Supplementation of Plant-Based Cryopreservation Medium with Folic Acid Conserves the Quality of Bulk Post-Thawed Spermatozoa.

The current study evaluated the effects of cryopreservation medium supplementation with folic acid as an antioxidant on post-thawed semen quality in bulk. Semen samples were collected from four proved Iranian Mahabadi bulls and diluted in extender containing 1.5% soybean lecithin. The diluted semen was assigned into six parts and supplemented with different doses of folic acid as follows: FA0 (extender without folic acid), FA0.05, FA0.1, FA0.2, FA0.4, and FA0.8 (extenders containing 0.05, 0.1, 0.2, 0.4, and 0.8 mM folic acid, respectively). Then, the semen samples were cryopreserved in liquid nitrogen. Sperm motility and velocity parameters, membrane integrity, abnormal morphology, viability, and lipid peroxidation were evaluated after thawing. In the results, FA0.05 presented higher (p≤0.05) total motility, progressive motility, membrane integrity, and viability and lower lipid peroxidation compared to other groups. Abnormal morphology was not affected (p>0.05) by treatments. In conclusion, supplementation of cryopreservation medium with 0.05 mM folic acid is a helpful method to conserve the quality of post-thawed semen in bulk.

The effect of dietary coenzyme Q10 on plasma metabolites and hepatic gene expression in broiler breeder hens.

1. This study was performed to evaluate the effects of dietary supplementation of coenzyme Q10 (CoQ10) on laying rate, body weight, plasma metabolites and some liver gene expression in broiler breeder hens. 2. A total of 128 broiler breeder hens (Arbor Acres Plus, 47 weeks of age) were randomly distributed to four dietary groups supplemented with different levels of CoQ10 (0, 300, 600 or 900 mg/kg diet) with four replicates of eight hens each. During 47-54 weeks of age, laying rate, egg mass and body weight were recorded weekly. To assay plasma biochemical indicators, blood samples were collected at 54 weeks of age. At the end of the experiment, for evaluating the abdominal fat weight, liver weight and expression of the adiponectin and proliferator-activated receptor-α (PPAR-α) genes in the liver, eight hens per treatment were selected, weighed and humanely killed by decapitation. 3. Dietary supplementation of CoQ10 linearly decreased abdominal fat weight, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities by increased levels of CoQ10. The plasma levels of glucose, cholesterol and alkaline phosphatase (ALP) activity were quadratically decreased by increased levels of CoQ10. The best plasma levels of glucose, cholesterol and ALP activity were estimated at 562.5, 633.3 and 517.8 mg CoQ10/kg diet, respectively. Adiponectin and PPARα gene expression exhibited a linear increased by increased levels of CoQ10. 4. In conclusion, addition of CoQ10 to the diet influenced lipid metabolism and expression of the adiponectin and PPAR-α genes, which might be partially due to the improvement in mitochondrial metabolism and energy production. However, further studies are necessary to determine the effects of CoQ10 on these indicators in broiler breeder hens during ageing.

Lecithin nanoparticles enhance the cryosurvival of caprine sperm.

This study was designed to compare the effects on goat spermatozoa cryosurvival of nano-lecithin-based (NL), lecithin-based (L) and egg yolk-based (EY) extenders. Ejaculates were collected from four fertile goats using artificial vagina and diluted with nine extenders. NL and L were tested at concentrations 1%, 2%, 3% and 4% (w/v), versus 15% (v/v) egg yolk-based extender. Overall, sperm quality (higher motility, viability and HOST, and lower apoptosis) was higher for NL than for L treatments (P < 0.05 for most cases, except for 1%). NL at 1% and especially at 4% showed lower motility and viability than 2% or 3% NL. NL at 2% achieved a better performance (P < 0.05) than EY for VCL (131.5 ±â€¯1.3 vs. 120.3 ±â€¯1.9 µm/s), VSL (43.9 ±â€¯1.5 vs. 35.8 ±â€¯1.4 µm/s), LIN (35.7 ±â€¯0.6 vs. 29.3 ±â€¯0.8%), WOB (47.0 ±â€¯0.5 vs. 43.9 ±â€¯0.9%) and viability (66.4 ±â€¯1.7 vs. 52.7 ±â€¯1.9%). Late apoptotic spermatozoa were also lower in 2% NL compared to EY (16.0 ±â€¯0.5 vs. 26.3 ±â€¯1.1%, P < 0.001). EY and 2% NL were compared in an IVF trial, with no significant differences in cleavage (68.8 vs. 70.8%) or blastocyst ratios (21.3 vs. 20.8%). In conclusion, using 2% nanolecithin in semen dilution could improve sperm cryosurvival of goat.

