Genotoxicity in the Offspring of Rats Exposed to Contaminated and Acidified Experimentally Soils

The aim of this study was to evaluate the genotoxic and mutagenic potential of contaminated soil diluted in acidic solutions and not acidic, in the offspring of rats exposed during pregnancy and neonatal periods. To this end, a comet assay and micronucleus test were performed. Soil samples were solubilized in the following three solvents: distilled water (control group), acid solvent at pH 5.2, and acid solvent at pH 3.6. Soil and solvent were mixed in a rate of 1:2 in g/mL, and hydrofluoric acid was used in the acid solvents. In the comet assay, the tail length, percentage of DNA within the tail and tail moment was analyzed in the whole blood of the pups that were studied. The number of micronuclei found in the bone marrow cells was counted in the micronucleus test. In all parameters evaluated in the comet assay, the group exposed to the lowest pH value when associated with contaminated soil (p < 0.05) had the most damage. However, the micronucleus test showed differences between all exposed groups and the control group (p < 0.05). These results reaffirm the health risks related to the exposure to soil contaminants.     

Soil compaction caused by harvesting,skidding and wood processing in eucalyptus forests on coarse‐textured tropical soils

Eucalyptus forests play a major role in the world economy, providing raw materials for different purposes. In planted forests, harvest operations performed by heavy machinery may cause severe soil compaction. This study aimed to evaluate the impact of a full‐tree harvesting system in planted eucalyptus forests from Northeastern Brazil. Different soils were evaluated (two Hapludults and one Haplorthod) in two horizons (BA and Bt). We tested different equipment, namely feller buncher, skidder (with traffic intensities of 3, 6, 12 and 16 passes), flail (at different ground‐contact points), grapple saw and loader. The soil physical attributes reflected not only the impact of equipment traffic but also the intrinsic differences between the soils. Bulk density (ranging from 1.36 to 1.80 t m^?3 after trafficking) related well to soil class and horizon. Precompression stress (ranging from 203 to 430 kPa) and degree of compaction (76%–94%) following trafficking were well correlated, while increase in bulk density (reaching a maximum of 20%) related more strongly to soil moisture. A contingency table was constructed with the number of compacted samples and further examined by correspondence analysis. Compaction varied according to soil, horizon and equipment, indicating that machine–soil interactions are very specific and demand detailed research under different conditions. The Haplorthod experienced the greatest amount of compaction, whereas the Hapludult‐2 was more resistant (60% and 25% of compacted samples, respectively). The grapple saw and the skidder at higher traffic intensities (12 and 16 passes) exerted the highest mechanical impacts (81% and 67% of compacted samples, respectively).     

Effect of melatonin on DNA damage of bovine cumulus cells during in vitro maturation (IVM) and on in vitro embryo development

The effect of melatonin during in vitro maturation (IVM) on DNA damage of cumulus cells (CCs) from bovine cumulus-oocyte complexes (COCs) and embryo development was evaluated. COCs from abattoir ovaries were cultured in maturation medium (MM) with 0.5μg/ml FSH and 5.0μg/ml LH (FSH-LH); 10(-9)M melatonin (MEL) or FSH-LH+MEL (FSH-LH-MEL). After 24h of in vitro maturation, the CCs surrounding the oocyte were subjected to DNA analysis by Comet assay. After in vitro fertilization and in vitro embryo culture, the embryo development rates were evaluated on day 2 post insemination (cleavage) and days 7-8 (blastocyst). The percentage of CCs with no DNA damage was significantly superior in MEL group (37.6±2.4) than in FSH-LH-MEL (28.0±2.4) and FSH-LH (17.8±2.41) groups. Cleavage and blastocysts rates were similar among groups. Melatonin during IVM protects the CCs from DNA damage but this effect did not influence embryo development in vitro.     

Pre-parturition staphylococcal mastitis in primiparous replacement goats: persistence over lactation and sources of infection

This investigation reported for the first time the occurrence of intramammary infections caused by Staphylococcus in primiparous replacement goats before parturition and the persistence of clinical Staphylococcus aureus infection during the lactation period. Subclinical infections, mainly caused by coagulase negative staphylococci (CoNS), did not persist during lactation. Genotyping analysis indicated that environment seems to play a moderate role as source of intramammary infections to goats before parturition, but causative agents of mastitis in lactating animals are not genotypically related to environmental staphylococci. The occurrence and persistence of intramammary infections in replacement goats demonstrate the need to consider those animals as potential sources of infections in dairy goat herds.     

Comparison of Conventional Freezing and Vitrification with Dimethylformamide and Ethylene Glycol for Cryopreservation of Ovine Embryos

The aim of this work was to evaluate the efficiency of the cryoprotectants dimethylformamide and ethylene glycol for cryopreservation of ovine embryos using vitrification and conventional freezing. The recovered embryos were distributed randomly in three treatment groups: Gr. 1: conventional freezing (n = 44), Gr. 2: vitrification with ethylene glycol (n = 39) and Gr. 3: vitrification with dimethylformamide (n = 38). Quality of fresh embryos in control group as well as of frozen and vitrified embryos was examined by three methodologies: staining with propidium iodide and Hoechst 33258 and evaluation under fluorescent microscopy, evaluation of re‐expansion and hatching rates after culture, and determination of apoptotic index with TUNEL technique. It was established that re‐expansion rate in all treatment groups was similar. In the same time, hatching rates were higher in Gr. 1 (40.5%) and Gr. 2 (35.3%) in comparison with Gr. 3 (15.5%, p < 0.05). The number of dead cells in vitrified embryos of Gr. 2 and Gr. 3 was higher (42.6 ± 26.2 and 63.2 ± 34.65, respectively) in comparison with Gr. 1 (conventional freezing, 10.1 ± 8.5, p < 0.05). Embryos vitrified with dimethylformamide included the same quality of apoptotic cells that Gr. 1 (conventional freezing) and fresh embryos. In conclusion, the dimethylformamide and ethylene glycol used as cryoprotectant to vitrify ovine embryos, in the concentrations and exposition time tested in this work, were not as efficient as the conventional freezing for cryopreservation of ovine embryos Thus, the conventional freezing with ethylene glycol was the most efficient method to cryopreserve ovine embryos in comparison with vitrification.     

Viability of OPS Vitrified Sheep Embryos After Direct Transfer

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.