Prevalence of Tritrichomonas foetus in several subpopulations of Alabama beef bulls.

The objective of this study was to estimate the prevalence of Tritrichomonas foetus infections in Alabama (USA) beef bulls through prospective and retrospective surveys. The prospective survey included 240 Alabama beef bulls that were sampled between January 2005 and March 2006. Preputial smegma was collected from the 240 bulls with a dry pipette and cultured in an InPouch TF T. foetus culture pouch (BioMed Diagnostics; White City, OR, USA). The samples were evaluated microscopically once a day for 6 days for growth resembling T. foetus. To avoid false-positives due to fecal trichomonads, all suspect cultures were sent to both the Alabama Department of Agriculture Veterinary Diagnostic Laboratory in Auburn, AL, USA and the Auburn University College of Veterinary Medicine Parasitology Laboratory (Auburn, AL, USA) for polymerase chain reaction (PCR) confirmatory assays. Of the 240 bulls cultured in the prospective survey, 3 (1.25%) cultures were considered suspect on microscopic evaluation. However, PCR-based assays were negative for T. foetus, suggesting that the samples most likely contained fecal trichomonads. The retrospective analysis included 374 T. foetus cultures performed at the Alabama Department of Agriculture Veterinary Diagnostic Laboratory between October 2002 and March 2005. Of the 374 bulls included in the retrospective analysis, only 1 (0.27%) was confirmed positive by a PCR-based assay.

Review: Abnormalities of the bull - occurrence, diagnosis and treatment of abnormalities of the bull, including structural soundness.

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.

Initial steps in defining the environment of the prepuce of the bull by measuring pH and temperature.

OBJECTIVE: To determine the baseline pH and temperature of the preputial cavity of bulls. METHODS: We enrolled 55 bulls ranging in age from 15 to 84 months. The preputial temperature and pH were measured by insertion of temperature and pH probes, respectively, into the preputial orifice prior to routine breeding soundness examinations. Information was obtained from owners regarding the diet of each bull and categorised as one of three categories: forage only, grain supplemented or silage supplemented. RESULTS: The average temperature of the prepuce was 37.81°C ± 1.76 and the median pH of the prepuce was 8.45 (6.35-9.46). Preputial temperatures of the bull weakly correlated with ambient temperatures (rs = -0.29, P = 0.028). The preputial pH of silage-fed bulls was significantly lower than that of bulls fed forage only (P = 0.025) or grain-supplemented diets (P = 0.002). The median preputial pH of bulls fed a silage-based diet was 7.6 (6.3-8.9) compared with a median pH 8.7 (7.8-9.1) for bulls fed forage-based diets or a median of 8.5 (7.7-9.4) for those given grain-supplemented diets. CONCLUSION: Diet and ambient temperature can, respectively, affect pH and the temperature in the prepuce. Further studies to describe and understand the microbiota of the prepuce and penis may assist in developing treatments for diseases of the genital tract in bulls.

Characterization of seminal plasma proteins and sperm proteins in ejaculates from normospermic bulls and bulls with thermally-induced testicular degeneration.

Semen was collected from 5 mature beef bulls by electroejaculation before, during, and after 20 days of scrotal insulation. Thermally-induced testicular degeneration was irreversible in 3 of the bulls. Analysis of sperm and seminal plasma polypeptides revealed that 15 to 30 sperm polypeptides and 25 to 30 seminal plasma polypeptides were indistinguishable between bulls prior to the insulation treatment. Changes in the sperm polypeptides pattern appeared as early as 2 days after the insulation treatment and persisted for at least 11 months in 2 of the bulls. In the spermatozoa, there was a detectable loss of 31, 34, 49 and 58 kDa polypeptides and an appearance of 6 to 8 new major polypeptides, ranging from 32 to 83 kDa. The 83 kDa polypeptide was most prominent in the 2 bulls that regained normal sperm motility and morphology following the insulation period. The post-insulation appearance of a seminal plasma polypeptide (circa 60 kDa) was also identified in these 2 bulls. Seminal plasma polypeptides remained qualitatively unaltered by the insulation treatment in the 3 bulls with irreversible testicular degeneration.

Hematologic and semen quality changes in bulls with experimental eperythrozoon infection.