Effects of dietary curcumin supplementation on seminal quality indices and fertility rate in broiler breeder roosters.

1. Decreased semen quality is an underlying contributor to age-related subfertility in broiler breeder roosters. This study investigated the effects of dietary curcumin (derived from turmeric) supplementation as an antioxidant source on semen quality and fertility in broiler breeder roosters. 2. Twenty-eight Ross 308 roosters were randomly allotted to four groups with seven birds in each and were fed a standard diet supplemented with different levels of curcumin at 0 (C0), 10 (C10), 20 (C20) and 30 (C30) mg/bird per day from 48 through to 61 weeks of age. Body weight and semen quality traits were evaluated on a weekly basis and seminal concentrations of malondialdehyde (MDA) as a measure of antioxidation status were quantified at one-week intervals during the first 11 weeks of the trial (48-59 weeks of age). Semen samples from last 2 weeks (60 and 61 weeks of age) were used to artificially inseminate to assess the sperm-egg penetration (SP) in perivitelline membrane and fertility rates. 3. Except for body weight and ejaculate volume, other characteristics, including semen concentration, total sperm production, progressive motility and plasma membrane integrity were linearly improved by the increasing levels of curcumin supplementation (P < 0.01). However, dietary curcumin levels were linearly and quadratically associated with decreased seminal concentration of MDA (P < 0.01 and P < 0.03), percentage of abnormal sperm (P < 0.01 and P < 0.07) and increased plasma membrane functionality (P < 0.01 and P < 0.04), respectively. The SP holes in perivitelline membrane were increased in a linear and quadratic manner in response to increasing levels of curcumin (P < 0.01). Moreover, fertility rate was linearly improved (P < 0.01) as the dosage of curcumin increased, and resulted in 8, 12 and 14% improvements in the birds fed C10, C20 and C30, compared to C0, respectively. 4. In conclusion, the results showed that increasing levels of dietary supplementation of curcumin was associated with beneficial effects on semen quality indices and fertility rate in aged broiler breeder roosters.

Effects of supplemental conjugated linoleic acids (CLA) on fresh and post-thaw sperm quality of Holstein bulls.

This study was designed to investigate the effects of feeding-protected conjugated linoleic acid (CLA) on the semen production and sperm freezability in Holstein bulls. Twelve bulls were randomly assigned to two groups (n = 6 per group). Bulls received the normal diet (control group) or the normal diet top-dressed with 50 g of CLA (treated group) for 10 weeks. The control group received 40 g/day calcium soap of fatty acid. Fresh and post-thaw semen quality was assessed on ejaculates collected at the 0, 4, 6, 8 and 10 week of supplementation. Semen evaluations including sperm concentration, motion characteristics (subjective and computer-assisted), viability (Eosin-Nigrosin), membrane integrity (hypo-osmotic swelling test) and abnormality were conducted. Semen volume, sperm concentration and total sperm output were not affected by dietary treatment (p > .05). The proportion of spermatozoa with abnormal morphology in fresh semen significantly increased (p < .05) in the CLA-fed group compared to control group. Also, in CLA-fed group, the proportion of post-thaw spermatozoa with abnormal morphology at week 10 of trial was significantly higher in CLA than control group (p < .05). Progressive motility tended to be increased in the CLA-fed group, although dietary supplementation did not affect other CASA parameters or viability in fresh and frozen-thawed sperm. In this study, CLA supplementation had little positive effect on fresh or post-thaw sperm quality of Holstein bulls.

Plasma concentrations of PGFM and uterine and ovarian responses in early lactation dairy cows fed omega-3 and omega-6 fatty acids.

