A cooperative study assessing reproductive performance in sows fed diets supplemented with organic or inorganic sources of trace minerals.

Sows from three university research facilities (n = 245) were stratified by parity and initial body weight (BW), and within outcome groups, randomly assigned to fortified corn- and soybean meal-based control or organic trace mineral-supplemented, gestation (3,339 kcal/kg ME; 0.62% standradized ileal digestible [SID] lysine), and lactation (3,374 kcal/kg ME; 0.97% SID lysine) diets. Control gestation and lactation diets were supplemented with inorganic trace minerals (120 ppm Zn from ZnO, 30 ppm Cu from CuSO4, and 50 ppm Mn from MnSO4), and the experimental diets contained the same total level of minerals but complexed organic trace minerals replaced 50% of the inorganic trace minerals. Sows were fed to condition during gestation and on an ad libitum basis during lactation. Sow BW (breeding, d 110 of gestation, 48 h post-farrowing, and weaning) and feed consumed were recorded. During gestation, control sows tended to gain less weight (60.4 vs. 64.6 kg, P = 0.06) and consumed less feed (263.5 vs. 264.8 kg, P = 0.05), and had poorer Gain:Feed (G:F) (0.27 vs. 0.29, P = 0.04) than sows fed the organic trace minerals. Sow average daily feed intake (ADFI) during lactation was similar (P = 0.28) between groups (4.93 vs. 4.74 kg for control and treated sows, respectively). Number of pigs born alive (11.4 vs. 10.9, P = 0.24) and weaned (10.2 vs. 9.8, P = 0.18), and pig pre-weaning average daily gain (ADG) (0.27 vs. 0.27 kg/d, P = 0.77) and mortality (13.1 vs. 12.9%, P = 0.92) were similar for control and treated sows, respectively. Results of the current study demonstrate that sows fed diets supplemented with organic trace minerals displayed similar reproductive performance, but improved weight gain and G:F during gestation compared with sows fed inorganic trace minerals.

Handler training improves decontamination of working canines with oil-based exposure in field conditions using disposable kits.

Exposure to contaminants in the field is a reality for deployed canines. To date, there is no data evaluating the benefits of training for handlers associated with canine decontamination efforts. The objective of our work was to investigate the impact of handler training on the reduction of oil-based contaminants in working canines. Canine teams (n = 10) were randomly assigned to either TRAINED or UNTRAINED groups. Each team (handler and dog) in the TRAINED group received 30-minutes of interactive training using an illustrated guide on proper utilization of equipment provided. Teams in the UNTRAINED group received the same equipment and illustrated guide but no interactive training. Decontamination efforts were measured using an oil-based pseudo-contaminant (GloGerm®, Moab, UT) topically applied to four anatomical locations: cranial neck, between the shoulder blades, left medial hindlimb and left hind paw with pre- and post-washing images collected from a fixed distance of 20 inches. Visual assessment of contaminant reduction was scored as follows: 0 = <24% contaminant reduction; 1 = 25-50% contaminant reduction; 2 = 51-75% contaminant reduction; and 3 = >76% contaminant reduction. No score discrepancies >1 were reported between reviewers. Trained handlers were more effective at contamination reduction (P = .0093) as compared to their untrained counterparts. These results indicate that handlers, when properly trained, can achieve reduction of oil-based contaminants with a disposable decontamination kit and a garden hose.

Effect of Catalase or Alpha Lipoic Acid Supplementation in the Vitrification Solution of Ovine Ovarian Tissue.

AIM: The present study evaluates the effect of different concentrations of antioxidants (catalase - CAT and alpha lipoic acid - ALA) on the follicular activation and morphology, DNA damage, ROS production, and mitochondrial activity in vitrified sheep ovarian tissue. METHODS: This experiment was divided into two steps. First, ovarian fragments were distributed into the following treatments: fresh tissue or control (CTR), incubation (INC), vitrification without antioxidant (VWA), with CAT (10, 20, or 40 IU mL-1) or ALA (25, 50, or 100 µM mL-1). After vitrification/warming, the fragments were additionally incubated for 24 hours and evaluated for morphology and follicular activation, as well as reactive oxygen species (ROS) levels in the culture medium. For the second step, other ovarian fragments were submitted to CTR, VWA, CAT40, and ALA100. After vitrification/warming, the fragments were incubated for 24 hours and evaluated by cell density of ovarian stroma, DNA damage, and mitochondrial and intracellular ROS levels. RESULTS: The percentage of morphologically normal follicles in vitrified ovarian tissue in the presence of ALA in all concentrations did not differ (p > 0.05) from fresh tissue or CTRs. The percentage of activated follicles was higher in ALA100 µM mL-1 than those observed for the treatments INC, CAT (40 IU mL-1), or ALA (25 or 50 µM mL-1). The use of CAT affected (p < 0.05) the density of stromal cells (40 IU mL-1), ROS levels (10 and 20 IU mL-1), as well as DNA damage revealed by ©H2AX (40 IU mL-1). CONCLUSIONS: Although 100 µM/mL of ALA did not alter intracellular ROS, this concentration reduced the levels of ROS in the culture medium, preserved both the follicular morphology, as well as the mitochondrial activity, promoted follicle activation, and protected the follicles from DNA damage.

Sheep Isolated Secondary Follicles Are Able to Produce Metaphase II Oocytes After Vitrification and Long-Term In Vitro Growth.

