BACKGROUND: Nowadays, the interest in functional food has dramatically increased. Herbal plants and functional foods have health-enhancing effects on consumers due to their medicinal, antioxidant, and nutritional properties. Probiotics are one of the most emerging and popular functional food products. OBJECTIVES: This study aims to assess the effect of irradiated and non-irradiated saffron petal extract on the viability of Lacticaseibacillus paracasei (M4PM99) in probiotic yogurt.METHODS The ethanolic extract of irradiated saffron petals with a 10 KGy dose of gamma ray at concentrations of 25, 50, and 75 mg/mL and non-irradiated extract at the same concentrations were used and their effect on the viability of Lacticaseibacilus paracasei and their antioxidant and physicochemical properties in set yogurt were studied. Probiotic survival, pH, acidity, content of total phenolic compounds, DPPH inhibition percentage, and sensory properties on days 0, 7, 14, and 21 were assessed.RESULTS: Both irradiated and non-irradiated saffron extracts significantly increased the viability of probiotic bacteria compared to the control sample (P<0.05). The addition of extracts was effective in increasing acidity and decreasing pH compared to the control (P<0.05). With the increase in the amount of extract, the percentage of DPPH inhibition and phenolic compounds significantly increased in the irradiated samples (P<0.05). The effect of storage time was also significant on these indicators, such that the antioxidant properties and phenolic compounds increased until the 14th day and then decreased (P<0.05). In the sensory evaluation, in terms of taste, odor, and color, the lowest score was related to the sample containing 0.75% extract. No significant difference was observed in other concentrations compared to the control sample.CONCLUSIONS: Saffron petal extract has a positive effect on the viability of probiotics during storage. Gamma irradiation has a significant effect on the amount of total phenolic compounds and the antioxidant activity of saffron petal extract. It can be used as a natural antioxidant in dairy products.

BACKGROUND: Storing semen at a temperature of 4 degrees Celsius reduces its motility and quality.OBJECTIVES: This study aimed to determine the effect of adding calcium compounds, including calcimaphor (CMP), calcium gluconate (CG), and calcium chloride (CaCl2), on the motility and progressive motility of rooster sperm kept at refrigerator temperature.METHODS: This research used five pieces of 45-week-old Lari breed roosters. The liquid diluent added different calcium compounds at 0.56, 0.056, and 0.0056 mM concentrations. After treatment, the diluted seminal samples were cooled at the storage temperature to avoid thermal doubt and then transferred to the refrigerator at 4 degrees Celsius. Parametric factors that were more important such as the percentage of laterality and progressive mobility, were measured visually using the lens of a 40-light microscope, survival was checked using the eosin-nigrosin staining method, acrosome health percentage by formalin citrate method, cell membrane health with hypoosmotic test, lipid peroxidation level, fertility was evaluated using perivitelline membrane sperm reaction 72 hours after storing at 4 degrees Celsius.RESULTS: Based on the results, different calcium compounds in most concentrations could significantly affect the parameters of survival, mobility, and progressive mobility (P<0.05).CONCLUSIONS: Most of the sperm-related parameters in the control group decreased after this period of storage in the refrigerator, but the addition of calcium gluconate (0.56, 0.056, and 0.0056), Calcimaphor (0.56, 0.056, and 0.0056) and calcium chloride (0.56, 0.056 and 0.0056) to the semen thinner maintained the quality indicators of rooster sperm.

Free, autogenous, full-thickness skin grafts were applied to 10 dogs; 5 dogs were given an iron chelator, deferoxamine-10% hydroxyethyl pentafraction starch (DEF-HES; 50 mg/kg of body weight, IV), and 5 dogs were given an equal volume of 10% hydroxyethyl pentafraction starch (HES) in 0.9% saline solution (5 ml/kg, IV). All dogs (DEF-HES/HBO- and HES/HBO-treated) were exposed to 60 minutes of hyperbaric oxygen (HBO) at 2 atmospheres absolute pressure twice daily for 10 days, beginning the day of surgery. The percentage of viable graft on day 10 was lower in HES/HBO-treated-dogs (mean +/- SD, 13.3 +/- 21.3%; median, 3.0%) than in DEF-HES/HBO-treated dogs (64.7 +/- 39.2%; 88.3%; P = 0.095, Mann-Whitney two-tailed test). There was a positive correlation between percentage of viable graft (on day 10) and percentage of haired skin on the graft site (on day 28) for all dogs (r = 0.91) and for HES/HBO-treated dogs (r = 0.97). The DEF-HES/HBO-treated dogs had less consistent correlation (r = 0.67). Perivascular aggregates of foamy cells were observed in the superficial and reticular portions of the dermis and in the subcutaneous tissue on both surfaces of the panniculus muscle in the graft sites of DEF-HES/HBO-treated dogs. These cells were also observed in the dermis, but not subcutaneous tissue of the control skin sections, and in some viscera of DEF-HES/HBO-treated dogs. Deferoxamine appears to attenuate the detrimental effect of HBO and HES on survival of free skin grafts. However, clinical use of HBO is not recommended as adjunct treatment for free skin grafts in dogs in the first 10 days after grafting. Administration of DEF-HES is not recommended because it has failed to improve the survival of free skin grafts, and the consequence of the cellular response seen in this study is undetermined.

