What is the relevance of seminal plasma from a functional and preservation perspective?

When preserving sperm in the liquid or cryopreserved state, seminal plasma (SP) components within ejaculates can alter fertilizing capacity of these gametes. Depending on the species or how semen is collected, volume and concentration of SP components varies considerably. The SP contains substances essential for maintenance of sperm viability and fertility; however, these components can be deleterious depending on quantity, or duration of time before there is removal of SP from sperm in semen processing. Substances that impair (e.g., BSP - bull; HSP-1 - stallion; Major seminal plasma protein PSPI - boar) or improve (e.g., spermadhesin PSP-I - boar) spermatozoa fertilizing capacity have been identified. Depending on individual males, species, and semen collection procedures, SP removal may be beneficial before preservation in the liquid or cryopreserved state. In some cases, SP that is removed can be added back to thawing extender with there being positive effects in thawed sperm and for sperm viability in the female reproductive tract. In this review article, there is a focus on different effects of SP in samples of cooled and cryopreserved semen from four domestic species (pigs, horses, cattle, and sheep) with there being emphasis on how SP modulates the function and morphology of sperm cells before, during, and after preservation in the refrigerated or cryopreserved state. The present review is part of the Festschrift in honor of Dr. Duane Garner who made major contributions to the area of focus in this manuscript as evidenced by the many times his research is cited in this manuscript.

In vitro effects of two different commercial freezing and thawing extenders on boar sperm quality.

This study was conducted to evaluate whether there were differences in viability of cryopreserved semen when using two different freezing (Minitube Cryoguard - F1 or Androstar® CryoPlus - F2) and thawing (Minitube Cryoguard Thawing solution - T1 or Androstar® Plus - T2) extenders. Ejaculates were collected, diluted (1:1), and cooled before shipping at 17 °C overnight. Samples were aliquoted in cryopreservation extender F1 or F2. Four straws from each treatment sample were thawed and diluted in T1 or T2, resulting in four treatments (F1-T1, F1-T2, F2-T1, and F2-T2). The sperm in diluted semen were evaluated for motility kinetics at 30, 180, and 360 min after thawing. The integrity assessments of the plasma and acrosomal membranes were performed at 30 and 360 min after thawing. There was no interaction between F × T × Time (P > 0.05), and no interaction between F × T (P > 0.05). The sperm progressive motility (PMOT) as time post-thawing increased was greater (P = 0.015) when dilutions occurred using F1 compared with F2 extender. Sperm thawed in T1 had a greater TMOT (P = 0.008) and PMOT (P = 0.033) at all times evaluated. The sperm plasma and acrosomal membrane integrity (AIMI) were greater (P = 0.009) when samples were preserved in F1 compared to F2 extender. The use of T2, as compared with T1 thawing extender, resulted in an enhanced integrity of the plasma and acrosomal membranes (P = 0.008). It is concluded different combinations of commercial freezing extenders and thawing solutions have effects on the quality of cryopreserved boar semen in vitro.

A perspective on global access to insulin: a descriptive study of the market, trade flows and prices.

AIM: To describe the global insulin market. METHODS: Market intelligence data, United Nations Commodity Trade Statistics for insulin trade, the International Medical Products Price Guide for prices of human insulin and additional web searches were used as data sources. These sources were combined to gain further insight into possible links among market, trade flows and prices. Descriptive statistics and Spearman's rank order correlation were used for the analysis. RESULTS: A total of 34 insulin manufacturers were identified. Most countries and territories are reliant on a limited number of supplying countries. The overall median (interquartile range) government procurement price for a 10-ml, 100-IU/ml vial during the period 1996-2013 equivalent was US$4.3 (US$ 3.8-4.8), with median prices in Africa (US$ 4.7) and low- (US$ 6.9) and low- to middle- (US$ 4.7) income countries being higher over this period. The relationships between price and quantity of insulin (Spearman's r=0.046; P>0.1) and number of import links (Spearman's r=0.032; P>0.1) were weak. The links between price and percentage of total insulin from a country where a 'big three' manufacturer produces insulin (Spearman's r=0.294; P<0.05) and total insulin from the main import link (Spearman's r=-0.392; P<0.05) were stronger. CONCLUSIONS: This research shows the high variability of insulin prices and the reliance on a few sources, both companies and countries, for global supply. In addressing access to insulin, countries need to use existing price data to negotiate prices, and mechanisms need to be developed to foster competition and security of supply of insulin, given the limited number of truly global producers.

