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.     

HPLC separation and determination of 12 cholesterol oxidation products in fish: comparative study of RI, UV, and APCI-MS detectors

A simple, fast, and sensitive method for the extraction through direct saponification, separation, quantification, and identification of 12 cholesterol oxidation products (COPs) and cholesterol in a single isocratic, normal-phase, high-performance liquid chromatography (HPLC) was developed. Three detectors were compared for determination of COPs and cholesterol in fish samples: refractive index (RI), ultraviolet (UV), and atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The results did not show significant differences (p > 0.05) between the concentration of the cholesterol oxides and cholesterol obtained with these detectors. The present study demonstrated the presence of 19-hydroxycholesterol, 22"R"-hydroxycholesterol, 22"S"-hydroxycholesterol, 24"S"-hydroxycholesterol, and 25"R"-hydroxycholesterol for the first time in fish samples.     

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.     

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.     

Electrospray ionization mass spectrometry fingerprinting of Brazilian artisan cachaça aged in different wood casks

We have investigated the capability of direct infusion electrospray ionization mass spectrometry in the negative ion mode, ESI(-)-MS, to differentiate representative samples of artisan cacha?a, a Brazilian sugar cane distillate of large production, aged in four different types of wood casks: amburana (Amburana cearensis), jequitibA (Cariniana estrellensis), balm (Myroxylon peruiferum), and oak (Quercus rubra). The ESI(-)-MS were found to be very characteristic, showing sets of diagnostic ions for each of the four types of samples: amburana (m/z 271, 313, 377), jequitibA (m/z 143, 171, 255), balm (m/z 137, 269, 283, 297), and oak (m/z 197, 301, 307). Furthermore, principal component (PCA) and hierarchical cluster analysis (HCA), applied to the ESI(-)-MS data, divided these samples into four definite categories. The influence of the aging time on the ESI(-)-MS fingerprints of the cacha?a samples stored in oak casks was also established. An inversion in the relative intensity of the diagnostic ions of m/z 307 and 301 is detected in the ESI(-)-MS as the aging time increased from 1 to 2 years. The chemical structures of the major cacha?a components were proposed on the basis of the following: (a) the comparison of the ESI(-)-MS/MS of the diagnostic anions with those of the authentic anions or (b) the interpretation of the fragmentation patterns of the previously unknown diagnostic anions. Hence, direct infusion ESI(-)-MS allows not only a rapid, simple, and accurate way to distinguish among cacha?a samples stored in different wood casks but also monitoring changes in their chemical composition according to the aging time.     

Radiographic assessment of pulmonary fluid clearance and lung aeration in newborn calves delivered by elective Caesarean section

The aim of this study was to develop a radiographic standard for the assessment of pulmonary fluid clearance and lung aeration in newborn calves. Caesarean‐delivered mature calves (n = 9) underwent lung assessment by thoracic radiography as well as arterial and venous blood gas analysis within the first 30 min, 1, 2, 3, 6, 12 and 24 hr after birth. The results indicated that newborn calves delivered by elective Caesarean section suffered from a physiological combined respiratory and metabolic acidosis with the dominance of respiratory acidosis, and an improvement in these conditions was recorded within 24 hr after birth. Concerning the radiographic results, clear lung fields, improvement in lung expansion, air content of the lung and absence of lung opacification occurred within 24 hr of birth. Furthermore, the ventral lung quadrant showed an improvement in radiographic opacification and lung expansion earlier than the dorsal lung regions. The findings of this study support the potential role of thoracic radiography in the assessment of pulmonary fluid clearance and lung aeration in newborn calves.     

Arabica and robusta coffees: identification of major polar compounds and quantification of blends by direct-infusion electrospray ionization-mass spectrometry

Considering that illegal admixture of robusta coffee into high-quality arabica coffee is an important task in coffee analysis, we evaluated the use of direct-infusion electrospray ionization-mass spectrometry (ESI-MS) data combined with the partial least-squares (PLS) multivariate calibration technique as a fast way to detect and quantify arabica coffee adulterations by robusta coffee. A total of 16 PLS models were built using ESI± quadrupole time-of-flight (QTOF) and ESI± Fourier transform ion cyclotron resonance (FT-ICR) MS data from hot aqueous extracts of certified coffee samples. The model using the 30 more abundant ions detected by ES+ FT-ICR MS produced the most accurate coffee blend percentage prediction, and thus, it was later successfully employed to predict the blend composition of commercial robusta and arabica coffee. In addition, ESI± FT-ICR MS analysis allowed for the identification of 22 compounds in the arabica coffee and 20 compounds in the robusta coffee, mostly phenolics.