Transcutaneous oxygen (PO2.TC) monitoring is commonly used in human medicine for evaluating skin viability. The application of transcutaneous monitoring for evaluating skin viability in dogs was investigated. The changes in PO2-TC values were measured from 16 avascular skin flaps created along the lateral hemithoraces of 4 dogs. Transcutaneous oxygen values were serially recorded from the vascular base and avascular apex of each flap for 12 hours after surgery. A single transcutaneous measurement was obtained from each flap base and apex 24 hours after surgery. Serial arterial blood gas analyses were obtained to compare central oxygen values with PO2-TC values. Full-thickness skin biopsy specimens were harvested from the base and apex of each flap 24 hours after surgery. The flaps were observed for 4 days and then excised for histologic examination. A subjective grading scale was used to assess histologic changes. Throughout the 12-hour period and at 24 hours, a statistically significant difference was found between the PO2-TC values for apices and bases of the flaps. The mean PO2-TC for all bases was 90.9 mm of Hg +/- 3.3 SEM, and the mean PO2-TC for all apices was 21.2 mm of Hg +/- 1.8 SEM. The mean regional perfusion index (apex PO2.TC/base PO2-TC) was 0.23 +/- 0.02. The subjective numbers assigned to the biopsy specimens were statistically evaluated by using a paired Student's t test and a Wilcoxon signed-rank test. A significant difference was found between the numbers for the collective bases and apices with both tests. A statistically significant difference was found between the numbers for the apex biopsy specimens taken 24 hours after creation of the skin flap and those taken when the flap was excised, whereas no difference was found between the numbers for the base biopsy specimens. On the basis of our findings, PO2-TC monitoring is a useful technique for assessing skin viability in dogs.

Effects of the endophyte Acremonium coenophialum in tall fescue on pregnant mares and foal viability were evaluated. Twenty-two mature pregnant mares were randomly chosen to graze either Kentucky-31 tall fescue that was free from A coenophialum (endophyte-free, EF) or tall fescue infected with A coenophialum (endophyte-present, EP) after the first 90 days of pregnancy through parturition. Concentrations of pyrrolizidine and ergopeptine alkaloids were significantly greater in EP grass, compared with EF pasture. Ten of 11 mares grazing EP pasture had obvious dystocia. Mean duration of gestation was significantly greater for the EP group, compared with the EF group. Foal survivability was severely reduced among mares grazing Ep fescue with only 1 foal surviving the natal period. Udder development and lactation were low in mares grazing EP grass. The absence of clinical problems in mares grazing EF grass implicated the endophyte as the causative agent of reproductive problems and perinatal foal mortality in pregnant mares grazing endophyte-infected fescue grass. Caution should be exercised in allowing pregnant mares to graze pastures infected with the endophyte A coenophialum.

Corynebacterium pseudotuberculosis phospholipase D (PLD) significantly affected viability of ovine neutrophils after 24 hours' exposure, This effect was more marked in cells that ingested PLD emulsified in oil. Treatment of neutrophils with PLD significantly (P < 0.05) reduced the ability of these cells to migrate toward activated sheep serum. The PLD was not chemotactic, but it activated normal sheep serum, producing factors that were chemotactic for neutrophils.