Differential gene expression of Eph-ephrin A1 and LEPR-LEP with high or low number of embryos in pigs during implantation.

The objective of this study was to ascertain whether mRNA and protein expressions of implantation-related genes (erythropoietin-producing hepatocellular receptor-ligand A1, Eph-ephrin A1 and leptin receptor-leptin, LEPR-LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre-pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO-NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion-allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high-embryo compared to the low-embryo group, respectively (p < .05). Real-time qPCR results showed that Eph-ephrin A1 mRNA expression was less in the high-embryo (p < .05) compared to the low-embryo group. In addition, Western blotting analysis indicated that Eph-ephrin A1 and LEP protein expression at endometrial attachment site in high-embryo was less (p < .05) compared to low-embryo group. It was also noted that mRNA expression of Eph-ephrin A1 and LEPR-LEP was greater in pregnant than non-pregnant gilts (p < .05). Moreover, mRNA expression of Eph-ephrin A1 (p < .05) and LEPR-LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph-ephrin A1, LEPR-LEP and embryo length with the correlation coefficient 0.31-0.59. For Eph-ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR-LEP, the highest correlation coefficient appeared between LEPR-LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph-ephrin A1 and LEPR-LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph-ephrin A1 and LEPR-LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph-ephrin A1 and LEPR-LEP might play roles in the regulation of embryo implantation in pigs.

Ovulation and fertility responses for sows receiving once daily boar exposure after weaning and OvuGel® followed by a single fixed time post cervical artificial insemination.

Boar exposure is used to stimulate follicle development and estrus in sows after weaning and also to improve semen uptake and sperm transport with insemination. However, the need and value of boar exposure is uncertain when ovulation induction is used. These studies were designed to determine the effect of daily boar exposure after weaning when used with ovulation induction and fixed time post-cervical artificial insemination (PCAI). In experiment 1, sows were weaned into stalls and assigned to receive 3 min of daily fenceline boar exposure (BE, n = 7) or no boar exposure (NBE, n = 8). All sows received OvuGel at 96 h after weaning and BE or NBE 30 min prior to a single PCAI 24 h after OvuGel. Ovaries were assessed daily for follicle size from weaning until ovulation. Cervical contractions were measured 30 min following BE or NBE and before PCAI, while uterine contractions were measured for 1 h following PCAI. In experiment 2, weaned sows (n = 244) were assigned by parity to receive once daily BE for 1.5 min each day or NBE. OvuGel, PCAI and ultrasound methods were performed similarly as in experiment 1. Results from experiment 1 indicated BE did not significantly influence follicle size or measures of fertility. However, BE did increase the frequency of cervical contractions (P < 0.05), but with no effect on the uterus. Results from experiment 2 indicated BE had no effect on catheter passage for PCAI but did increase the proportion of sows ovulating within 48 h after OvuGel (77.7 vs 67.5%, P = 0.05), and tended (P = 0.10) to increase the proportion of sows inseminated 24 h before ovulation (70.3 vs. 61.0%). However, BE had no effect on adjusted farrowing rate (84.4 vs. 77.4%) or total pigs born (13.2 vs. 12.5) for BE and NBE, respectively. There were treatment and parity interactions for follicle size at time of OvuGel and at time of PCAI (P < 0.05) with BE minimizing parity effects on follicle size. Parity effects were also evident on farrowing rate and litter size when inseminations occurred >24 h from ovulation but not when inseminations occurred ≤24 h before ovulation. The results indicate that boar exposure for only minutes each day after weaning had beneficial effects for improving follicle development, ovulation induction, and AI timing, most notably in parity 1 sows, but had no beneficial or detrimental effects on the ability to perform PCAI.

Design and biological effects of a vaginally administered gel containing the GnRH agonist, triptorelin, for synchronizing ovulation in swine.

