Lectins are glycoproteins of a non-immune origin often used as histochemical reagents to study the distribution of glycoconjugates in different types of tissues. In this study, we performed a comparative cellular localization of sugar residues in bull and donkey testes using immunofluorescent lectin histochemistry. We inspected the cellular localization of the glycoconjugates within the testes using 11 biotin-labeled lectins (LCA, ConA, PNA, WGA, DBA, SBA, ECA, BPL, PTL-II, UEA-1, and PHA-E4) classified under six groups. Although the basic testicular structure in both species was similar, the cellular components showed different lectin localization patterns. The statistical analysis revealed no significant association between the intensity of labeling and different variables, including group and type of lectin and type of cell examined, at p < 0.05. However, a stronger response tended to occur in the donkey than in the bull testes (odds ratio: 1.3). These findings may be associated with the different cellular compositions of the glycoproteins and modification changes during spermatogenesis. Moreover, glycoconjugate profiling through lectin histochemistry can characterize some cell-type selective markers that will be helpful in studying bull and donkey spermatogenesis.

Selecting bulls for reproductive soundness requires that the bull be structurally sound, free of abnormalities that impair his ability to produce adequate numbers of motile, morphologically normal spermatozoa, and be able to successfully complete coitus. This review discusses the diagnosis and etiology of abnormalities of the penis, prepuce as well as common musculoskeletal conditions that prevent normal pasture breeding soundness. A review of testicular and thermoregulation addresses the potential impact of musculoskeletal disorders on normal sperm production.

Up to the 18th century, the prevailing view of reproduction, or ‘generation’ as it was referred to, was that organisms develop from miniatures of themselves, termed preformation. The alternative theory, epigenesis, proposed that the structure of an animal emerges gradually from a relatively formless egg. The teachings of the Ancient Greeks who argued either that both sexes each contributed ‘semen’ to form the embryo, or held a more male-centred view that the female merely provided fertile ground for the male seed to grow, dominated thinking until the 17th century, when the combined work of numerous scholars led to the theory that all female organisms, including humans, produced eggs. The sequence of events leading to the commercial use of artificial insemination (AI) date back to the discovery of sperm in 1678, although it took almost 100 years to demonstrate that sperm were the agents of fertilisation and a further 100 years for the detailed events associated with fertilisation to be elucidated. The first successful AI, carried out in the dog, dates back to 1780 while it was not until the early to mid-1900s that practical methods for AI were described in Russia. Inspired by the Russian success, the first AI cooperative was established in Denmark in 1936 and later in the United States in 1938. The next major advances involved development of semen extenders, addition of antibiotics to semen, and the discovery in 1949 that glycerol protected sperm during cryopreservation. Almost four decades later, the flow cytometric separation of X- and Y-bearing sperm opened a new chapter in the application of AI for cattle breeding. As we look forward today, developments in imaging sperm and breakthroughs in gene editing and stem cell technology are opening up new possibilities to manipulate reproduction in a way never thought possible by the pioneers of the past. This review highlights some of the main milestones and individuals in the history of sperm biology and the development of technologies associated with AI in cattle.

This review focuses on current understanding of prenatal, prepubertal and post-pubertal development of the male reproductive system of cattle. The critical developmental events occur during the first 3 to 4 months of gestation and the first ~6 to 9 months after birth. The Wilms Tumor-1 and SRY proteins play critical roles in early development and differentiation of the fetal testis, which in turn drives gestational development of the entire male reproductive system. The hypothalamic–pituitary–gonadal axis matures earlier in the bovine fetus than other domestic species with descent of the testes into the scrotum occurring around the 4th month of gestation. An array of congenital abnormalities affecting the reproductive system of bulls has been reported and most are considered to be heritable, although the mode of inheritance in most cases has not been fully defined. Early postnatal detection of most of these abnormalities is problematic as clinical signs are generally not expressed until after puberty. Development of genomic markers for these abnormalities would enable early culling of affected calves in seedstock herds. The postnatal early sustained increase in lutenising hormone secretion cues the rapid growth of the testes in the bull calf leading to the onset of puberty. There is good evidence that both genetic and environmental factors, in particular postnatal nutrition, control or influence development and maturation of the reproductive system. For example, in Bos taurus genotypes which have had sustained genetic selection pressure applied for fertility, and where young bulls are managed on a moderate to high plane of nutrition puberty typically occurs at 8 to 12 months of age. However, in many Bos indicus genotypes where there has been little selection pressure for fertility and where young bulls are reared on a low plane of nutrition, puberty typically occurs between 15 to 17 months. Our understanding of the control and expression of sexual behavior in bulls is limited, particularly in B. indicus genotypes.