Perfusion and viability of island axial pattern skinflaps were tested in 37 healthy New Zealand white rabbits, using laser Doppler monitoring of blood flow in the capillary loops and the subpapillary plexus of the dermis. Skin flaps, selected on the basis of the caudal superficial epigastric vein and artery, were lifted and replaced in their original locus after selective occlusion of their vascular pedicles. Subjects were allotted into groups: control group (n = 10); arterial occlusion (n = 7); venous occlusion (n = 10); and arterial and venous occlusion (n = 10). The rabbits were monitored from 48 hours before surgery until euthanasia 48 to 72 hours after replacement of the flap. Flap viability was assessed on a clinical basis, using a comparative scoring method based on a numeric scale. The degree of necrosis in histologic sections was evaluated, using a scoring system. Laser Doppler measurements were obtained on 3 consecutive days before surgery, to establish the normal basal blood flow in the skin. Postsurgical measurements were obtained at 2-hour intervals for the first 8 hours and at 24, 48, and 72 hours after surgery. Measurements of basal blood flow varied significantly (P < 0.05) from site to site on the surface of individual flaps and over time. When laser Doppler flowmetric (LDF) measurements from 6 sites on a flap were used as a measure of laser Doppler flow for the total flap, there was no significant difference between contralateral flap areas outlined on the abdomen of the rabbits. Temporal variations over 3 days for each rabbit or among rabbits were not significant. The LDF measurements detected acute vascular occlusion when compared with the controls, and were able to differentiate between control and arterial occlusion groups, control and venous occlusion groups, control and arterial and venous occlusion groups, arterial and venous occlusion groups, venous and arterial and venous occlusion groups (P < 0.05), but not between arterial and arterial and venous occlusion groups. Evaluation of LDF values at 4 hours proved to be a better predictor than clinical assessment at 4 or 8 hours in evaluating skin flap viability.

Demi-embryos were successfully produced by bisection of ICR mouse embryos at preimplantation stages. They were microsurgically bisected using a microsurgical blade attached to a micromanipulator after pretreatment with 0.5 % pronase in PBS for two minutes or not. Embryos with softened zona pellucida were more easily bisected and less damaged than intact embryos. The highest success rate in bisection has been achieved by selecting blastocysts (94.1 % in success rate with intact blastocysts and 100 % in success rate with zona softened blastocysts). Demi-embryos without zona pellucida were cultured in D-PBS or M-16 medium at 37deg C, 5 % CO2 in air for 72 hours for 2-cell stage embryos, 48 hours for 4-to 8-cell stage embryos, 24 hours for morula stage embryos and 6-12 hours for blastocyst stage embryos. For the in vitro culture of 2-cell stage embryos, 100 micro M 2Na-EDTA was added to the media. M-16 medium was better for the in vitro development of mouse embryos than PBS, and PBS is not considered to be suitable for long-term culture of embryos, especially at early stage of cleavage. In M-16 medium, developing rate of demi-embryos of which pair underwent development to form eublastocysts was 15.8 % at 2-cell stage, 16.8 % at 4-cell stage, 38 % at 8-cell stage, 89.6 % at morula stage and 94.4 % at blastocyst stage, respectively. The more rapid and efficient production of demi-embryos and higher viability after bisection can be expected by softening zona pellucida with pronase and by selecting morulae or blastocysts rather than embryos at early stage of cleavage.

Protocols for cooling or freezing goat semen usually recommend centrifugation for seminal plasma removal. However, little is known about the effect of this process on goat sperm viability and functionality. The present study evaluated the effects of centrifugation force on the plasma membrane, acrosomes, and DNA integrity of goat semen. Four ejaculates from each of the four different Anglo Nubian male goats were used. Semen samples were obtained using artificial vagina, and immediately after collection, ejaculates were diluted using Ringer’s sodium lactate solution and split into three groups: Control (CG, without centrifugation), G1 (centrifugation 600 x g/10 min), G2 (centrifugation 1200 x g/10 min). After centrifugation, seminal plasma was removed, the sperm pellets were resuspended using Tris-egg yolk extender (80 x 106 spermatozoa/mL) and the sperm morphology was analyzed. Samples were cooled at 5°C for 5, 24, 36, and 48 h and then sperm plasma membrane and acrosome integrity (PMAI, %) and sperm DNA fragmentation index (SDF, %) were evaluated at each time-point, using a flow cytometer. Additionally, sperm movement was determined using computer semen analysis (CASA) after 5, 24, and 48 h of refrigeration period. The semen centrifugation did not induce additional sperm morphology defect or reduction in sperm kinetics in the experimental groups. Differences were not observed (p > 0.05) in PMAI and SDF among different groups, in any of each timepointof the cooling process. In conclusion, centrifugation, even at high speeds, did not affect goat sperm integrity and functionality when submitted to refrigeration process.