A novel gel formulation was selected for intravaginal delivery of the GnRH agonist (triptorelin) for synchronizing ovulation in pigs. Studies with gilt models were used to assess LH response profiles. The lowest dose of triptorelin that induced the most gilts to show an LH surge was 100 µg in 1.2% methylcellulose gel. This formulation had a similar effect in weaned sows while also advancing ovulation. The timing of administration was evaluated in sows after weaning. Administration at 96 h induced more sows to ovulate (58%) by 48 h compared to treatment at estrus (45%) or for controls (34%), but the desired level of ovulation synchrony was not achieved. As a result, greater doses of triptorelin were tested and 200 µg given at 96 h after weaning, induced 81% of sows to ovulate within 48 h after treatment. The best synchrony of ovulation occurred when given at 96 h after weaning compared to earlier or later intervals. The optimum time to give a single fixed time AI (SFT-AI) after administration of 200 µg of triptorelin in 1.2% gel (OvuGel®) at 96 h after weaning was tested. A SFT-AI at 22 ± 2 h after OvuGel achieved the highest fertility and was practical for staff during the normal work day. In field trials, a SFT-AI 22 ± 2 h after all weaned sows were treated with OvuGel improved (P = 0.04) farrowing rate to 82.5% compared to control sows weaned (80.1%), with no effect on numbers of pigs born alive (12.1). Research continues for identifying the advantages for use of OvuGel in different production systems, and potential application for use in gilts.

Effects of estrus synchronization using Matrix® followed by treatment with the GnRH agonist triptorelin to control ovulation in mature gilts.

Estrus and ovulation responses in Matrix-treated gilts may affect ovulation synchrony in response to triptorelin. In experiment 1, estrus and ovulation measures at 12h intervals after last Matrix feeding (LMF) were analyzed. For the 398 gilts that displayed estrus, 87.4% were detected on Days 6-8 after LMF. Duration of estrus was 24-60h for 85.6% of gilts and negatively correlated with interval from LMF to estrus (r=-0.53, P<0.0001). The estrus to ovulation interval was positively correlated with duration of estrus (r=0.61, P<0.0001). In experiment 2, gilts (n=96) received intravaginal treatment with 2mL of gel containing placebo (Control) at 96h, 200µg of triptorelin at 96h (TRP96), 120h (TRP120) or 144h (TRP144) after LMF. Estrus measures did not differ (P>0.10) among treatments. The proportion of gilts ovulating 32-56h after treatment was greater for TRP120 and TRP144 (P<0.01) compared to other treatments. The treatment to ovulation intervals for all triptorelin treatments were shorter (P<0.001) than Control. In experiment 3, gilts (n=86) received placebo (Control), 100µg (TRP100), 200µg (TRP200), or 400µg (TRP400) of triptorelin at 120h after LMF. There was no effect of treatment (P>0.10) on estrus or on interval from LMF to estrus. The proportion of gilts ovulating by 40, 48 and 56h after treatment increased (P<0.05) with triptorelin compared to Control. Our results indicate that gilts receiving 100-400µg of triptorelin at 120h after LMF had the greatest ovulation synchrony 24-48h following treatment. These studies provide important information for developing a procedure for a single insemination in synchronized gilts.

Atypical neurofibromatosis type 1 with unilateral limb hypertrophy mimicking overgrowth syndrome.

Neurofibromatosis type 1 (NF1; OMIM 162200), a dominantly inherited multitumor syndrome, results from mutations in the Neurofibromin 1 (NF1) gene. We present the case of a Hungarian woman with the clinical phenotype of NF1 over her whole body and the clinical features of unilateral overgrowth involving her entire left leg. This unusual phenotype suggested either the atypical form of NF1 or the coexistence of NF1 and overgrowth syndrome. Direct sequencing of the genomic DNA isolated from peripheral blood revealed a novel frameshift mutation (c.5727insT, p.V1909fsX1912) in the NF1 gene. Next-generation sequencing of 50 oncogenes and tumour suppressor genes, performed on the genomic DNAs isolated from tissue samples and peripheral blood, detected only wild-type sequences. Based on these results, we concluded that the patient is affected by an unusual phenotype of NF1, and that the observed unilateral overgrowth of the left leg might be a rare consequence of the identified c.5727insT mutation.

Effect of numbers of sperm and timing of a single, post-cervical insemination on the fertility of weaned sows treated with OvuGel®.

