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The endogenous steroid hormone ouabain induces capacitation of bull sperm acting through its receptor Na + /K + -ATPase on the sperm plasma membrane. Progesterone (P4) is believed to act through the sperm membrane P4 receptor (mPR) to induce non-genomic signalling leading to capacitation and/or acrosome reaction (AR) in the sperm of some species, but the exact nature of this receptor molecule on bull sperm is not known. In amphibian oocytes, P4 acts through the low-affinity ouabain binding site on Na + /K + -ATPase to induce signalling highly reminiscent of ouabain's signalling that initiates capacitation. This study hypothesized that ouabain and P4 interact agonistically or antagonistically to induce bull sperm capacitation. Sperm were incubated with 0, 12.5, 25, 50 and 100 μM ouabain, P4 or ouabain + P4 (12.5, 25 and 50 μM each) under capacitating conditions, and capacitation was assessed microscopically looking at the acrosome status of sperm and as the amount of protein tyrosine phosphorylation (Tyr-P). Both steroids stimulated Tyr-P of certain sperm proteins, but ouabain caused tyrosine phosphorylation of more proteins than P4 and stimulated significantly more overall Tyr-P (P < 0.05). Ouabain also was the only steroid to stimulate significant microscopically-evident AR. When sperm were co-incubated with the two steroids, P4 partially inhibited ouabain-induced Tyr-P and AR. These results suggest that P4 and ouabain may both interact with Na + /K + -ATPase, but ouabain is the more effective hormone. Ouabain may, therefore, be the primary physiological inducer of bovine capacitation., (Copyright © 2018. Published by Elsevier Inc.)

Although sperm and seminal plasma differ in their origin, biophysical and biochemical properties of seminal plasma influence the sperm function. Seminal plasma is a fluid medium containing substances from testes, epididymides and accessory glands. Composition of seminal plasma varies among animal species and in boars, prostate and bulbourethral glands are major contributors to the volume and contents. While the origin of some components of seminal plasma are known, the source of recently discovered seminal plasma microRNAs remains unknown, in part due to the difficulty of recovering and characterizing RNA from porcine sperm and seminal plasma. To test the hypothesis that seminal plasma miRNAs interact with sperm, the first objective was to validate protocols for recovering RNAs from porcine seminal plasma and sperm, whereas the second objective was to characterize expression patterns of 84 prioritized microRNAs employing real time PCR methodology. The study identified a relationship between sperm and seminal plasma microRNAs, based on the normalized threshold cycle of amplifying cDNA in sperm and seminal plasma from the same semen of Landrace boars. Therefore, it was concluded that seminal plasma miRNAs may originate from sperm or these miRNAs may shuttle between sperm and seminal plasma in order to facilitate cell-to-cell communication., (Copyright © 2018 Elsevier Inc. All rights reserved.)

This is the first study where the systematic application of theories and techniques used in mammalian sperm cryopreservation have been applied to honey bee (Apis mellifera L.) semen as a means to improve postthaw viability of cryopreserved sperm. Six newly designed diluents, three cryoprotectants (dimethyl sulfoxide, DMA, glycerol), and five diluent:semen ratios (1:1, 3:1, 6:1, 9:1, and 12:1) were tested. In addition, the sperm freezing tolerance of three honey bee strains was evaluated. Specific protocols were designed to control semen freezing and thawing rates. Sperm motility was assessed visually, whereas sperm viability was assessed using SYBR-14 and propidium iodide fluorescent stains. Diluent treatments did not affect fresh (nonfrozen) sperm viability yet affected fresh sperm motility (P<0.05). Based on these assessments, two diluents were chosen and used in all successive cryopreservation experiments. Using the selected diluents, semen was collected at various diluent:semen ratios, along with one of the three cryoprotectants. Semen collected at high dilution ratios, using a hypotonic antioxidant diluent containing catalase, in combination with dimethyl sulfoxide, provided higher postthaw sperm viability than that of all other combinations tested (68.3+/-5.4%; P<0.05). Using this combination of dilution ratio, diluent, and cryoprotectant, there were no differences among honey bee strains for postthaw sperm viability (P=0.805). Nevertheless, these new semen dilution and freezing methods improved postthaw viability of sperm to levels that could theoretically sustain worker populations in colonies, thus providing potential for further optimization of cryopreservation techniques for the genetic preservation and improvement of honey bee genotypes.

