved with egg yolk citrate extender. All samples were stored at 4-8 degrees C for 5 days. Sperm motility and proportion of abnormal acrosome were assessed daily. Sperm motility was significantly (p < 0.01) higher in the semen treated with 0.5 mM glutathione than in untreated semen on each day. The optimum sperm motility (>or=50%) for AI was retained significantly (p < 0.01) for 3 days in 0.0, 0.5, 1.0 and 2.0 mM glutathione treated semen, whereas in 0.3 mM glutathione-treated semen, sperm motility was 46.8% for 3 days. Acrosomal damage was significantly (p < 0.01) reduced after addition of 0.5 mM GSH in the preserved semen. Bull semen can be preserved in chilled condition for 5 days with 0.5 mM GSH with sperm motility above 40% and 12% acrosome abnormality.     

MALDI‐MS Lipid Profiles of Oocytes Recovered by Ovum Pickup from Bos indicus and 1/2 indicus × taurus with High vs Low Oocyte Yields

The aim of the present study was to compare the lipid profile in oocytes of indicus and 1/2 indicus × taurus cows with high and low antral follicle count (AFC)/oocyte yields. After an OPU procedure (D0), antral follicles ≥3 mm were counted by ultrasonography (D4, 19, 34, 49, 64), and cows were assigned to groups with either high AFC (≥30 follicles; indicus, NH group; 1/2 indicus × taurus, AH group) or low AFC (≤15 antral follicles; indicus, NL group; 1/2 indicus × taurus, AL group). The lipid profiles of the oocytes were determined by MALDI‐MS. For GI, GII and GIII oocytes, the indicus samples tend to cluster separately from the 1/2 indicus × taurus samples. The lipid species [PC (P‐38:5) + H]⁺ and/or [PC (P‐36:2) + Na]⁺, [PC (38:2) + H]⁺, [PC (38:5) + Na]⁺ and [TAG (60:8) + NH⁴]⁺ were more abundant in indicus (NH and NL groups) than 1/2 indicus × taurus. The higher lipid content in the indicus oocytes likely reflects differences in the rate of lipid metabolism and may contribute to oocyte competence and embryo development.     

Purification of the ninth component of the bovine complement cascade.

Conditions for purification of the ninth component of bovine complement (C9) were established. The conditions for binding and elution from diethylaminoethyl cellulose and hydroxylapatite were different than for human C9. Serum albumin, a frequent contaminant of bovine C9 preparations, was removed by chromatography on reactive-red agarose. The calculated molecular weight of bovine C9 was 66,000, and reduction with 2-mercaptoethanol affected its migration on polyacrylamide gel electrophoresis. Some preparations of bovine C9 migrated as 2 bands when partially reduced, but extensively reduced preparations had a single band.     

Exogenous paraoxonase‐1 during oocyte maturation improves bovine embryo development in vitro

Paraoxonase‐1 (PON1) is an enzyme found in serum and follicular fluid that protects cell membrane and circulating lipids against oxidative damage. The aims of this study were to measure the direct effects of recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and to measure the carry‐over effects of PON1 on pre‐implantation embryo development in vitro. COCs were submitted to IVM with the addition of 0.0, 0.02, 0.04 and 0.08 mg ml^?1 of rPON1, corresponding to an average PON1 arylesterase enzyme activity of 2.2 ± 0.4, 15.5 ± 1.5, 30.2 ± 3.0 and 57.9 ± 5.0 U ml^?1, respectively. The results indicated that addition of rPON1 during IVM improved embryo development in a dose‐dependent manner as D7 embryo development was 22.2%, 29.4%, 32.2% and 37.0% for the treatment groups, respectively (p = 0.02). In conclusion, addition of PON1 enzyme during IVM exerted dose‐related positive effects on embryo development rates to blastocysts.     

The expression of histamine H4 receptor mRNA in the skin and other tissues of normal dogs

The histamine 4 (H₄) receptor was first cloned and characterized in 2000 using the human H₃ receptor DNA sequence. The H₄ receptor has been shown to participate in various aspects of inflammation, such as chemotaxis, upregulation of adhesion molecule expression and modulation of cytokine secretion. The primary goal of this study was to determine whether H₄ receptor mRNA is expressed in normal canine skin by performing an RT‐PCR. An additional goal was to determine the expression of this receptor in the colon, liver, spleen and kidney. Tissues were collected from five healthy, young‐adult pit bull dogs. Samples were immediately placed in RNAlater^® solution and stored at ?20°C until processed. The amplified products in all skin samples in addition to the colon, liver, spleen and kidney (variable expression) had the expected size of 400–500 bp. The sequenced amplicons matched the National Center for Biotechnology Information published sequence for the canine H₄ receptor. The study results showed that canine normal skin expresses the H₄ receptor mRNA. Further studies using immunohistochemistry should be conducted to demonstrate the expression of the H₄ receptor at the protein level and to localize the expression of this receptor in the skin.     

Lipid profiles of canine spermatozoa as revealed via matrix‐assisted laser desorption/ionization mass spectrometry

In this study, we investigated the ability of matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) to characterize the lipid contents of canine spermatozoa. For that, samples of pure semen were analysed. Indeed, quite comprehensive lipid coverage was observed, and the most abundant phospholipid ions detected were from four phosphatidylcholines, that is those of m/z 760.6; 782.6; 808.6; and 830.6 and one of m/z 725.6 from a sphingomyelin. In conclusion, MALDI‐MS was found to offer an easy, fast, accurate, and sensitive analytical method for lipid profiling in canine spermatozoa and could be used as a tool to select sires by assessing the relationship between sperm lipid profiles and variables such as age and breeding history as well as to study the effects of cryopreservation on lipid contents.     

Effect of oestrous synchrony between embryo donors and recipients,embryo quality and state on the pregnancy rate in beef cattle

This study aimed to evaluate the effect of oestrous synchrony between donors and recipients and the embryo quality on the pregnancy rate in beef cow recipients. The experiment was performed over two years at an embryo transfer (ET ) centre in Southern Brazil. Ninety Aberdeen Angus cows were subjected to superovulation (SOV ) protocols, resulting in the recovery of 1,048 transferable embryos. Eleven groups were formed with intervals of 6 hr, from ?30 to +30 hr, with respect to recipient versus donor oestrous detection. Evaluation of embryo quality was according to the IETS guidelines. The overall pregnancy rate was 52%. Effects related to donor and recipient oestrous synchronization on pregnancy rate were observed (p  =  .01), ranging from 36% to 50%. The embryo quality rate affected the pregnancy rate, where Grade I resulted in 57% and Grade III in 43% of pregnancy (p  <  .001). The embryonic state (frozen or fresh) showed no (p  >  .05) effect on pregnancy rate: 53% for fresh embryos and 44% for frozen embryos. The odds ratio for explanatory variables causing pregnancy indicated that Grade III embryos had 31% less chance of conception compared to Grade I. Thus, oestrous synchrony between donor and recipient, considering ±30 hr apart, can affect the pregnancy rate along with embryo quality.