Variability in estrus and ovulation requires multiple inseminations during estrus to ensure one AI occurs close to ovulation. Induction of ovulation after weaning improves synchrony of ovulation and allows for fixed time AI. However, the interaction between number of sperm in the AI dose and the timing of insemination has not been fully investigated. The objective of this study was to determine the effects of sperm numbers used in a single post-cervical insemination (PCAI) and the timing of insemination following induced ovulation in weaned sows. The experiment was performed using sows (n = 641) allotted by parity (1-6) and lactation length (19.5 d) to receive a single PCAI using 1.5 or 2.5 billion motile sperm at either 22, 26, or 30 h following administration of a GnRH agonist, triptorelin acetate (OvuGel®) at 96 h post-weaning. Sows received boar contact once daily 3-6 d following weaning. A sub-population of the sows (n = 499) were assessed for follicle size and ovulation utilizing ultrasound at 8 h intervals. There was no interaction of number of sperm and timing of insemination for any response measure (P > 0.10). Wean to estrus interval (4.8 d), duration of estrus (1.9 d), and expression of estrus (88.0%), were not different among treatments (P > 0.10). Of sows scanned by ultrasound at the time of OvuGel®, 88.2% had large follicles, 10.9% had small, medium or cystic sized follicles, and 0.9% had corpora lutea. The proportion of sows that ovulated averaged 94%, and differed by time of AI (P ≤ 0.05) but not by number of sperm. Pregnancy rate and farrowing rate tended to be affected by dose (P ≤ 0.10), while time of insemination affected pregnancy rate and tended to influence farrowing rate (P ≤ 0.10). Farrowing rate was greater (P < 0.0001) with use of 2.5 than 1.5 billion sperm and insemination at 22 and 26 h compared to 30 h after OvuGel® (P ≤ 0.10). Farrowing rate was also affected by parity, estrus expression, ovulation and ovarian abnormalities (P < 0.05). Of the 12% of weaned sows that did not exhibit estrus, approximately 50% farrowed a litter. Total born and born alive were affected by dose (P < 0.05) but not time of insemination with both measures increased with 2.5 compared to 1.5 billion sperm (P < 0.05). The results of this study indicate that induction of ovulation in weaned sows resulted in 88% of sows ovulating within a 24 h period. Fertility was improved with a single, fixed time AI using 2.5 compared to 1.5 billion motile sperm and insemination at 22-26 h after OvuGel® compared to 30 h.

Validation of Molecular Markers for Use With Adapted Sources of Fusarium Head Blight Resistance in Wheat.

Genetic control of resistance to Fusarium head blight (FHB) is quantitative, making phenotypic selection difficult. Genetic markers to resistance are helpful to select favorable genotypes. This study was conducted to determine if Fhb1 and Fhb5 present in the Sumai 3 source of FHB resistance occur in Sumai 3-derived North American spring wheat cultivars and to understand the appropriateness of using markers to select for the favorable alleles at these loci in breeding. Sumai 3-derived parents Alsen, ND3085, ND744, Carberry, and Glenn were used in crosses to generate 14 doubled haploid breeding populations. The parents and progeny were genotyped with five Fhb1 and three Fhb5 microsatellite markers. Progeny were selected based on performance relative to parents and other control cultivars in FHB nurseries near Portage la Prairie and Carman, MB. χ2 and t test analyses were performed on marker and FHB data. The χ2 test frequently determined the proportion of lines carrying molecular variants associated with FHB resistance increased following nursery selection for FHB. Similarly, the t test regularly demonstrated that selection for FHB resistance lowered the mean level of disease associated with resistant marker haplotypes. The study affirmed FHB resistance sources Alsen, Carberry, ND3085, and ND744 have Fhb1 and Fhb5 loci like Sumai 3, but no evidence was found that Glenn carries Fhb1 and Fhb5 resistance alleles. The results justified use of Fhb1 and Fhb5 markers for marker assisted selection in populations derived from Alsen, Carberry, ND3085, and ND744, but not Glenn. Combined or individual application of Xgwm493 and Xgwm533 in selection of genotypes carrying Fhb1, and Xgwm150, Xgwm304, and Xgwm595 for Fhb5 will enhance FHB resistance in wheat.

meso-Acetoxymethyl BODIPY dyes for photodynamic therapy: improved photostability of singlet oxygen photosensitizers.

We report two BODIPY based photosensitizers (Br2BOAc and I2BOAc) featuring an acetoxymethyl substituent at the meso-position. These photosensitizers show improved photostability against singlet oxygen, when compared to a BODIPY photosensitizer lacking the acetoxymethyl group. Both compounds were evaluated for photodynamic therapy against HeLa cells and photodynamic inactivation against E. coli bacteria. We show that the compounds readily embed in the lipid membranes of HeLa cervical cancer cells and efficiently induced light-dependent apoptosis at nanomolar concentration. Also, both compounds showed a substantial degree of photoinactivation of E. coli bacteria when used at low micromolar concentrations.