A total of 120 dairy cows were assigned randomly to three diets to determine the effects of omega-6 or omega-3 fatty acid (FA) supplementation on uterine diseases, ovarian responses, and blood concentrations of estradiol, progesterone, and PGFM in lactating Holstein dairy cows. Diets contained either protected palm oil (C), extruded linseed (L), or roasted whole soybeans (S), and they were fed from calving to Day 70 postpartum. Estrous cycles were synchronized and ovarian follicular development was monitored daily for an entire cycle. There were no differences among diets in the incidence of lameness, mastitis, or metritis, but the incidence of clinical endometritis was lower (P < 0.05) in cows fed S (0%) compared with cows fed C (28.2%) and L (20.5%). Uterine involution in cows fed S occurred 3.77 and 2.78 days earlier, respectively, than in those fed C and L. The PGFM response 60 minutes after an oxytocin challenge was highest for cows fed S and lowest for cows fed L. Mean plasma progesterone concentration on Day 15 of the synchronized cycle was higher in cows fed S (14.5 ng/mL) and L (15.0 ng/mL) than in those fed C (12.0 ng/mL). The ovulatory follicle on Day 21 of the estrous cycle (estrous = Day 0) was larger in cows fed S (16.1 ± 0.9 mm) and L (15.7 ± 0.7 mm) compared with cows fed C (13.2 ± 0.87 mm; P = 0.02) but there were no significant differences between cows fed diets S and L. The mean number of small and medium follicles and diameter of subordinate follicle were similar among diets. In conclusion, feeding a source of omega-6 FA can be a strategy to improve uterine health after calving, although a source of omega-3 FA such as L should be fed after uterine involution to decrease PGF2α secretion.

Effects of different polyunsaturated fatty acid supplementations during the postpartum periods of early lactating dairy cows on milk yield, metabolic responses, and reproductive performances.

In spite of the difficulties in delivering PUFA to ruminants, studies have generally indicated that the PUFA of the omega-6 (linoleic acid) and omega-3 [α-linolenic acid; eicosapentaenoic (EPA), C20:5 omega-3; docosahexaenoic (DHA), C22:6 omega-3] families are the most beneficial to improving reproduction in cows. The objectives were to determine if a diet enriched in α-linolenic acid (omega-3) or linoleic acid (omega-6) would influence milk production and composition, metabolic status, and reproductive performance in lactating dairy cows. High-yielding multiparous Holstein dairy cows (n = 120) with no overt clinical illnesses were blocked according to calving date and parity. Cows were assigned randomly to be fed 1) soybean whole roast (Soy, omega-6, n = 40) or 2) linseed (Lin, omega-3, n = 40) or 3) palm oil as a source of SFA (PO, n = 40) from calving until first heat after 40 d postpartum (dpp), and then half of the cows in each treatment group were switched to receive either Lin or SFA (PO) from first heat after d 40 to 120 dpp. Blood was collected from a subsample of cows. Blood was collected at 14 d intervals for 12 wk, starting on the day of calving. Results showed milk yield and DMI were not affected. Milk compositions were similar (P > 0.08) among diets, except concentration and yield of milk fat percentage, which was less in cows fed Lin (P < 0.05). Uterine involution in cows fed Soy occurred earlier (P < 0.05). Diets affected day to first estrus and day to first insemination in cows (P < 0.05). There were no differences among treatments for percent heat detection, percent pregnancy per first insemination, and percent conception per AI at estrus. Also, there is a trend of pregnancy by 120 d, which is 66.7% for the Lin group vs. 50.91% for the PO group (P < 0.08). Of the 4 pregnancy losses, 2 occurred in PO-PO group and 2 occurred in Soy-PO group, and none occurred in the other 4 treatments. In conclusion, our study showed feeding omega-6 fatty acids during 40 dpp could be a good treatment for early postpartum periods, and a shift to omega-3 fatty acids until 40 d after AI can be considered as a strategy for improving fertility in lactating dairy cows.

Performance, carcass traits, muscle fatty acid composition and meat sensory properties of male Mahabadi goat kids fed palm oil, soybean oil or fish oil.

This study examined the effect of palm, soybean or fish oils on the performance, muscle fatty acid composition and meat quality of goat kids. Twenty-four male Mahabadi kids (BW=19.4±1.2 kg) were divided into three groups according to liveweight and randomly allocated to one of three diets. Animals were fed ad libitum for 84 days. Different dietary fat sources had no effect on performance and/or carcass quality attributes. The soybean oil diet decreased 16:0 and 18:0 concentrations and increased 18:2 and 18:3 and the ratio of PUFA/SFA in the muscle compared with other treatments. Fish oil feeding increased 20:5 n-3 and 22:6 n-3 concentrations and decreased the ratio of n-6/n-3 in the muscle. The results demonstrate that the use of fish oil is a nutritional strategy to improve the health claimable long-chain omega-3 fatty acid content and n-6/n-3 ratio in goat meat without changing the sensory properties or colour of meat.

Combined effect of DHA and α-tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition.

