Prevalence of chlamydiae in boars and semen used for artificial insemination

Although there are indications for venereal transmission of chlamydiae in pigs, direct diagnostic evidence on the presence of these bacteria in boars and boar semen in particular is still incomplete. We investigated boars from two studs (A, B) in semen (A: n = 174; B: n = 100) and faeces (A: n = 174; B: n = 24) for chlamydiae using ompA-PCR and partial ompA gene sequencing. Additionally, blood serum was examined for chlamydial antibodies using an indirect ELISA (A: n = 171; B: n = 62). Chlamydiae were found in 9 (5.2%) and 24 (24.0%) semen specimens, and in 71 (40.1%) and 2 (8.3%) faecal samples from boars of stud A and B, respectively. Regarding individual chlamydial species, Chlamydophila psittaci and Chlamydia suis were identified most frequently, with the former predominating in semen (in 23 out of 33 positive samples) and the latter in faeces (68/73). In contrast, Chlamydophila pecorum was found only sporadically. Chlamydial antibodies were detected in 80 (46.8%) and 6 (9.7%) boars of stud A and B, respectively. No correlation was observed between the data from serology and PCR of semen or faeces in either of the studs. In conclusion, detection of chlamydiae in semen of boars suggests a potential for venereal transmission. Whether the high overall prevalence of chlamydial infections reflects a general situation in boars needs to be investigated. Serological testing failed to identify boars shedding chlamydiae in their semen.     

Prevalence of chlamydiae in semen and genital tracts of bulls, rams and bucks

Chlamydiae infect male genital organs of ruminants. However, little is known about their prevalence. Hence, we investigated fresh and cryopreserved semen (bulls: n=304; rams: n=78; bucks: n=44) by polymerase chain reaction (PCR), as well as genital organs (bulls: n=13; rams: n=10; bucks: n=6) by immunohistochemistry (IHC) and PCR. Sera from bulls (n=104) and small ruminants (n=61) were tested by LPS and rMOMP (recombinant major outer membrane protein) ELISA and competitive ELISA (cELISA), respectively. Three PCR assays were compared in this study for detection of chlamydial DNA in semen: 16S rRNA, IGS-S (intergenic spacer 16S/23S-short), and IGS-L (intergenic spacer 16S/23S-long) PCRs. PCR sensitivity and inhibitory effects were determined by spiking semen with Chlamydophila (Cp.) abortus DNA. In bull semen, detection limits of the 16S, IGS-S and IGS-L PCRs were 10, 10, 100 templates, respectively. However, PCR sensitivity was reduced in ram and buck semen suggesting the presence of potential PCR inhibitors. Of 304 bull semen samples, the 16S PCR revealed DNA of chlamydiae in 20 samples (6.6%), including Cp. abortus (n=2), Cp. psittaci (n=1), Chlamydia suis (n=2), and Chlamydia-like organisms (n=15). In rams, one semen sample was positive for Chlamydia-like organism. All investigated male genital organs were negative for Chlamydia. Serology revealed 47.1% (49/104) positive bulls by LPS ELISA. Of these, 30 samples were positive by rMOMP ELISA, predominantly for Cp. pecorum. In small ruminants, cELISA displayed 34.8% (16/46) and 60% (9/15) positivity for Cp. abortus in rams and bucks, respectively. There was no correlation between serology and PCR of semen. The presence of chlamydiae in semen suggests the possibility of venereal transmission, although risk may be low in Switzerland.     

Investigation of Chlamydiaceae in semen and cauda epididymidis and seroprevalence of Chlamydophila abortus in breeding bulls

Background

Reproductive disorders associated with chlamydial infection have been reported worldwide in cattle and there are indications of potential venereal transmission.

Methods

Semen samples from 21 dairy bulls and cauda epididymidis tissue samples from 43 beef bulls were analysed for chlamydial agent by real-time polymerase chain reaction (PCR) including an internal amplification control (mimic). Additionally, presence of antibodies against Chlamydophila (Cp.) abortus among the bulls was investigated with the commercial Pourquier^® ELISA Cp. abortus serum verification kit.

Results

No chlamydial agent was detected by PCR in either the semen samples or in the tissue samples. Additionally, no antibodies against Cp. abortus were detected.

Conclusions

The results suggest that Cp. abortus is very rare, or absent in Swedish bulls and thus the risk for venereal transmission of chlamydial infection through their semen is low. However, because Chlamydophila spp. infection rates seem to differ throughout the world, it is essential to clarify the relative importance of transmission of the infection through semen on cattle fertility.     