Eperythrozoon organisms were isolated from the blood of a young beef bull with scrotal and hindlimb edema. Young beef bulls, managed under conditions closely mimicking those used in organized bull testing programs, were experimentally administered eperythrozoon organisms. Scant to few organisms were identified on blood smears from bulls (5 of 6) for 2 to 4 days starting 12 days after administration. After a second challenge with intravenously administered viable eperythrozoon organisms, the bulls demonstrated immunity by either failing to become parasitemic (4 of 6) or rapidly clearing the organisms from the blood (2 of 6). No bull became anemic, icteric, or hypoglycemic. Increased serum lactate and decreased blood bicarbonate concentrations probably reflected increased glycolytic activity of infected erythrocytes. A cause for azotemia observed late in the study was not determined. The bulls did not develop scrotal or hindlimb edema. Scrotal circumference and texture remained constant throughout the study. Semen quality was minimally altered while the bulls had organisms identified on blood smears. Harsh lung sounds were asculted in bulls during and immediately after organisms were present in the blood. Although the bulls reliably had organisms in the blood, none showed anemia, scrotal or hindlimb edema, or decreased semen quality; therefore, an additional factor or factors, or greater parasite load may be required for the expression of disease.

Eperythrozoon infection in young bulls with scrotal and hindlimb edema, a herd outbreak.

Young (9 to 10 mo) Aberdeen-Angus bulls (n = 5) in excellent body condition and pastured with approximately 35 other bulls of similar age on a farm in southeastern Alabama had fever, harsh pulmonary sounds, increased respiratory rate and variable amounts of hindlimb and scrotal edema. Bulls had mild microcytic, normochromic anemia. Numerous eperythrozoon organisms were identified on blood smears. Indirect hemagglutination inhibition test results for Eperythrozoon suis antibodies were negative either because E. suis antigens do not cross react with cattle eperythrozoon organism antibodies or blood was collected before there was sufficient time for seroconversion. Bulls had swelling of the scrotal wall, soft testes, and poor semen quality, characterized by low progressive motility and a high percentage of spermatozoa with primary and secondary abnormalities. Some of these abnormalities may be attributed to the age of the bulls. However, loss of scrotal thermoregulation was a major cause of testicular degeneration leading to poor semen quality. Other bulls in the herd had fever, increased respiratory rate, and swollen hindlimbs or scrotum. Subsequent to administration of oxytetracycline, parasitemia resolved rapidly, and clinical signs gradually abated. Four of five bulls successfully passed breeding soundness examinations six months after initial clinical illness. The remaining bull failed twice due to the presence of testicular inflammation.

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.

The effects of ovarian hormones and ACTH on uterine defense to Corynebacterium pyogenes in cows.

Ovariectomized cows were treated with either estrogen, progesterone or adrenogorticotropic hormone (ACTH). Each cow's uterus was inoculated with 1x10(6)Corynebacterium pyogenes organisms. The cows were evaluated by rectal examination for signs of uterine infection. Peripheral leukocytic phagocytosis was determined by chemiluminescence at the beginning (Day 4) and end (Day 21) of the experiment. Uterine cultures were obtained at the end. None of the estrogen-treated cows developed signs of uterine infection, but all the other cows did develop uterine disease. All the infected cows showed clinical improvement at the experimental end (Day 21). There were no differences between groups for leukocytic phagocytosis on Day 4, but on Day 21, values for progesterone-treated and control cows were similar but greater than those for estrogen or ACTH-treated cows, which were similar. Leukocytic phagocytosis values for all cows were lower on Day 21 than on Day 4. Most of the estrogen- and ACTH-treated cows had negative intrauterine cultures on Day 21 while most of controls and progesterone-treated cows had positive cultures.

Adherence of Brucella ovis to preimplantation ovine ova.

Seventy-three zona pellucida-intact ova were collected surgically from 15 superovulated, Brucella -free mixed-breed ewes. Twenty-one groups containing one to five ova were incubated in medium containing Brucella ovis . Subsequently, seven and five groups were incubated for 24 and 4 h, respectively, at 37 degrees C in medium containing penicillin and streptomycin, while nine groups were not treated with antibiotics. All groups of ova were washed 10 times, and ova and sequential washes were cultured for the presence of B. ovis . Brucella were isolated from seven of the nine groups of nontreated ova and from the 10th wash for six of these groups. While Brucella were detected in fewer washes after antibiotic treatment, the organism was still isolated from 11 of the 12 treated groups. Results indicate that standard washing techniques are not reliable for removing B. ovis from exposed, zona pellucida-intact, ovine ova.

Seroconversion of recipient ewes after transfer of ova exposed to Brucella ovis in vitro.