Knowledge of the capabilities and limitations of the reproductive capacity of bulls is vital in maximizing reproductive efficiencies. Bull semen collection guidelines established by researchers and industry personnel to maximize the sperm harvest from bulls have been evolving for more than 60 years. Today, a mature artificial insemination industry employs those strategies to meet demands. These efficient management schemes exploit the reproductive potential of each sire while minimizing the associated risk of injury to bulls and reduce associated production costs. Personnel employed by a semen producing facility must be authorized to make effective and rational decisions based on principles of bull sexual behavior and reproductive physiology. Furthermore, collection facilities must be well planned to allow for the safe presentation of novel sexual situations while affording maximum safety for employees and proper footing for bulls. Normal bulls produce and ejaculate tremendous numbers of sperm. Most bulls have a sufficient libido for routine sexual activity, but become satiated to predictable stimulus situations. Frequent changes to the novelty should allow weekly harvest of four to six ejaculates per week for most bulls. Utilizing the physiological characteristics associated with each ejaculate to establish the collection frequency of each bull, and empowering an integrated collection and laboratory staff to monitor and make adjustments to the ejaculation frequency are necessary in maximizing the sperm harvest. Young bulls can ejaculate 10 to 20 billion sperm per week, and mature bulls should ejaculate 40 to 60 billion sperm per week. Semen collection management procedures should be reviewed when bulls do not meet production goals.

The aim of this work was to examine the influence of cryostorage duration of Pinzgau bull's insemination doses (IDs) on some sperm traits. The IDs were frozen by a slow freezing method and stored in liquid nitrogen for different periods: less than 8 years (group 1), 8–13 years (group 2) and 14–18 years (group 3). Motility (CASA), pathological sperm rate (Giemsa staining), apoptotic (Yo-Pro-1-positive) and necrotic (propidium iodide-positive) cell occurrence and fertilizing ability (penetration/fertilization test) of spermatozoa were evaluated post-thaw. The average post-thaw sperm motility in all examined groups was over 40%. No significant influence of storage length either on the sperm total motility or progressive movement was revealed. In each tested group the average rate of malformed spermatozoa did not exceed 20%. No effect of cryostorage length on the occurrence of apoptotic or necrotic sperm was noted. Similarly, penetrating/fertilizing ability of sperm did not differ among the groups, excepting differences in the rate of pronuclei (PN) formation. In group 1, 72.9% of eggs showed two visible PN following 20 h incubation with sperm, whilst in groups 2 and 3 only 67 and 54.5% of zygotes, respectively, had both PN at this time. These results revealed no influence of storage time on the bull spermatozoa in all parameters excepting the rate of PN formation. As high inter-male variability was observed in the susceptibility of bull sperm to cryostorage, individual differences should be taken into account when semen from individual bulls is to be stored for a long time.

Recent research reveals that in the so-called Mithraic tauroctony, the god is, in fact, wounding a bull, not killing it. I argue that the scene combines the overall design of evil-eye amulets with the pose of the goddess Nike performing a military sphagion and I suggest that the scene must have been understood by its creator and by some viewers, at least, to offer protective power in this world, as well as salvific assurance about the next, a dual focus that seems to have been especially strong in Mithraism.