Chlamydiae in oviducts and uteri of repeat breeder pigs

Chlamydial infections of the genital organs cause reproductive failure in female pigs, and the uterus is recognized a target tissue for an infection. In contrast, information on the effect of chlamydiae on the porcine oviduct is patchily and inconclusive, although the bacteria are known to cause severe tubal defects in humans and laboratory animals. The aim of this study was to examine the segments ampulla (A), isthmus (I) and utero-tubal junction of the left (n=20) or both (n=22) oviducts, and uteri (U) from 42 culled repeat breeder pigs for chlamydiae using ompA-PCR, partial ompA gene sequencing, immunohistochemistry (IHC) and microscopy of tissue specimens for histopathology. As revealed by PCR, among a total of 26 chlamydia-positive females, 19 were tested positive in one or more segments of one or both oviducts, 14 were found positive in the uterus, and concomitant infections of both organs were observed in 7 of them. Sequencing of 33 PCR products revealed the following chlamydial species: Chlamydophila (Cp.) psittaci (n=18), Cp. abortus (n=2), Chlamydia (C.) suis (n=10), and C. trachomatis (n=3). Immunopositive staining was observed within the surface epithelium (in A, I, U), stromal tissue (in I, U) and muscular layer (in A, I, U). A total of 24 females had inflamed oviductal segments (in A and/or I) and 36 inflamed uteri. However, there was no relationship between histopathology and results of PCR or IHC. In conclusion, chlamydiae were found to infect oviducts and uteri of pigs. Further studies are required to clarify whether chlamydial infection causes specific histopathology and alters tubal function.     

Non-infectivity of semen from bulls infected with bovine leukosis virus

Semen and serum were obtained from four bovine leukosis virus (BLV) infected bulls from each of eight bull studs. The samples from the 32 bulls were frozen and stored in liquid nitrogen for subsequent testing. The sera were tested for antibodies to BLV by the agar gel immunodiffusion (AGID) method. Thirty of the bulls were found to be infected with BLV. Pairs of sheep were intraperitoneally inoculated with semen pools of the four bulls from each bull stud. None of the sheep developed antibodies to BLV. A later challenge with BLV infected lymphocytes resulted in the infection of all challenged sheep indicating that they were susceptible to BLV infection. The results provide evidence that transmission of BLV via leukocyte free semen from BLV infected bulls does not occur.     

Flow of blood to the ovaries of ewes throughout the estrous cycle: Niswender,G.D., R.T. Moore,A.M. Akbar,T.M. Nett and M.A. Diekman: Biol. Repr., 13: 381, 1975. Department of Physiology and Biophysics,Colorado State University,Fort Collins,Colorado 80523

This study was undertaken to determine whether a specific antimycoplasmal immune response could be detected in the male bovine genital tract and to better define mechanisms of immunity at that site. Specific $\text{Mycoplasma agalactiae}$ subsp. $\text{bovis}$ agglutinins were titrated in the serum, semen and preputial mucus extracts of two bulls with $\text{M. agalactiae}$ induced chronic seminal vesiculitis and of one normal control bull. Titers from infected bulls averaged 64 for serum, 1024 for semen and <8 for preputial mucus extracts whereas the control bull titers were 16 for serum, <8 for semen, and <8 for preputial mucus extracts. Because of the high semen agglutinin titers from infected bulls it was proposed that semen titers may be more useful diagnostically than serum titers.Studies of immunoglobulin levels in semen revealed that IgA, IgG₁ and IgG₂ levels were all much higher in infected bulls than in the control bull. These high semen IgA levels together with the high semen agglutinin titers indicated a local secretory immune response in genital tracts of infected bulls.     

Infectious Bovine Rhinotracheitis Virus Infection in Bulls, with Special Reference to Preputial Infection

In an experiment with infectious bovine rhinotracheitis (IBR) virus in two bulls, observed over a period of 122 weeks, the pattern of virus release was studied. Recurrent, unprovoked release of virus was demonstrated after one year in a nasal washing from one of the bulls and in preputial washings of both on 13 and 4 occasions, respectively, and finally in weeks 113 and 110, although clinical disease was not observed. During periods of recurrent virus release, concentrations of virus in the prepuce were generally much lower than during the period of primary infection; usually, however, they were not of negligible titer. The frequency of such periods and the virus titers observed strongly suggest that an IBR antibody carrier should always be considered as a potential source of infection to other animals. When virus was demonstrated in semen an almost equal amount was found in the preputial washing (50 ml). In week 120, virus replication in the respiratory tract and prepuce was induced in both bulls by prednisolone injections. It is concluded that antibody carriers will rarely attain a state of absolute immunity.     