Zona pellucida-intact ova collected from ewes seronegative to Brucella ovis were exposed in vitro to B ovis and washed 10 times in medium that contained no antibiotics. After exposure and washing, nontransferable ova were cultured for isolation of Brucella , and the viable ova were transferred into seven B ovis seronegative ewes. No pregnancies resulted, thus recipient ewes were bred during the next breeding season, and blood samples were collected for bacteriological and serological examination until one month after lambing. Brucella ovis was isolated from all of the nontransferable ova, indicating that the transferred ova had viable organisms adhered to them. Although no recipient was found to be pregnant at Day 45, all seven ewes responded to the transferred ova by producing anti-Brucella antibodies. With the exception of a ewe that was euthanized early in the project due to a traumatic injury, all recipients lambed normally during the following breeding season. Brucella was not found in any sample collected from ewes or lambs. However, ELISA titers for B ovis remained in the suspicious range and a ewe was positive on the CF test within 2 wk of lambing.

In vitro exposure of ovine ova to (Brucella abortus ).

Fifty-three zona pellucida-intact ova were collected surgically from superovulated, Brucella -free mixed-breed ewes. Groups containing two to seven ova were incubated in medium containing Brucella abortus . All groups of ova were then washed 10 times, and ova and sequential washes were cultured for the isolation of B. abortus . Brucella were not found beyond the fifth wash for any group of ova, but were isolated from one of 12 groups of ova. Results indicate that mechanical washing in the absence of antibiotics is advantageous, but alone, is not totally reliable for removing B. abortus from exposed, zona pelucida-intact ovine ova.

A caudal flank approach for the collection of oviductal-stage bovine embryos.

Oviductal-stage embryos were surgically collected from 27 superovulated adult cows of various breeds, ages, and parity. A total of 88 surgeries was performed via a caudal flank grid approach, with the animals in lateral recumbency and the reproductive tract irrigated with sterile glycerol solution prior to surgical closure. Eight cows were operated on twice and five cows were operated on three or more times. The maximum number of surgeries for a single cow was five. Successful ova collection was accomplished in each surgical attempt, and all cows submitted to this procedure subsequently became pregnant following return to the breeding herd. This technique provided greater exposure of the ovary, uterine tube, and uterine horn, with less adhesion formation than traditional ventral midline techniques.

Effects of embryo-freezing and thawing techniques on the survivability of Brucella abortus.

A suspension of a pathogenic strain (2308) of Brucella abortus was aliquoted, centrifuged, resuspended in 6 treatment media and quantitated. Ten 1-ml samples of each treatment were subjected to a standard embryo-freezing technique. The treatments were selected to examine the effects of 1) freezing and thawing, 2) cryoprotectants and 3) antibiotics on the survivability of Brucella suspended in embryo-support media. Five samples of each treatment were thawed and quantitated after a 2-wk storage period and five samples were thawed and quantitated after a 6-mo storage period. Means and percent reductions were determined for each treatment. There was no statistical difference between means at 2 wk and 6 mo within any treatment. Freezing and thawing caused a 64% reduction in the number of viable Brucella . The addition of antibiotics caused a 99.9% reduction in viability of the organism. Glycerol protected the organism during freezing and thawing in the absence of antibiotics but did not interfere with the high percent reduction seen when antibiotics were present. Dimethylsulfoxide (DMSO), however, not only protected the organism during freezing and thawing but also appeared to negate the effects of the antibiotics.

Effect of transportation stress on ovarian function in superovulated Hereford heifers.

A study was conducted to determine the effect of transportation stress on ovarian function in superovulated heifers. Thirty cyclic Hereford heifers of similar age and weight and in good body condition were randomly assigned to control and stress-treated groups. All animals received two daily injections of 5 mg follicle stimulating hormone (FSH) for 4 d beginning on Day 10 to 12 of the estrous cycle. A blood sample was collected at each FSH injection. On the fourth day of injections, heifers were given 25 mg prostaglandin F(2)alpha (PGF(2)alpha) in the morning and a second injection of 15 mg PGF(2)alpha in the afternoon. During superovulation, the stressed heifers were transported to a different location every 12 h whereas control animals remained at the pretrial site. Following 4 d of intermittent transporting and FSH treatment, stress-treated heifers were recombined with control animals. Ovaries were examined 8 d following the onset of standing estrus to determine length, width, thickness, and number of corpora lutea (CL). Peripheral plasma levels of cortisol were higher in the stressed group (P< 0.1). Least squares means for numbers of CL were 20.4 +/- 2.1 and 15.4 +/- 1.7 for control and treated heifers, respectively (P< 0.1). There were no treatment differences (P> 0.1) between length, width, or thickness of ovaries when the number of CL was held constant. These data suggest that stress of the type, intensity, and duration imposed in this study increased plasma levels of cortisol and reduced ovulation rate as determined by CL formation in superovulated heifers.

Embryo transfer from goats seropositive for caprine arthritis-encephalitis virus.