Potential to breed for mycorrhizal association in durum wheat.

The selection of genotypes under high soil fertility may alter the effectiveness of mycorrhizal symbioses naturally forming between crop plants and the mycorrhizal fungi residing in cultivated fields. We tested the hypothesis that the mycorrhizal symbiosis of 5 landraces functions better than the mycorrhizal symbiosis of 27 cultivars of durum wheat that were bred after the development of the fertilizer industry. We examined the development of mycorrhiza and the response of these genotypes to mycorrhiza formation after 4 weeks of growth under high and low soil fertility levels in the greenhouse. The durum wheat genotypes were seeded in an established extraradical hyphal network of Rhizophagus irregularis and in a control soil free of mycorrhizal fungi. The percentage of root length colonized by mycorrhizal fungi was lower in landraces (21%) than in cultivars (27%; P = 0.04) and in the most recent releases (29%; P = 0.02), which were selected under high soil fertility levels. Plant growth response to mycorrhiza varied from -36% to +19%. Overall, durum wheat plant breeding in Canada has increased the mycorrhizal development in wheat grown at a low soil fertility level. However, breeding had inconsistent effects on mycorrhizal development and has led to the production of cultivars with patterns of regulation ranging from unimproved to inefficient.

Effect of timing of relocation of replacement gilts from group pens to individual stalls before breeding on fertility and well-being.

Variation in gilt fertility is associated with increased replacement and reduced longevity. Stress before breeding is hypothesized to be involved in reduced fertility. This study tested the timing of gilt relocation from pens to individual stalls before breeding on fertility and well-being. The experiment was performed in replicates on a commercial research farm. After detection of first estrus, gilts ( = 563) were assigned to treatment for relocation into stalls 3 wk (REL3wk), 2 wk (REL2wk), or 1 wk (REL1wk) before breeding at second estrus. Subsets of gilts from each treatment ( = 60) were selected for assessment of follicles at second estrus. Data included interestrus interval, number of services, conception, farrowing, total born, and wean to service interval. Piglet birth weight was obtained on subsets of litters ( = 42/treatment). Measures of well-being included BW, backfat, BCS, lesions, and lameness from wk 1 after first estrus until wk 16. Gilt BW at wk 5 (158.4 kg) was not affected ( > 0.10) by treatment. Measures of BCS, lameness, and lesions at breeding and throughout gestation did not differ ( > 0.10). Treatment did not affect ( > 0.10) gilts expressing a normal interestrus interval of 18 to 24 d (83.4%) but did influence ( < 0.05) the proportion expressing shorter ( < 0.001) and longer ( < 0.001) intervals. Gilts in REL3wk had a shorter ( < 0.001) interestrus interval (20.7 d) than those in REL2wk and REL1wk (22.6 d). Gilts with shorter intervals ( = 24) had fewer total born while gilts expressing longer cycles ( = 65) had reduced farrowing rates. The number of services (1.9) and number of follicles (19.7) at breeding were not affected ( > 0.10) by relocation. There was no effect of treatment on farrowing rate (85.2%), born alive (12.6), or any litter birth weight measures ( > 0.10). The percentage of sows bred within 7 d after weaning (94.4%) was also not affected by treatment ( > 0.10). These results suggest that the timing of relocation before breeding had no effect on well-being or on the majority of gilts with normal estrous cycles and their subsequent fertility. However, a smaller proportion of the gilts exhibited shorter and longer interestrus intervals in response to relocation 1 or 3 wk before breeding. In cases where gilt fertility may be less than optimal, producers that relocate gilts from pens to stalls before breeding should evaluate interestrus interval as a response criterion.

Artificial insemination in pigs today.