The aim of this study was to investigate the effect of adding n-3 fatty acids (FA) and α-tocopherol (VE) to semen extender on freezing ability and FA composition of bull sperm. Semen was collected from 10 Iranian Holstein bulls and was pooled. In the first experiment, semen was divided into 12 groups including 4 levels of n-3 FA (0, 0.1, 1, 10 ng ml(-1) ) and 3 levels of VE (0, 0.1, 0.2 mmol). The treatment of 0.2 mmol VE and 10 ng ml(-1) n-3 FA had the best post-thawed sperm characteristics (P < 0.01). In the second experiment, lipid composition of the latest treatment and control (without FA and VE) was determined. Adding n-3 FA increased docosahexaenoic acid (DHA) percentage before freezing and after thawing. The ratio of n-3 to n-6 before freezing was higher (P < 0.05) in treated group than in control, and this ratio in the fresh sperm was greater than in the post-thawed sperm (P = 0.1). Results suggested that adding DHA accompanied with an antioxidant to an extender could improve cryosurvival of bull sperm via altering membrane lipid composition.

A signaling role for leptin in puberty onset in female rats?

The brain might initiate puberty in response to adequate leptin signaling from the periphery. We studied the link between whole body fat, plasma leptin levels, and puberty onset, in both controls and food-restricted female Wistar rats from age 22 to 42 days. Body fat correlated positively with the prevailing plasma leptin levels (r = 0.776) and with the time of puberty onset, i.e. vaginal opening (VO) (r = 0.691). Blood samples collected every other day at ZT 2, 6, and 12, showed a diurnal rhythm in leptin levels with a nadir at ZT 6. Furthermore, leptin levels increased over the pubertal period. Food restriction (FR) delayed the time of VO considerably (median VO at 38 vs 28 d), and body fat and plasma leptin levels were lower in the FR group (p <0.01), although the positive correlation between body fat and leptin levels remained. Only the absolute, but not the relative amount of body fat increased with age. These data support the notion that leptin could indeed serve as the link between nutritional status and the reproductive axis, and in this way participate in the timing of puberty.

Central application of IGF-1 postpones time of vaginal opening in normally fed, but not in food-restricted rats.

BACKGROUND/AIMS: Central but also peripheral IGF-1 is suggested to play a role in the initiation of puberty as it directly affects GnRH synthesis and release. A possible intermediate in the effects of IGF-1 on puberty might be the adiposity-signaling hormone leptin, whose plasma levels are decreased in food-restricted (FR) rats. METHODS: IGF-1 was chronically centrally infused in 23-day-old prepubertal female rats which were either normally fed or 30% FR, and the effects on time of vaginal opening (VO) and plasma leptin levels were monitored. RESULTS: FR treatment postponed time of VO and decreased plasma leptin levels. In normally fed rats centrally infused with IGF-1, time of VO was found to be postponed to the same extent as FR treatment did. The IGF-1 infusion did not affect plasma leptin levels in normally fed animals but increased leptin levels in the FR group compared to controls. Daily food intake was equal between all groups but body weight course was lower in FR rats. IGF-1 treatment did not significantly affect food intake or body weight course. CONCLUSION: FR treatment delays the moment of vaginal opening to the same extent as observed in normally fed rats that were centrally infused with IGF-1.

Chronic leptin infusion advances, and immunoneutralization of leptin postpones puberty onset in normally fed and feed restricted female rats.

Does leptin play a vital role in initiating puberty in female rats and can it overrule a nutrionally imposed (i.e. a 30% feed restriction, FR) delay in puberty onset? Prepubertal female rats were chronically infused for 14 days with leptin (icv or sc) or leptin-antiserum (icv) while puberty onset was monitored by means of scoring the moment of vaginal opening (VO). Median VO age was higher (35 days versus 27 days) in FR animals but leptin levels at VO were significantly decreased (1.44 +/- 0.17 ng/ml versus 2.79 +/- 0.31 ng/ml). Centrally (icv) and peripherally (sc) infused leptin (1 microg/day) advanced VO age compared to FR controls (30 days versus 35 days and 31 days versus 41 days, respectively). Congruently, centrally (icv) administered leptin-antiserum (0.6 microg/day) delayed puberty onset. In normally fed rats median VO age was only marginally advanced (26 days versus 27 days) but only if leptin was applied centrally. The effects of FR on puberty onset are counteracted or even normalized by the infusion of leptin, whereas immunoneutralization of central leptin postpones puberty onset. We therefore conclude that central leptin is crucial for initiating puberty in female rats.