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.     

Morphological variability of Atlantic salmon Salmo salar and brown trout Salmo trutta in different river environments

Abstract – The morphologies of sympatric juvenile Atlantic salmon Salmo salar parr and brown trout Salmo trutta parr were compared between Irish rivers with contrasting hydraulic environments – a high‐gradient and a low‐gradient tributary from the River Barrow catchment, south‐east Ireland and a high‐gradient river from the Burrishoole catchment, west of Ireland. The two catchments differ markedly in mean annual precipitation (849.0 mm year^?1 and 1370.3 mm year^?1 for the Barrow and Burrishoole catchments, respectively). Parr of both species demonstrated morphological variation between and within catchments. Changes in metrics such as pectoral fin length, body depth and body length between and within catchments suggest that hydraulic forces were a major determinant of morphological variation. Both species from the Burrishoole catchment had relatively larger pectoral fins, longer heads, larger eyes and shallower bodies than conspecifics from the Barrow catchment. In rivers subject to frequent rainfall‐driven high discharges, such as those in the Burrishoole catchment, a more fusiform body and head shape coupled with larger pectoral fins may reduce the energetic cost of maintaining position in the water column, as well as increase stability and manoeuvrability. The larger eyes in both parr species in the Burrishoole catchment could further be a response to the lower visibility of the more turbid and peat‐stained waters or to the reduced prey availability. The results of this study demonstrate that local adaptation to flow conditions can yield morphologically distinct populations and that multiple species can exhibit parallel phenotypic responses to changing environmental conditions.     

Recent advances in external skeletal fixation

While the use of external skeletal fixation was once associated with substantial postoperative morbidity, clinical and experimental studies have led to technological advances and modifications in application techniques that have greatly improved the results obtained with this treatment modality. The past decade saw numerous advances in external skeletal fixator implants, components and instrumentation, including improvements in fixation pin design, and the development of new linear external skeletal fixation systems and economical circular external skeletal fixation systems specifically engineered for use in dogs and cats. In addition, a greater understanding of fixator biomechanics and the pathobiology of the bone-fixation pin interface have improved fixator application practices. This article reviews many of the more significant recent advances in external skeletal fixation.     

Artificial Insemination of Gilts with 1.5 Billion Sperms Stored in Different Periods Associated with Different Pre-ovulatory Intervals

This study evaluated the reproductive performance of gilts inseminated at three intervals before ovulation (0-12, 13-23, 24-30 h) with sperm doses (SD) stored for 0-48 and 96-120 h. A total of 218 PIC Camborough 22 gilts were inseminated once with SD of 1.5 x 10(9) sperms. Pregnant gilts (n = 166) were slaughtered 30.8 +/- 3.7 days after artificial insemination. The number of corpora lutea (CL) and total embryos (TE) was counted. Pregnancy rates (PR) were analysed by chi-square test. TE and embryonic survival (ES), obtained as the ratio between viable embryos and CL, were analysed by GLM procedure (SAS) and mean values were compared by Tukey's test. Pregnancy rate was similar among artificial insemination-ovulation (AIOV) intervals when semen was stored for 0-48 h. However, the lowest PR was observed in the 24-30 h AIOV interval with storage time (ST) of 96-120 h (p < 0.05). There was a significant effect of the interaction between ST and AIOV (p < 0.05) on TE and ES variables. Total embryos and ES did not differ (p > 0.05) among AIOV intervals in ST of 0-48 h. However, gilts inseminated at 24-30 h AIOV interval with ST of 96-120 h showed a reduction of 6.7 embryos (p < 0.05) compared with gilts in the same interval inseminated with semen stored for 0-48 h. ES for the 24-30 h AIOV interval and ST of 96-120 h was lower than that observed in the other groups (p < 0.05).     

Oxidative responses in ferret macrophages

Although the basic function of T and B lymphocytes in ferrets has been known for some time, the function of mononuclear phagocytes has not been described in this species. The present study has characterised basic oxidative responses in ferret macrophages, and has investigated the effects of endogenous and exogenous modulators of macrophage function on oxidative capacity in vitro. Macrophages derived from the blood or lungs of ferrets were shown capable of generating the reactive oxygen intermediate (ROI) molecules superoxide and hydrogen peroxide, and secreting a lysosomal enzyme (acid phosphatase), in response to appropriate stimuli. A T cell supernatant (derived from mitogen-stimulated peripheral blood lymphocytes) was able to activate both blood- and lung-derived macrophages for enhanced ROI production, while specific ROI inhibitors (superoxide dismutase and catalase) were able to partially ablate ROI activity. The accumulation of nitrite in culture supernatants, as an indicator for the production of reactive nitrogen intermediates, could not be demonstrated by ferret macrophages derived from either tissue source. In contrast to the enhancing effects of TCS on the oxidative function of blood-derived macrophages, exposure to bacterial LPS caused marked suppression of ROI and lysosomal enzyme production by these cells. Finally, the generation of superoxide anion, following phagocytosis of live or heat-killed Mycobacterium bovis or zymosan, indicated that ROI production in response to phagocytic stimulation was relatively weak in ferret blood-derived macrophages. These results are discussed in relation to the study of immune function in a novel species, and with particular reference to research into tuberculosis (Tb), since ferrets are important wildlife vectors of bovine Tb in New Zealand.     

In vitro effects of osteotomy angle and osteotomy reduction on tibial angulation and rotation during the tibial plateau-leveling osteotomy procedure

OBJECTIVE: To determine the effect of osteotomy angle, reduction technique, and tibial plateau rotation angle on angular and rotational limb deformities. STUDY DESIGN: Geometric comparison using bone models. METHODS: Rotational osteotomies were made in the proximal metaphysis of artificial tibias at 0 degrees, 10 degrees, 20 degrees, -10 degrees, and -20 degrees from perpendicular with respect to either the proximodistal and craniocaudal tibial axes. Negative-numbered angles represented osteotomies made from distal to proximal or caudal to cranial. Changes in tibial angulation and torsion were measured using a 3-dimensional digitizing instrument at tibial plateau rotation angles from 0 degrees to 30 degrees at 5 degrees increments. Two osteotomy reduction techniques were used: complete osteotomy reduction and alignment of the medial cortex. The mean of 5 measurements of torsional and angular tibial deformity for each of the 9 osteotomy orientations in each reduction technique group was obtained. RESULTS: All had increasing angular and rotational deformity as tibial plateau rotation angle increased. In the medially aligned cortex group, all tibias had valgus deformity, and 8 of 9 tibias were internally rotated. In the reduced osteotomy group, minimal angular deformity was seen in tibias with osteotomy variation along the proximodistal axis; however, tibias with osteotomy variation along the craniocaudal axis had angular deformity ranging from 6.0 degrees of varus deformity to 14.3 degrees of valgus deformity. Rotational deformity was affected similarly by osteotomy variation along either axis. Reduction technique had greater affect on angular and rotational deformity than osteotomy angle variation. CLINICAL RELEVANCE: These results suggest that osteotomy reduction may play a greater role in angular and rotational deformity than osteotomy angle, although extreme osteotomy angles should be avoided. To decrease the severity of deformity, we recommend that the osteotomy be made perpendicular to the craniocaudal and proximodistal axes and be completely reduced with less regard for alignment of the medial cortex.