Use of artificial insemination (AI) for breeding pigs has been instrumental for facilitating global improvements in fertility, genetics, labor, and herd health. The establishment of AI centers for management of boars and production of semen has allowed for selection of boars for fertility and sperm production using in vitro and in vivo measures. Today, boars can be managed for production of 20 to 40 traditional AI doses containing 2.5 to 3.0 billion motile sperm in 75 to 100 mL of extender or 40 to 60 doses with 1.5 to 2.0 billion sperm in similar or reduced volumes for use in cervical or intrauterine AI. Regardless of the sperm dose, in liquid form, extenders are designed to sustain sperm fertility for 3 to 7 days. On farm, AI is the predominant form for commercial sow breeding and relies on manual detection of estrus with sows receiving two cervical or two intrauterine inseminations of the traditional or low sperm doses on each day detected in standing estrus. New approaches for increasing rates of genetic improvement through use of AI are aimed at methods to continue to lower the number of sperm in an AI dose and reducing the number of inseminations through use of a single, fixed-time AI after ovulation induction. Both approaches allow greater selection pressure for economically important swine traits in the sires and help extend the genetic advantages through AI on to more production farms.

The Fertility of Frozen Boar Sperm When used for Artificial Insemination.

One of the limits to practical use of frozen boar sperm involves the lowered fertility when used for artificial insemination. Years of studies have shown that 5-6 billion sperm (approximately 3 billion viable) used in single or multiple inseminations results in pregnancy rates most often between 60 and 70% and with litter sizes between nine and 10 pigs. Yet today, it is not uncommon for studies to report pregnancy rates from 70 to 85% and litter sizes with 11-12 pigs. While global statements about the incidence and reasons for higher fertility are not conclusive, incremental fertility improvements appear independently associated with use of a minimum number of viable sperm (1-2 billion), insemination timing that increases the probability that sperm will be present close to ovulation for groups of females, selection for boar sperm survival following cryopreservation, and modification of the freeze and thaw conditions using additives to protect sperm from oxidative damage. Studies show that techniques such as intrauterine and deep uterine insemination can provide an opportunity to reduce sperm numbers and that control of time of ovulation in groups of females can reduce the need for multiple inseminations and improve the chance for AI close to ovulation. However, optimal and consistent fertility with cryopreserved boar sperm may require a multifaceted approach that includes boar selection and screening, strategic use of additives during the freezing and thawing process, post-thaw evaluation of sperm and adjustments in sperm numbers for AI, assessment of female fertility and ovulation induction for single insemination. These sequenced procedures should be developed and incorporated into a quality control system for improved fertility when using minimal numbers of cryopreserved boar sperm.

Genetic gain and economic values of selection strategies including semen traits in three- and four-way crossbreeding systems for swine production.

Four semen traits: volume (VOL), concentration (CON), progressive motility of spermatozoa (MOT), and abnormal spermatozoa (ABN) provide complementary information on boar fertility. Assessment of the impact of selection for semen traits is hindered by limited information on economic parameters. Objectives of this study were to estimate economic values for semen traits and to evaluate the genetic gain when these traits are incorporated into traditional selection strategies in a 3-tier system of swine production. Three-way (maternal nucleus lines A and B and paternal nucleus line C) and 4-way (additional paternal nucleus line D) crossbreeding schemes were compared. A novel population structure that accommodated selection for semen traits was developed. Three selection strategies were simulated. Selection Strategy I (baseline) encompassed selection for maternal traits: number of pigs born alive (NBA), litter birth weight (LBW), adjusted 21-d litter weight (A21), and number of pigs at 21 d (N21); and paternal traits: number of days to 113.5 kg (D113), backfat (BF), ADG, feed efficiency (FE), and carcass lean % (LEAN). Selection Strategy II included Strategy I and the number of usable semen doses per collection (DOSES), a function of the 4 semen traits. Selection Strategy III included Strategy I and the 4 semen traits individually. The estimated economic values of VOL, CON, MOT, ABN, and DOSES for 7 to 1 collections/wk ranged from $0.21 to $1.44/mL, $0.12 to $0.83/10 spermatozoa/mm, $0.61 to $12.66/%, -$0.53 to -$10.88/%, and $2.01 to $41.43/%, respectively. The decrease in the relative economic values of semen traits and DOSES with higher number of collections per wk was sharper between 1 and 2.33 collections/wk than between 2.33 and 7 collections/wk. The higher economic value of MOT and ABN relative to VOL and CON could be linked to the genetic variances and covariances of these traits. Average genetic gains for the maternal traits were comparable across strategies. Genetic gains for paternal traits, excluding semen traits, were greater in selection Strategy I than Strategies III and II. Genetic gains for paternal and maternal traits were greater in the 4- and 3-way schemes, respectively. The selection strategy including the 4 semen traits is recommended because this approach enables genetic gains for these traits without compromising the genetic gains for maternal traits and with minimal losses in genetic gains for paternal traits.