Sperm-mediated DNA transfer can be used to transfer exogenous DNA into the oocyte for the production of transgenic animals. In spite of controversy in the literature, sperm-mediated DNA transfer is a simple and quick technique that can be used in routine breeding programs (AI, embryo transfer and IVF). The main objective of this study was to determine the factors affecting the spontaneous uptake of exogenous DNA by bull spermatozoa. For this purpose, fresh and frozen spermatozoa (0.25 x 10(6)), from the same ejaculate from each of four bulls were co-incubated with fluorescent-labeled green fluorescent protein (GFP) and chloremphenicol acetyltransferase (CAT) plasmids at 37 degrees C for 30 min. Neither bull nor plasmid significantly affected the uptake of exogenous DNA. However, transfection efficiency was higher in frozen-thawed versus fresh spermatozoa (P<0.001). Regardless of whether transfected spermatozoa were alive or dead, all transfected spermatozoa were immotile. It can be concluded that a population of spermatozoa is present in bull semen which has the ability to uptake exogenous DNA spontaneously. There is tremendous scope to improve transfection efficiency of spermatozoa while maintaining motility; this needs to be achieved in order to more easily use this technique in transgenesis. However, live-transfected bull spermatozoa clearly can incorporate exogenous DNA and should be usable in intracytoplasmic sperm injection protocols.

Boars in an artificial insemination centre have been selected for their superior genetic potential, with 'superior' being defined as having traits the customer wants transmitted to his herd. The ability to meet the customers' needs depends on the heritability of the trait, the geneticist's success in devising a selection scheme for the trait in balance with other economically important traits, and the boar's ability to produce sperm that can fertilise oocytes. Genetic evaluation research over the past 20 years has greatly increased the number of traits for which a boar can be selected: currently in the Canadian national program, these include age at 100 kg, backfat at 100 kg, feed efficiency, lean yield and litter size. In the near future, traits that are very likely to be added to this selection list include piglet survival, marbling, loin eye area and structure traits. In Canada, sires are ranked on two estimated breeding value (EBV) indices; one, focused on development of terminal sire lines, is based on the growth and yield traits and another, primarily focused on maternal line development, de-emphasises these traits and incorporates litter size. Boars that are in Canadian AI centres because of their excellent growth traits are typically in the top 5-10% of the national population for terminal sire line index, but they may be only average or substandard for litter size. Conversely, boars selected to be in the top 5-10% for conveying such reproductive traits as litter size may only be in the top 33% for growth traits. The more offspring from a superior boar in either of these indices, the faster the population average for the trait improves. The original sire gets knocked out of the elite group, is culled and replaced by a higher ranked young boar from the now improved general population. Although genetic superiority should govern an AI centre's selection and culling of boars, decision-making in real life is seldom that simple. Selection criteria may be contradictory as above, or a boar with truly superior traits may be excluded because a newly-developed molecular genetics test determines he carries an undesirable gene such as PSS, RN or others being developed. Selection for terminal sire or maternal line traits can ignore important practical factors that affect an AI centre--boars with superior genetics may not produce good semen because skeletal or penile problems prevent ejaculation, or because sperm production is poor due to a genetic flaw, disease, or some other cause. Interestingly, selection pressure for one trait may inadvertently select for a trait that is linked but whose linkage is unrecognised, and such unintentionally selected genes could benefit, harm, or have no effect on production traits. An AI centre serving a variety of customers must select boars in anticipation of their customers' needs (including new, foreign and niche markets). A centre should also review its genetic evaluation results and progeny records, both to critique its own selection success and to try to detect unexpected linkages. Finally, an AI centre needs to predict its own future, selecting not just for production traits for the swine producer, but also for factors that enhance the centre's efficiency including boar conformation and temperament, and sperm quantity, quality and hardiness. Can we select for efficiency? Our colleagues in dairy cattle AI evaluate bull performance--should the swine industry consider evaluation of male fertility traits?