Perturbations in the uterine luminal fluid composition are detrimental to pregnancy establishment in cattle

Background: A major,unresolved issue is how the uterine microenvironment determines pregnancy success in cattle.Before implantation,conceptus development depends on the uterine secretome(i.e.,histotroph).Despite its pivotal role,little is known about the dynamics of histotroph synthesis and changes in composition throughout the early diestrus and the relevance to pregnancy establishment.We hypothesize that disturbances on histotroph composition affect the establishment of pregnancy.Aim was to disturb histotroph composition at early diestrus and verify the effects on:(Exp.1) timing to restore its composition; and(Exp.2) pregnancy rate after multiple-embryo transfer.Estrous cycle of multiparous Nelore cows were synchronized and estrus was considered d 0(D0) of the experiments.Disturbance was through flushing each uterine horn with 30 m L of DMPBS and collecting the resulting uterine luminal flushing(ULF) on D1; D4; D7; D1 + D4 + D7.Control group remained not-collected.In Exp.1,ULF was collected on D7.5 from all animals and used for quantification of total protein concentration and abundance of albumin.In Exp.2,three in vitro-produced embryos were transferred to the uterine horn ipsilateral to the ovary containing the CL on D7.5 and pregnancy was checked on D25 by ultrasound.Results: In Exp.1,ULF collection on D4 or D7 increased(1.5-to 2.2-folds) the total protein concentration and albumin abundance.ULF collection on D1 did not alter(P 0.10) these endpoints.In Exp.2,ULF collected on D4 or D7 decreased pregnancy rates to approximately half of that measured in the remaining groups.Conclusions: Subtle perturbations imposed to the native intrauterine milieu,such as those caused by a single,low-volume collection of ULF,profoundly disturbs intrauterine composition and pregnancy success.At least 4 d were necessary for the uterus to recover its composition and the functional capacity to carry post-implantation gestation.     

Ibuprofen Genotoxicity in Aquatic Environment: An Experimental Model Using Oreochromis niloticus

Medicines and their metabolites have been found as water contaminants at very low concentrations; moreover, there is no extensive toxicological data to determine the risks associated with their occurrence in water resources. The ibuprofen genotoxicity potential to the Oreochromis niloticus fish (Tilapia), due to nanograms per liter exposure, was evaluated using the micronucleus test. Acute (48 h) and sub-chronic assays (10 days) were carried out at 300 ng/L ibuprofen aquatic concentration comparing with the negative control group (without treatment), with eight animals per group. The results were assessed from the average of triplicate analyses. The micronucleus frequency in peripheral blood of fish was determined using a sample size of 3,000 erythrocytes per animal. Significance was defined using t test (p????0.05). The bioassay results showed a statistically significant increase in the frequency of micronuclei for both exposure times in comparison to the negative control. The micronucleus frequency observed for the sub-chronic tests was higher than the one identified in the acute assays. The observed ibuprofen genotoxic effects demonstrated an aquatic environmental risk of this pharmaceutical, which occurs for the used fish experimental model in lower concentration than previously described for other aquatic organisms.     

Genotoxic Effects in Erythrocytes of Oreochromis niloticus Exposed to Nanograms-per-Liter Concentration of 17??-Estradiol (E2): An Assessment Using Micronucleus Test and Comet Assay

Pharmacologically active substances used in the treatment of human and animal illnesses may usually enter the aquatic environment via effluents from sewage treatment plants, as they are not completely biodegraded or removed during waste water treatment. 17??-Estradiol genotoxicity was evaluated in Oreochromis niloticus (family Cichlidae) using micronucleus test, other nuclear abnormalities assessment, and the comet assay with erythrocytes. Fish were exposed to aqueous systems contaminated with 6 ng/L 17??-estradiol for three periods: 24 h, 48 h, and 10 days. The results showed that 17??-estradiol has genotoxic potential in different periods, since significant differences (P?=?0.036) were observed in the micronucleus frequencies of the 10-day exposure groups in relation to the control group. Also, the same was observed when comparing the nuclear abnormality frequencies (P?=?0.018) of the 24-h exposure group with the negative control group, and when using comet assay (P?<?0.001) for 48-h evaluations. The tested concentration of the 17??-estradiol gave rise to mutagenic and genotoxic effects on the blood cells of O. niloticus, therefore the substance being considered a clastogenic chemical contaminant on both acute and chronic exposures. The assessment using a combination of micronucleus test, nuclear abnormalities, and comet assays proved to be suitable and useful in the genotoxicity testing of 17??-estradiol at nanograms per liter.     