Enhanced fertility prediction of cryopreserved boar spermatozoa using novel sperm function assessment.

Due to reduced fertility, cryopreserved semen is seldom used for commercial porcine artificial insemination (AI). Predicting the fertility of individual frozen ejaculates for selection of higher quality semen prior to AI would increase overall success. Our objective was to test novel and traditional laboratory analyses to identify characteristics of cryopreserved spermatozoa that are related to boar fertility. Traditional post-thaw analyses of motility, viability, and acrosome integrity were performed on each ejaculate. In vitro fertilization, cleavage, and blastocyst development were also determined. Finally, spermatozoa-oviduct binding and competitive zona-binding assays were applied to assess sperm adhesion to these two matrices. Fertility of the same ejaculates subjected to laboratory assays was determined for each boar by multi-sire AI and defined as (i) the mean percentage of the litter sired and (ii) the mean number of piglets sired in each litter. Means of each laboratory evaluation were calculated for each boar and those values were applied to multiple linear regression analyses to determine which sperm traits could collectively estimate fertility in the simplest model. The regression model to predict the percent of litter sired by each boar was highly effective (p < 0.001, r(2) = 0.87) and included five traits; acrosome-compromised spermatozoa, percent live spermatozoa (0 and 60 min post-thaw), percent total motility, and the number of zona-bound spermatozoa. A second model to predict the number of piglets sired by boar was also effective (p < 0.05, r(2) = 0.57). These models indicate that the fertility of cryopreserved boar spermatozoa can be predicted effectively by including traditional and novel laboratory assays that consider functions of spermatozoa.

The effect of extender, method of thawing, and duration of storage on in vitro fertility measures of frozen-thawed boar sperm.

Frozen-thawed boar sperm (FTS) has reduced in vitro and in vivo life span compared to liquid semen. Experiments tested whether extenders, thawing procedures, and storage temperatures could extend the fertile life span of FTS. Experiment 1 tested the effect of six extenders on postthaw motility (MOT) and viability (VIA). Straws from boars (n = 6) were thawed, diluted into each extender, and evaluated at 20, 60, and 120 minutes. There was a trend (P = 0.08) for an extender-by-time interaction for MOT and effect of extender and time for MOT (P < 0.0001) and extender (P = 0.10) and time (P < 0.0001) for VIA. Experiment 2 evaluated the effect of temperature and time of thawing on in vitro fertility at intervals after thawing. Straws (0.5 mL) from different boar ejaculates (n = 15) were thawed at 50 °C for 10, 20, or 30 seconds or at 70 °C for 5, 10, or 20 seconds and evaluated at 5, 30, and 60 minutes. There was an effect of thawing treatment on MOT, VIA, and ACR (viable sperm with intact acrosomes, P < 0.0001) and an effect of time of evaluation (P < 0.0001) on MOT and ACR. Thawing at 70 °C for 20 seconds reduced (P < 0.05) MOT, VIA, and ACR compared to other treatments. Experiment 3 tested the effects of storage temperature and time after thawing using 20 ejaculates. Samples were thawed, diluted, and allotted to storage at 17 °C, 26 °C, or 37 °C with evaluation at 2, 6, 12, and 24 hours. There was a storage temperature and time effect and an interaction for MOT and VIA (P < 0.0001). Storage at 17 °C and 26 °C increased (P < 0.05) MOT over all times (38.5%) compared to 37 °C (26%), whereas MOT was reduced at intervals. Viability was also greatest with 17 °C and 26 °C compared to 37 °C and was also affected by time and decreased with time. These results indicate that FTS can be held at 17 °C or 26 °C for up to 2 hours before use and would allow for preparation of multiple doses. These data suggest in vitro fertility of FTS is affected by extenders, thawing, and storage.

Stripe rust and leaf rust resistance QTL mapping, epistatic interactions, and co-localization with stem rust resistance loci in spring wheat evaluated over three continents.

