Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Evaluation of nitrate and ammonium as sources of NO and N2O emissions from black earth soils (Haplic Chernozem) based on 15N field experiments

Nitrous oxide (N₂O) and nitric oxide (NO) released from soil is a concern since it can act as a potential atmospheric pollutant and it represents a loss of N from the soil. These gases are present in the atmosphere in trace amounts and are important to atmospheric chemistry and earth's radiative balance. Nitric oxide (NO) does not directly contribute to the greenhouse effect, but it contributes to climate forcing through its role in photochemistry of hydroxyl radicals and ozone and plays a key role in air quality issues. Nitrification and denitrification have been identified as major controlling microbial processes in soils responsible for the formation of NO and N₂O. To elucidate the contribution of both processes to the release of NO and N₂O from loess-black earth soils under field conditions—i.e. to evaluate nitrate and ammonium as sources of NO and N₂O emission—two field experiments with either [¹⁵N] nitrate (NO₃^?) or [¹⁵N] ammonium (NH₄⁺) labelling have been conducted at two sites differing in soil organic matter content (high and normal SOM). [¹⁵N] nitrate treatments revealed that denitrification of NO₃^? represents the main pathway of soil N₂O release. On average 76% and 54% of N₂O was emitted during denitrification from soils with high and normal SOM content, respectively. Contrarily, denitrification contributed on average only 17% and 12% of released NO from soil with high and normal SOM content, respectively. The [¹⁵N]ammonium treatments revealed that nitrification of NH₄⁺ is the major process responsible for soil NO emission. SOM content of the loess-black earth soil significantly influenced NO and N₂O emission. The soil with the higher SOM content showed lower NO emission but drastically increased N₂O emission after nitrate fertilisation. In particular the soil with high SOM content exhibited a high sorption capacity for ammonium ions which led to unexpected results after fertilisation with [¹⁵N]ammonium. To explain this results a revised concept containing three different interacting soil ammonium pools have been hypothesised.     

Synthesis of macroporous minerals with highly ordered three-dimensional arrays of spheroidal voids

Titania, zirconia, and alumina samples with periodic three-dimensional arrays of macropores were synthesized from the corresponding metal alkoxides, using latex spheres as templates. In a fast, single-step reaction, the monomeric alkoxide precursors permeate the array of bulk polystyrene spheres and condense in air at room temperature. Close packed, open-pore structures with 320- to 360-nanometer voids are obtained after calcination of the organic component at 575 degreesC. The examples presented demonstrate the compositional diversity possible with this technique. The resulting highly structured ceramics could have applications in areas ranging from quantum electronics to photocatalysis to battery materials.     

Measurement and simulation of herbicide transport in macroporous soils

Lysimeter experiments were carried out to study pesticide transport through macroporous soils. In order to differentiate between the effects of soil structure and chemical behaviour, the leaching experiments were conducted using disturbed and undisturbed soil samples. Two herbicides with different sorption behaviours, and bromide as tracer were applied. The results were used to validate a dynamic simulation model which considers bypass flow in macropores. The simulation results show that the model is able to reproduce the soil suction within the soil as well as the spatial distribution of bromide and the herbicides. The continuity of the macropores is most important for the efficiency of bypass flow. The results indicate that cultivation practices like ploughing significantly influence the temporal and spatial distribution of the macropores. © 1998 SCI.     

Role of nitrite and nitric oxide in the processes of nitrification and denitrification in soil: Results from N tracer experiments

Recent research has proven soil nitrite to be a key element in understanding N-gas production (NO, N₂O, N₂) in soils. NO is widely accepted to be an obligatory intermediate of N₂O formation in the denitrification pathway. However, studies with native soils could not confirm NO as a N₂O precursor, and field experiments mainly revealed ammonium nitrification as the source of NO. The hypothesis was constructed, that the limited diffusion of NO in soil is the reason for this contradiction. To test this diffusion limitation hypothesis and to verify nitrite and NO as free intermediates in native soils we conducted through-flow (He/O₂ atmosphere) ¹⁵N tracer experiments using black earth soil in an experimental set up free of diffusion limitation. All of the three relevant inorganic N soil pools (ammonium, nitrite, nitrate) were ¹⁵N labelled in separate incubation experiments lasting 81 h based on the kinetic isotope method. During the experiments the partial pressure of O₂ was decreased in four steps from 20% to about 0%. The net NO emission increased up to 3.7 μg N kg⁻¹ h⁻¹ with decreasing O₂ partial pressure. Due to the special experimental set up with little to no obstructions of gas diffusion, only very low N₂O emission could be observed. As expected the content of the substrates ammonium, nitrate and nitrite remained almost constant over the incubation time. The ¹⁵N abundance of nitrite revealed high turnover rates. The contribution of nitrification of ammonium to the total nitrite production was approx. 88% under strong aerobic soil conditions but quickly decreased to zero with declining O₂ partial pressure. It is remarkable that already under the high partial pressure of 20% O₂ 12 % of nitrite is generated by nitrate denitrification, and under strict anaerobic conditions it increases to 100%. Nitrite is present in two separate endogenous pools at least, each one fed by the nitrification of ammonium or the denitrification of nitrate. The experiments clearly revealed that nitrite is almost 100% the direct precursor of NO formation under anaerobic as well as aerobic conditions. Emitted N₂O only originated to about 100% from NO under strict anaerobic conditions (0-0.2% O₂), providing evidence that NO is a free intermediate of N₂O formation by denitrification. To the best of our knowledge this is the first time that NO has been detected in a native soil as a free intermediate product of N₂O formation at denitrification. These results clearly verify the “diffusion limitation” hypothesis.     