Relationship between sperm-zona pellucida binding assays and the 56-day nonreturn rate of cattle inseminated with frozen-thawed bull semen

Assays based on sperm-zona pellucida binding have been developed as diagnostic tests to predict the fertilizing potential of mammalian spermatozoa. Recently, we reported on the development of a sperm-zona pellucida binding assay (SZBA) for bull spermatozoa. The aim of the present study was to develop a hemi-zona assay (HZA) for bull spermatozoa and to investigate the relationship between SZBA and HZA outcomes and in vivo fertility. Frozenthawed semen samples from 8 fertile Swedish Red and White bulls (one ejaculate per bull) designated as the test semen samples and a single ejaculate from a fertile Holstein-Friesian bull designated as the control semen sample were used in this study. In the SZBA, 2 groups of 20 oocytes per semen test sample and in the HZA a minimum of 6 matching pairs of hemizonae were used for comparison of sperm binding with control semen. Sperm binding to matching hemi-zonae of individual semen samples was equal, and clearly demonstrated the feasibility of the HZA for cattle. A significant correlation was found between the SZBA and the HZA indices obtained from the different semen test samples (r = 0.42, P < 0.001; n = 67). There was no significant relation between the SZBA indices and the 56-d nonreturn rate of the test samples. However, the HZA indices of the semen test samples and the 56-d nonreturn rate were significantly correlated (r = 0.46, P < 0.0001; n = 67). It is concluded that HZA can be regarded as a potential assay for predicting the fertilizing ability of bovine semen samples. However, further studies using more semen samples are necessary to confirm this view.     

Semen and reproductive profiles of genetically identical cloned bulls

In this comparative study, reproductive parameters and semen characteristics of cloned bulls (n = 3) derived from somatic cell nuclear transfer (SCNT) were compared to their original cell donor Holstein-Friesian (n = 2) bulls from the same enterprise to assess the differences in reproductive potential between a donor bull and its clones. The parameters evaluated included motility of fresh, frozen-thawed and Percoll-treated frozen-thawed spermatozoa, as well as in vitro fertilization (IVF) ability, embryo quality, birth and survival of calves following IVF and embryo transfer with frozen-thawed semen. With fresh semen, spermatozoa from one cloned bull had lower motility than its donor. Cloned bulls had higher velocity parameters in fresh semen, but those effects were not obvious in frozen-thawed or frozen-thawed semen selected with a Percoll gradient. Semen collected from cloned bulls had significantly higher IVF rates compared to donors; however, embryo development per cleaved embryo or quality of blastocysts did not differ between donors and cloned bulls. Pregnancy and live offspring rates from one donor and its cloned bull did not differ between fresh (40%, 16/40 versus 46%, 17/37) and vitrified/thawed (13%, 2/16 versus 25%, 4/16) embryo transfer following IVF. A total of 26 calves were obtained from genotypically identical donor and cloned bulls with no signs of phenotypical abnormalities. These preliminary results suggested that the physiology of surviving postpubertal cloned bulls and quality of collected semen had equivalent reproductive potential to their original cell donor, with no evidence of any deleterious effects in their progeny.     

Phenotypic relationships between hair whorl characteristics and spermatozoal attributes in Holstein bulls

The objective of this study was to characterize facial whorl characteristics in Holstein stud bulls (n = 485) and to investigate their association with measures of semen quantity and quality. Bulls had 1 (n = 454), 2 (n = 21) or no facial hair whorls (n = 5). Some 39% of bulls had whorls in the middle of their forehead, whereas 43% had whorls very low down on their forehead. Spiral whorls were present on 77% of bulls while 22% had elongated whorls. Abnormally shaped whorls were found more often on bulls with low whorls (27% versus 17%, P = 0.01) than on bulls with high or middle whorls. Semen production data were obtained from the AI center database to provide a representative sample of each sires most recent semen production characteristics. The number of semen collections per bull averaged 38 with a range of 1 (n = 9) to 272 (n = 1). The average number of days between collections was 5 but varied from 1 to 425 days. Average age of bulls at first recorded collection was 679 days and ranged from 307 to 4555 days. There were no significant differences between black and white bulls in semen quality or quantity. Semen quality over the summer months was reduced in comparison to the winter months. Age of the bull at time of collection had no effect on any of the semen quality traits. Older bulls had larger scrotal circumference and produced more semen (P > 0.01). Bulls with whorls located in the center of the forehead were not significantly different in semen quality or quantity for the 10 traits considered when compared to bulls with whorls located outside the center. In contrast to previous research findings with Angus cattle, semen attributes were not significantly different between bulls characterized with round (or spiral) epicenters and those with abnormally shaped whorls. The difference between the two studies may be due to the fact that Angus bulls have a higher percentage of abnormal elongated whorls compared to Holsteins.     