Environmental Degradation at a Public Park in Southern Brazil as Revealed Through a Genotoxicity Test (MN) on Peripheral Blood Cells from Poecilia vivipara (Teleostei)

The effects of anthropogenic activities on water, environment, and consequently quality of life can be evaluated using genetic, biochemical, and microbiological parameters. Regarding genetic parameters, the micronucleus test is a fast, efficient, inexpensive method for detecting alterations in genetic material induced by a variety of genotoxic agents. In the present study, blood cells from Poecilia vivipara from the Belém River in the city of Curitiba, Paraná, Brazil were evaluated for genotoxic effects stemming from human-produced pollution, as expressed by the micronucleus. The water in the river was evaluated with regard to physiochemical and microbiological parameters as well as for heavy metals. The analysis revealed the presence of copper, zinc, and nickel, with high concentrations of copper. The micronucleus analysis revealed significant differences in relation to the groups (study and control), suggesting a positive relation between the water quality of the Belém River and micronucleus expression as a result of the pollution to which this river is subjected.     

Effects of cysteine supplementation on the quality of cryopreserved sperm of South American silver catfish

The cryopreservation promotes cellular damage that could compromise sperm quality in terms of motility and fertility rates, which may be caused by oxidative stress. Thus, the aim of this study was to assess the effects of cysteine addition on post‐thaw sperm quality, DNA damage and indices of oxidative stress of the South American silver catfish (Rhamdia quelen) sperm, compared with the cryoprotectant solution without cysteine addition. Sperm collected from five males were cryopreserved in cryoprotectant solution (fructose 50 g/L, powdered milk 50 g/L and methanol 100 ml/L) containing different cysteine concentrations (0, 2.5, 5, 10 and 20 mM). After thawing, the following were measured: sperm motility, morphology, sperm viability, DNA damage, lipid peroxidation, concentration of carbonyl and sulfhydryl groups and the activity of SOD, CAT, GST and GPx enzymes. The lowest sperm motility was determined for semen cryopreserved with addition of 20 mM of cysteine. The control group had the lowest DNA damage and lipid peroxidation. The findings of this study show that cysteine addition had no positive effect on evaluated parameters. Therefore, the concentrations tested are not recommended for the supplementation of cryoprotectant solution for semen of R. quelen.     

Response of Arbuscular Mycorrhizal Fungi in Different Soil Tillage Systems to Long‐Term Swine Slurry Application

Swine slurry is a common agricultural fertilizer in many countries. However, its long‐term use in large amounts can cause excess nutrient accumulation, alter soil compounds, and potentially influence critical microbial populations such as arbuscular mycorrhizal fungi, which have important roles in plant nutrition and soil sustainability. This work determined if arbuscular mycorrhizal status, external mycelium, and glomalin‐related soil protein content were affected by long‐term swine slurry application to different soil tillage systems. The experiment was conducted on a clayey oxisol, in southern Brazil. Swine slurry (0, 30, 60, 90, and 120 m³ ha⁻¹ y⁻¹) was applied for 15 years to conventional tillage and no tillage soil prior to the summer (soybean or maize) and winter (wheat or oats) crop seasons. Swine slurry decreased mycorrhizal root colonization, spore number, and total external mycelium. Swine slurry increased active external mycelium and both easily extractable and total glomalin‐related soil protein. No‐tillage soil had more glomalin‐related soil protein than conventional tillage soil. The most significant response variables were root colonization, easily extractable glomalin‐related soil protein, and total external arbuscular mycorrhizal mycelia. Long‐term application of swine slurry in this environment, even at high rates, did not adversely affect crop yield but did influence arbuscular mycorrhizae fungi and their products in the soil environment. Benefits of swine slurry application for crop nutrition must be weighed against potential adverse consequences for the size, activity, and benefits of the mycorrhizal community to subsequent annual crops. Copyright © 2014 John Wiley & Sons, Ltd.