KEY MESSAGE: In wheat, advantageous gene-rich or pleiotropic regions for stripe, leaf, and stem rust and epistatic interactions between rust resistance loci should be accounted for in plant breeding strategies. Leaf rust (Puccinia triticina Eriks.) and stripe rust (Puccinia striiformis f. tritici Eriks) contribute to major production losses in many regions worldwide. The objectives of this research were to identify and study epistatic interactions of quantitative trait loci (QTL) for stripe and leaf rust resistance in a doubled haploid (DH) population derived from the cross of Canadian wheat cultivars, AC Cadillac and Carberry. The relationship of leaf and stripe rust resistance QTL that co-located with stem rust resistance QTL previously mapped in this population was also investigated. The Carberry/AC Cadillac population was genotyped with DArT(®) and simple sequence repeat markers. The parents and population were phenotyped for stripe rust severity and infection response in field rust nurseries in Kenya (Njoro), Canada (Swift Current), and New Zealand (Lincoln); and for leaf rust severity and infection response in field nurseries in Canada (Swift Current) and New Zealand (Lincoln). AC Cadillac was a source of stripe rust resistance QTL on chromosomes 2A, 2B, 3A, 3B, 5B, and 7B; and Carberry was a source of resistance on chromosomes 2B, 4B, and 7A. AC Cadillac contributed QTL for resistance to leaf rust on chromosome 2A and Carberry contributed QTL on chromosomes 2B and 4B. Stripe rust resistance QTL co-localized with previously reported stem rust resistance QTL on 2B, 3B, and 7B, while leaf rust resistance QTL co-localized with 4B stem rust resistance QTL. Several epistatic interactions were identified both for stripe and leaf rust resistance QTL. We have identified useful combinations of genetic loci with main and epistatic effects. Multiple disease resistance regions identified on chromosomes 2A, 2B, 3B, 4B, 5B, and 7B are prime candidates for further investigation and validation of their broad resistance.

Effects of altering the dose and timing of triptorelin when given as an intravaginal gel for advancing and synchronizing ovulation in weaned sows.

Previous studies have shown that triptorelin gel (TG) given intravaginally in gel form is effective for advancing the time of ovulation in weaned sows. Three experiments were performed to determine the effects of altering the dose and timing of administration of intravaginal TG for advancing and synchronizing ovulation in weaned sows. In all experiments, estrus was detected twice or three times daily and ultrasound was performed to determine ovulation at 8-hour intervals. In experiment 1, sows (n = 131) received intravaginal gel containing 0 (Placebo), 25, 100, or 200 µg of TG at 96 hours after weaning and sows were inseminated on each day of standing estrus. Wean-to-estrus interval and duration of estrus were correlated (P < 0.0001) with estrus duration longer in TG (P < 0.05) compared with Placebo. More sows ovulated (P < 0.001) by 48 hours after treatment with 200 (81%), 100 (64%), and 25 µg (63%) of TG compared with Placebo (42%). The farrowing rate and total pigs born did not differ (P > 0.10). In experiment 2, sows (n = 126) received 200 µg of TG at 72, 84, or 96 hours after weaning or were untreated (Control-96). Sows receiving TG were inseminated once 24 to 28 hours after treatment. Control-96 sows were inseminated on each day of standing estrus. Wean-to-estrus interval was not affected by treatment, but wean-to-ovulation interval was reduced (P < 0.05) by TG-72 and TG-84 compared with TG-96 and Control-96. More sows ovulated 40 hours after treatment (P < 0.001) with TG-72 (56.5%) and TG-84 (32.2%) compared with TG-96 and Control-96 (13%) and for all TG treatments 48 hours after treatment (64%) compared with Control-96 (34%, P < 0.05). The farrowing rate was lower (P < 0.05) for sows assigned to TG-72 and TG-84 compared with TG-96 and Control-96, whereas the number of liveborn pigs did not differ (P > 0.10). In experiment 3, sows (n = 113) were assigned to receive no treatment (Control), intravaginal gel alone (Placebo), or 200 µg of TG given intravaginally (OvuGel) at 96 hours after weaning. Wean-to-estrus interval did not differ, but the duration of estrus tended (P < 0.10) to be reduced with OvuGel compared with the other treatments. More sows ovulated (P < 0.001) by 48 hours after OvuGel treatment (79.1%) compared with Control (46.4%) and Placebo (37.9%) and by 56 hours (P < 0.05). The farrowing rate and the number of liveborn pigs did not differ among treatments. The results of these studies indicate that 200 µg of TG given intravaginally at 96 hours after weaning (OvuGel) synchronizes ovulation and results in fertility similar to Controls.