Novel Chlamydiales strains isolated from a water treatment plant

Chlamydiae are obligate intracellular bacteria infecting free-living amoebae, vertebrates and some invertebrates. Novel members are regularly discovered, and there is accumulating evidence supporting a very important diversity of chlamydiae in the environment. In this study, we investigated the presence of chlamydiae in a drinking water treatment plant. Samples were used to inoculate Acanthamoeba monolayers ( Acanthamoeba co-culture), and to recover autochthonous amoebae onto non-nutritive agar. Chlamydiae were searched for by a pan-chlamydia 16S rRNA gene PCR from both Acanthamoeba co-cultures and autochthonous amoebae, and phylotypes determined by 16S rRNA gene sequencing. Autochthonous amoebae also were identified by 18S rRNA gene amplification and sequencing. From a total of 79 samples, we recovered eight chlamydial strains by Acanthamoeba co-culture, but only one of 28 amoebae harboured a chlamydia. Sequencing results and phylogenetic analysis showed our strains belonging to four distinct chlamydial lineages. Four strains, including the strain recovered within its natural host, belonged to the Parachlamydiaceae ; two closely related strains belonged to the Criblamydiaceae ; two distinct strains clustered with Rhabdochlamydia spp.; one strain clustered only with uncultured environmental clones. Our results confirmed the usefulness of amoeba co-culture to recover novel chlamydial strains from complex samples and demonstrated the huge diversity of chlamydiae in the environment, by identifying several new species including one representing the first strain of a new family.     

The use of in vitro fertilization techniques to investigate the fertilizing ability of bovine sperm with proximal cytoplasmic droplets

The objective of this experiment was to determine the effect of proximal droplets on sperm-oocyte binding, zona penetration, fertilization, and the developmental competence of oocytes fertilized by sperm with proximal droplets (PD) in an in vitro fertilization (IVF) and culture system. Frozen semen from three bulls (PD1, PD2 and PD3) with varying proportions of normal appearing sperm with proximal droplets and semen from a normal control bull (C) were used in this experiment. The mean number of sperm bound to the zona pellucida (26.8 +/- 2.0, n=100; 15.2 +/- 1.1, n=100; 16.2 +/- 1.0, n=100) for bulls PD1, PD2, and PD3, respectively, were significantly lower (P<0.05) than that of the control bull C (47.4+/- 1.9; n=114). No spermatozoa with PD were found bound to the zona pellucida and this finding was consistent among the three bulls. The percentage penetration of zonae for the bulls PD1, PD2 and PD3 (74%, 74/100; 71%, 71/100 and 69%, 69/100, respectively) were not different than that of bull C (72%, 179/245). Similarly, the mean number of sperm penetrating the zona pellucida (1.43+/- 1.2, 1.24 +/- 1.1 and 1.20 +/- 1.1, for bulls PD1, PD2 and PD3, respectively) were not different than that of bull C (1.45 +/- 1.1). However, fertilization rates (8.8%, 8/90; 16.8%, 16/95; and 10.6%, 11/103, for bulls PD1, PD2 and PD3, respectively) were lower (P<0.001) than that of bull C (68.7%; 77/112). Similarly, cleavage rates (5%, 10/200; 8%, 8/100 and 14%, 15/111) for the bulls PD1, PD2 and PD3, respectively, were lower than that of the control bull, C (60.7%; 79/130). Cleaved zygotes resulting from the fertilization of oocytes by apparently normal sperm from bulls with PD did not develop beyond cleavage, whereas, 43.8% (57/130) morulae and 20% (26/130) blastocysts were produced by oocytes fertilized by sperm from bull C. In summary, normal appearing sperm with PD did not bind to the zona pellucida. Apparently normal sperm with out proximal droplets co-existing in the semen along with sperm containing PD were also functionally deficient, resulting in reduced zonae binding and zygotes resulting from insemination with semen with a high percentage of PD did not develop beyond cleavage.     