To date, there has been little improvement in cryopreservation of bull sperm due to lack of understanding of the freezing mechanisms. Therefore, this study set out to investigate expression levels of fertility-associated proteins in bull sperm, and in particular the relationship between the 90 kDa heat-shock protein (HSP90) and the sperm characteristics after freezing–thawing. Semen was collected from eight Holstein bulls by artificial vagina. Characteristics of these fresh semen, including sperm motility, morphology, viability and concentration, were evaluated. Sperm quality was also assessed after freezing–thawing. Eight ejaculates were divided into two groups based on freezing resistance and sperm motility. Sperm proteins were extracted and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and western blotting were performed. SDS-PAGE results showed that there was substantial diversity in 90 kDa proteins in the frozen–thawed sperm and HSP90 was confirmed as one of the 90 kDa proteins by western blot. This study indicated that HSP90 expression correlated positively with sperm quality. The amount of expressed 90 kDa proteins in the high freezing resistance (HFR) group was significantly higher than that in the low freezing resistance (LFR) group (P < 0.05). Thus, higher expression of HSP90 could probably lead to the higher motility and freezing resistance of sperm found after freezing–thawing. Therefore, we concluded that level of HSP90 expression could be used to predict reliably and simply the freezing resistance of bull sperm.

The bovine's long generation interval results in a delay of several years when evaluating mammary specific transgenes in genetically engineered animals. This experiment was conducted to evaluate the feasibility of reducing that waiting period. Lactation was induced in prepubertal bull and heifer calves as a means of predicting transgene behaviour during subsequent post-parturient lactations in the heifers themselves, and in daughters sired by the bulls. The animals carry a lactation-specific transgene encoding lysostaphin, an antimicrobial protein that kills Staphlococcus aureus, a mastitis-causing pathogen. Oestrogen, progesterone and dexamethasone were administered as previously described (Ball et al. 2000) to nine heifers (five transgenics) ranging in weight from 80 to 145 kg. Eight bull calves (seven transgenics) weighing 81–178 kg received additional oestrogen and progesterone injection prior to dexamethasone treatment. All nine heifers responded to the milk induction scheme yielding between 19 ml and 4·5 l over 5 d. Milk volume from the four responding males (30 μl to 2·5 ml) was significantly less than that harvested from females (P=0·025). Only bull calves >117 kg had a positive response. Lysostaphin was detected in all transgenic prepubertal heifers and in two transgenic prepubertal bull calves induced. A positive relationship was observed between lysostaphin's stapholytic activity in the two types of lactations (r2=0·907, P<0·001) thus providing a useful means of predicting subsequent lysostaphin production in post-partum milk.

This study was conducted to investigate the possibility of using bovine oocytes for a heterologous fertility test by intracytoplasmic sperm injection (ICSI) and to compare the pronuclear formation of ram, bull and minke whale spermatozoa after injection into bovine oocytes. Bovine oocytes were cultured in vitro for 24 h and those with a polar body were selected for ICSI. Frozen-thawed semen from the three species were treated with 5 mM dithiothreitol for 1 h and spermatozoa were killed by storing them in a -20 °C refrigerator before use. ICSI was performed using a Piezo system. Three experiments were designed. In experiment 1, a higher (p < 0.05) male pronuclear formation rate was found in the oocytes injected with ram (52.6%) or bull (53.4%) spermatozoa than with minke whale spermatozoa (39.1%). In experiment 2, sperm head decondensation was detected at 2 h after ICSI in the oocytes injected with a spermatozoon of each species. Male pronuclei were first observed at 4 h in the oocytes injected with ram or bull spermatozoa and at 6 h in oocytes injected with minke whale spermatozoa. The mean diameters of male pronuclei derived from both whale and bull spermatozoa were larger than those from ram spermatozoa (30.4 μm and 28.3 μm vs 22.4 μm, p < 0.005). The mean diameter of female pronuclei in the oocytes injected with whale spermatozoa was also larger than with ram spermatozoa (29.3 μm vs 24.7 μm, p < 0.05). The development of male and female pronuclei was synchronous. In experiment 3, ethanol-activated oocytes injected with a spermatozoon from any of the three species achieved significantly higher (p < 0.05-0.001) cleavage rates than control oocytes. Blastocyst formation was only observed when bull spermatozoa were used. The results of this study indicate that dead foreign spermatozoa can participate in fertilisation activities in bovine oocytes after ICSI.