Long‐term Effects of Pyrethrin and Cyfluthrin,a Type II Synthetic Pyrethroid,Insecticide Applications on Bull Reproductive Parameters

The objectives of this study were to determine effects of cyfluthrin and pyrethrin spray products, used in combination with cyfluthrin topical and ear tag applications, on bull reproductive parameters over 18 weeks. Angus or Angus x Simmental bulls were randomly assigned to one of three treatment groups: (i) no exposure to pyrethrins/cyfluthrin (CONT; n = 10), (ii) cyfluthrin ear tag and topical applications (ET; n = 10), or (iii) cyfluthrin ear tag, topical, premise spray and pyrethrin fog spray applications (ET+S; n = 8). Bull body weight was measured every 3 week, and body condition score and scrotal circumference were recorded on weeks 0, 9 and 18. Semen and serum were collected every 3 weeks for sperm evaluation and testosterone measurement, respectively. There was a treatment × week interaction (p < 0.01) for sperm with primary defects; bulls in CONT group had a greater (p = 0.01) percentage of sperm with primary defects than bulls treated with insecticides at week 18. Overall and progressive sperm motility, normal sperm morphology, secondary sperm defects and serum testosterone concentrations changed (p < 0.01) over time in all bulls; however, treatment did not affect (p ≥ 0.13) any of these parameters. There were also no treatment effects (p ≥ 0.08) on bull body weight, body condition score or scrotal circumference. The use of pyrethrin‐ and cyfluthrin‐based insecticides, regardless of application, did not negatively affect reproductive parameters in beef bulls when administered over 18 weeks.     

A cross-sectional survey on wounds in horses in New Zealand

AIM: To determine the frequency and type of skin wounds encountered by New Zealand veterinarians in their equine patients, the duration and estimated costs of treatment as well as the expected outcomes for these wounds.

METHODS: An online survey was sent to all veterinarians registered with the New Zealand Equine Veterinary Association. The survey comprised questions on the location and experience of respondents, the number of wound-related cases in relation to the total equine caseload, the type and anatomical location of wounds treated, the frequency, duration and costs of treatments, the outcome of wound treatment and an estimate of the most common causes of death or euthanasia in their equine patients.

RESULTS: The survey response rate was 110/262 (41.9%). The median number of equine cases seen by respondents was 20 (interquartile range (IQR) 6–60) per month; of these, five (IQR 2–10) were wound related. Wounds ranked third after lameness and respiratory disease for the relative frequency with which respondents encountered them. Of 102 respondents 59 (58%) reported that their clients frequently treated wounds incurred by their horse without consulting a veterinarian. Wounds on the distal limb, whether involving only the skin or also deeper structures, were reported by 86/101 (85%) respondents as the most frequently encountered. Wounds in this location also incurred the longest treatment period and were the most prone to develop complications. Finally, wounds ranked second, after colic, as the most common cause of death/euthanasia in the veterinary respondents’ equine patients.

CONCLUSIONS: The data obtained via the survey indicate that skin wounds, particularly on the distal limb, are a common occurrence in horses in New Zealand and, when they involve structures underlying the skin, are costly and time-consuming to manage and may lead to decreased performance, retirement or euthanasia. Consequently, we recommend that more effort be devoted to the education of equine veterinarians and owners, and that appropriate research funds be allocated to help improve patient outcomes.     

Use of a Low Dose of Equine Purified FSH to Induce Multiple Ovulations in Mares

The effects of a low dose of equine purified FSH (eFSH) on incidence of multiple ovulations and embryo recovery rate in mares were studied. During the physiological breeding season in Brazil (19°45′45′S), 14 Mangalarga Marchador donor mares were used in a crossover study and another 25 mares of the same breed, between 3 years and 12 years of age were used as recipients for the embryo transfers. Donors were monitored during two consecutive oestrus cycles, an untreated control cycle followed by a treated cycle, when eFSH was administered. In both cycles, after an embryo collection attempt on day 8 post‐ovulation all mares received 7.5 mg dinoprost and had their two largest follicles tracked daily by ultrasonography until the period of ovulation. Mares were inseminated every 48 h with extended fresh semen from a single stallion after the identification of a 35‐mm follicle until the period of ovulation. Ovulations were induced by intravenous administration of 2.500 IU of human chorionic gonadotropin, upon detection of a 35‐ to 40‐mm follicle. In the treated cycle, 5 mg eFSH was given intramuscularly once a day, from day 8 post previous ovulation until at least one follicle reached 35 mm in diameter. Embryo flushes were performed on day 8 of dioestrus (day 0 = ovulation). Treatment with eFSH resulted in higher (p < 0.05) ovulation rate and incidence of multiple ovulations compared to the control (1.6 vs 1.0 and 50% vs 0%, respectively – one mare had triple ovulation). However, embryo recovery rates in the control and treated cycles were similar (0.8 and 1.0, respectively; p > 0.05). Pregnancy rates in the recipient mares following embryo transfer were similar for the control and eFSH cycles (11/11 and 10/14, respectively). Additional studies are necessary in order to develop a low‐dose protocol for the use of eFSH that brings a more consistent contribution to the efficiency of commercial equine embryo transfer programs.