Amplification and Characterization of Bull Semen Infected Naturally with Foot-and-mouth Disease Virus Type Asial by RT-PCR

To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.     

Breed differences in competitive indices of Holstein and Jersey bulls and their association with sperm DNA fragmentation index and plasma membrane integrity

The objective was to evaluate the relationship of a competitive index (CI) determined by heterospermic performance and post-thaw semen quality of the same stored ejaculates. Semen from multiple ejaculates collected in succession from each bull (four Holstein and four Jersey) was pooled. Heterospermic doses (20x10(6)/straw) were made to obtain all possible Holstein-Jersey combinations (16 two-bull combinations) and contained 20x10(6) sperm/mL/bull. Cows at two University dairy farms were inseminated on observed or synchronized estrus. The sire of calves (N=460) were determined and a CI was determined for each bull (based on the number of calves sired). Prior to preparation of the heterospermic doses, a sub-sample of semen from each bull was taken, processed, frozen, and stored concurrently with heterospermic samples. Post-thaw semen samples (homospermic) from each bull were assessed for: sperm morphology, acrosome integrity, sperm motility parameters assessed by computer assisted sperm analysis (CASA), flow cytometry analysis of DNA Fragmentation Index (DFI), and Plasma Membrane Integrity (PMI). Heterospermic performance of Holstein bulls was superior to that of Jersey bulls. The DFI was negatively correlated to CI (r=-0.87; P<0.001), whereas the PMI (r=0.87; P<0.001) and total progressive motility (r=0.74; P<0.05) assessed by CASA were positively correlated to CI. In multivariate regression models, the DFI and PMI accounted for 87% variance in competitive index. In conclusion, bulls with less DFI and higher PMI had higher probabilities of siring calves.     

Distribution of Corynebacterium renale among healthy bulls with special reference to inhabitation of type III in the prepuce.

Distribution of Corynebacterium renale among apparently healthy bulls reared in Hokkaido was investigated. The organism was detected from 46 (39.3%) of 117 specimens of preputial cavity washing and from 60 (51.7%) of 116 specimens of semen. The isolates studied in this survey belonged to type III, except a few which belonged to type II. No type I strain was isolated from any bull. C. renale type III was isolated from the prepuce in six of seven bulls slaughtered and from urethra in three, but not at all from any other organ. In the seven bulls, no macroscopic changes were seen, but a slight infiltration of lymphocytes and formation of lymph nodules were noticed in the prepuce. No other microscopical changes could be demonstrated in any other organ. No serum antibody response was detected. To ascertain the virulence of C. renale isolated from the bulls, a strain of type II was inoculated into the urinary bladder of a healthy cow. The cow exhibited fever and hematuria on and after the 10th day. Typical cystitis was proved when the cow was necropsied on the 14th day after inoculation. From these result it is conceived that C. renale type II organisms inhabit the prepuce of apparently healthy bulls at a high rate, without inducing any disturbance.     

Effect of age and environmental factors on semen quality, glutathione peroxidase activity and oxidative parameters in simmental bulls

Taking into account that semen quality depends on animal age and climate conditions and that oxidative stress has been reported to be a common cause of infertility, the objective of this study was to monitor indicators of oxidative stress and antioxidant protection during four seasonal periods in service bulls of various age to get better insight into the significance of these factors upon evaluating service bull semen. The research was conducted over a year on 19 Simmental service bulls. Animals were divided into two groups according to age; Group I consisted of younger bulls aged two to four yrs (n = 9), and Group II was comprised of older bulls aged five to ten yrs (n = 10). Semen samples were obtained once in the middle of every seasonal period and blood samples for biochemical analysis were collected by jugular venipuncture immediately after ejaculate collection. The activity of total glutathione peroxidase (T-GSH-Px), selenium-dependent glutathione peroxidase (Se-GSH-Px) and selenium-independent glutathione peroxidase (non-Se-GSH-Px), together with the intensity of lipid peroxidation (thiobarbituric acid reactive substances; TBARS) and oxidative protein damage (protein carbonyl content (PCC)) were measured in seminal plasma. In samples of spermatozoa and blood serum, the activity of Se-GSH-Px and TBARS and PCC concentrations were determined. Older service bulls had significantly higher ejaculate volume in summer in comparison with younger bulls, whereas the number of spermatozoa and progressive motility percentage did not significantly vary with age. Younger animals had lower progressive motility percentage during summer than in spring, with more intensive oxidative processes observed in seminal plasma (TBARS) and spermatozoa (TBARS and PCC). Based on the results presented here, it can be concluded that younger bulls are more sensitive to elevated ambient temperatures during the summer, when intensified prooxidative processes in semen plasma and spermatozoa eventually led to decreased sperm progressive motility with consequential semen quality deterioration.     