The vitrification of preantral follicles followed by in vitro growth (IVG) could be valuable to produce fertilizable oocytes. However, the meiotic resumption rates of oocytes cultured from vitrified secondary follicles (SF) have been reported as suboptimal. This study aimed to verify two base media (alpha modification of minimum essential medium, α-MEM, and tissue culture medium 199, TCM199) on vitrified SF regarding different requirements during IVG. Sheep ovarian fragments were divided in six groups: (1) Fresh groups (Control α-MEM and TCM199): SF without vitrification; (2) Follicle-Vitrified (Follicle-Vit α-MEM and TCM199): SF vitrified after isolation; and (3) Tissue-Vitrified (Tissue-Vit α-MEM and TCM199): SF vitrified enclosed in ovarian fragments and, subsequently, isolated. The isolated SF were submitted to IVG for 18 days. Thereafter, the recovered cumulus-oocyte complexes (COCs) underwent in vitro maturation (IVM) and evaluation of chromatin configuration. Follicular granulosa cells were analyzed for their gene expression of Bax, Bcl2, and Connexins (CX) 37 and 43. COCs from in vivo antral follicles were used as in vivo control. Data were analyzed by analysis of variance, Tukey, and chi-square tests. Differences were considered significant if p-value is <0.05. Follicle-Vit groups had higher (p < 0.05) percentage of antrum formation compared with Tissue-Vit groups. Vitrification did not affect (p > 0.05) oocyte diameter postmaturation. Oocytes from Follicle-Vit in α-MEM reached metaphase II stage after IVM. Gene expression for CX37, CX43, and Bax was lower in Tissue-Vit groups. For Bcl2, the gene expression was the opposite. In conclusion, during IVG for 18 days, maximal oocyte meiotic resumption was not negatively impacted by vitrification and was greatest for isolated SF using α-MEM as a medium.

Accelerated follicle growth during the culture of isolated caprine preantral follicles is detrimental to follicular survival and oocyte meiotic resumption.

This study investigated the effect of androstenedione (A4) alone or in association with different concentrations of bovine recombinant FSH on the IVC of isolated goat preantral follicles. Follicles were mechanically isolated from ovarian tissue and cultured for 18 days in α-minimum essential medium supplemented or not with A4 (10 ng/mL) alone or in association with fixed (A4 + FixFSH: 100 ng/mL) or sequential (A4 + SeqFSH: Day 0, 100 ng/mL; Day 6, 500 ng/mL; Day 12, 1000 ng/mL) concentrations of FSH. After 18 days, the oocytes were recovered for IVM and fluorescence analysis. At Day 18 of culture, only A4 + SeqFSH treatment showed a lower (P < 0.05) rate of intact follicles, survival probability, and meiotic resumption, as well as higher (P < 0.05) percentage of degeneration and/or extrusion after antrum formation. Taken together, these results reported a positive correlation between fast-growing follicles and follicles that degenerated and/or extruded after antrum formation. When compared with control, the addition of A4 alone or in association of FSH did not increase (P > 0.05) the estradiol production or androstenedione levels on Day 6. However, on Day 18, the androstenedione levels were significantly lower in A4 + SeqFSH treatment when compared with A4 alone or to A4 + FixFSH treatments, whereas the estradiol production did not differ (P > 0.05). In summary, this study found that accelerated follicle growth negatively impacted the morphology of caprine preantral follicle cultured in vitro. In addition, the association of androstenedione with increasing concentration of FSH was detrimental to follicular survival and oocyte meiotic resumption.

Equine spermatozoal motility and fertility associated with the incorporation of d-(+)-mannose into semen extender.

Mannose is capable of decreasing bacterial attachment to the uterine mucosa in mares. Bacteria gain entry into the mare's uterus during breeding; therefore, a practical method to deliver mannose to the uterus is to incorporate it into semen extenders. The effect of mannose on spermatozoal motility and subsequent sperm fertilizing capability is unknown. The present study evaluated progressive spermatozoal motility in semen extender formulations incorporating mannose and assessed the fertility of mares inseminated with a mannose-containing semen extender. In Experiment 1, progressive spermatozoal motility in extender mixtures containing 0 mannose (control), 25, 37 or 49 mg/mL mannose was evaluated at 20 degrees C or 5 degrees C holding temperatures for 0, 12, 24 and 48 h post-dilution. Measures were repeated three times using five stallions of proven fertility. High concentrations of mannose in the extender affected progressive motility beyond the time and temperature effects noted in the controls. Extender containing only mannose sugar (49 mg/mL) displayed an immediate depression in progressive motility compared with controls (45.5% versus 62.9%, respectively; P<0.001). The 37 mg/mL mannose extender had a less dramatic decrease in motility (P<0.05) and only after storage at 5 degrees C for > or =12h (48.7% versus 58.0%, respectively). Extender with 25 mg/mL mannose performed no differently than the control formulation under all conditions. In Experiment 2, two groups of mares (n=11 each) were inseminated with 500 x 10(6) progressively motile spermatozoa extended in a traditional skim milk (control) extender or the 37 mg/mL mannose extender preparation. A single-cycle pregnancy rate of 72% was achieved by both groups. Present data suggest that a semen extender containing up to 37 mg/mL mannose could maintain motile spermatozoa for on-farm use and 25 mg/mL mannose concentrations preserved motility during long-term cooling. Likewise, sperm extended with up to 37 mg/mL of mannose had the same fertilizing capability as sperm in traditional extender mixtures.