Occasional detection of Neospora caninum DNA in frozen extended semen from naturally infected bulls

Recently, the presence of Neospora caninum DNA in semen from naturally infected bulls was reported. In the present work, the presence and quantification of N. caninum by PCR techniques in frozen extended semen straws from naturally infected bulls was investigated. A total of 20 seropositive and five seronegative bulls raised for reproductive purposes in an AI centre were used. Ten extended semen straws from each bull obtained at different time-points during the previous 2 years were selected for Neospora testing. Eight of the seropositive bulls (40%) studied showed at least one positive straw to N. caninum DNA and 14 of their 180 semen straws examined (7.8%) were found to be positive. In all positive samples, N. caninum DNA was consistently detected in the cell fraction and not in the seminal plasma. However, the parasite number in each positive straw was under the detection level of real-time PCR. In parallel, 10 semen straws from each of the five seronegative bulls were also analyzed by the nested-PCR and no N. caninum DNA products were obtained. In order to check the consistent presence of N. caninum in a positive semen batch, three additional semen straws from the same batch of each positive straw from three seropositive bulls were analyzed but N. caninum DNA was only detected in one straw from one bull. In conclusion, we report the sporadic detection of N. caninum DNA in semen straws of naturally infected bulls but the low frequency of contaminated semen straws and the low parasite load observed indicate a minor chance of bovine neosporosis transmission by AI.     

Amplification and characterization of bull semen infected naturally with foot-and-mouth disease virus type Asia1 by RT-PCR

To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group. Foundation items: State Science and Technology Support Program (2006DAD06A03) and Hi-tech Research and Development Program of China 863 (2006AA10A204).     

Effects of the anabolic steroid, boldenone undecylenate, on certain reproductive parameters in bulls

A total of 17 bulls was used to study the effects of boldenone undecylenate on growth and semen characteristics in beef bulls. In trial 1 nine mature mixed-breed beef bulls with satisfactory semen quality were divided into two groups. Group I (n = 5) received boldenone undecylenate (1.1 mg/kg) at 21 d intervals for a total of seven treatments (147 d). Group II (n = 4) served as untreated controls. Semen was collected from each group by electroejaculation on each treatment day and evaluated according to the standards of the Society for Theriogenology. Although neither the percentage of spermatozoa with primary or secondary morphological abnormalities was different, the ejaculates of Group I bulls contained a higher percentage of abnormal spermatozoa than those in Group II. In trial 2, eight mixed-breed bull calves, average weight 140.4 kg, were maintained under drylot conditions in a single paddock. The bulls were divided into two equal groups. Group I (n = 4) received boldenone undecylenate as in Trial 1. Group II (n = 4) served as untreated controls. The bulls were weighed and the scrotal circumference (SC) was measured every 21 d until it reached 30 cm, at which time semen was collected and evaluated as in Trial 1. Group I bulls had a higher percentage of spermatozoa with primary morphological abnormalities than bulls in Group II. Group I bulls had a higher average daily gain (ADG) than Group II bulls and required 21 d longer for the SC to reach 30 cm. Semen quality for all bulls was satisfactory at each sampling day.     

Inoculation of bulls with bovine virus diarrhea virus: Excretion of virus in semen and effects on semen quality

Bovine virus diarrhea (BVD) virus was inoculated into 9 bulls. Semen samples were collected to determine if BVD virus could be isolated from semen and to determine if the virus affected semen quality. The reproductive tracts of 6 bulls were recovered following slaughter and subjected to virus isolation procedures and histologic examination.BVD virus was detected in 4 of 98 semen samples following inoculation. The virus was also recovered from the testicle of one bull following slaughter. Inoculation of BVD virus into bulls did not affect semen quality and did not cause gross or microscopic lesions in the reproductive tract. This experiment indicates that following inoculation of BVD virus into bulls, the virus may be shed in the semen. However, the probability of this occurring is low.