Twelve attempts were made to isolate caprine arthritis-encephalitis virus (CAEV) from the uterine flushings of serologically positive superovulated does mated to serologically positive bucks. Embryos were transferred to eight serologically negative estrus-synchronized recipient does and the recipients were monitored serologically following embryo transfer. Virus isolation was attempted from colostrum and placental tissues from does that kidded following embryo transfers and the surviving kid was monitored serologically until four months of age. The CAEV was not isolated from any of the uterine flushings, colostrum or placental tissues. All recipients and the kid remained seronegative throughout the trial.

Testicular shape and its relationship to sperm production in mature Holstein bulls.

This study was conducted to determine the relationship between testicular shape, scrotal circumference (SC) and sperm production. Twenty-seven mature Holstein bulls were evaluated subjectively and objectively for testicular shape as indicated by testicular length and width, then placed in 1 of 3 groups. Group 1 contained 17 bulls with a normal ovoid testicular shape and a length to width ratio of 1.61:1 +/- 0.01 (SEM). Group 2 was composed of 4 bulls with a long, slender testicular shape and a length to width ratio of 1.95:1 +/- 0.06 (SEM). Group 3 was comprised of 6 bulls with spheroid-shaped testicles and a length to width ratio of 1.3:1 +/- 0.03 (SEM). All the groups were statistically different for length to width ratios (P < 0.05). Length measurements from cranial to caudal pole of the testis proper were also different between groups (P < 0.05). Width or testicular diameter was different between Group 2 and Group 3 at P < 0.05; however, there was no difference between Group 1 and Group 2 or between Group 1 and Group 3. Predicted volumes and weights of testicles were not significantly different between groups. Scrotal circumference measurements were significantly different between groups (P < 0.05). Group 1 had an average SC of 43.07 +/- 0.36 cm (SEM), Group 2 of 39.33 +/- 1.18 cm (SEM) and Group 3 of 46.22 +/- 0.69 cm (SEM). Sperm production for a twice daily, 2-day-per-week collection schedule revealed a statistically significant difference for sperm output. A total of 2742 ejaculates was evaluated. A total of 1818 ejaculates was evaluated in Group 1, 440 ejaculates in Group 2 and 484 ejaculates in Group 3. The mean spermatozoal harvest per day for Group 1 bulls was 13.62 +/- 0.09 x 10(9) (SEM). Group 2 bulls with the longer-shaped testicles produced 14.82 +/- 0.18 x 10(9) (SEM) spermatozoa per day, and Group 3 bulls, with the more rounded testicle shape and the significantly larger SC produced 11.72 +/- 0.64 x 10(9)(SEM) sperm cells per day. All 3 groups were statistically different at the P = 0.05 level. The results suggest that prediction of sperm production may be dependent on factors other than SC, testicular volume, or weight. Testicular shape may influence sperm output in mature Holstein bulls.

Blood flow to the corpus cavernosum penis in the bull and goat buck during penile quiescence.

Bulls and bucks were used to study the blood flow into and out of the corpus cavernosum penis (CCP) during the non-erect state of the penis. When contrast medium was injected into the dorsal artery of the penis in a bull cadaver and into surgically implanted catheters of anesthetized bucks, it flowed into the cavernous spaces of the CCP via the penetrating arteries. When contrast medium was injected directly into the CCP of the bull and buck there was no evidence of vascular exits along the body of the penis. The only venous outlets from the CCP were in the crus penis area. In the non-erect penis the vascular pressure within the CCP was 17.8 mm Hg higher in the bull and 8.3 mm Hg higher in the buck at the glans area than the crus area. During the non-erect state of the penis most of the blood flow to the CCP is from the dorsal artery of the penis via the penetrating arteries. The results showed that the pressure gradient provides the means for flow of blood from the glans area toward the base or crus penis area. This flow prevents platelet aggregation and clot formation and provides nutrition to local tissue during the long periods of non-erection.

Caliper and ultrasonographic measurements of bovine testicles and a mathematical formula for determining testicular volume and weight in vivo.

This study quantified the relationship between calibrated caliper and ultrasonographic derived measurements of bovine testicles in vivo with actual testicular length, width, volume and weight. The prolate spheroid formula was tested to accurately predict testicular volume and a modification to predict weight. Ten bulls were employed to derive caliper and ultrasound testicle (n = 20) length and width measurements in vivo. Caliper length measurements were more reliable than ultrasound derived lengths, with correlations of r2 = 0.8023; P < 0.05 and r2 = 0.5111; P < 0.05, respectively. Width for both the calipers and ultrasound measurements when compared to actual width measurements were r2 = 0.7313; P < 0.05 and r2 = 0.8310; P < 0.05, respectively. The prolate spheroid formula is reliable in determining testicle (n = 116) volume (r2 = 0.8928; P < 0.05). Testicular volume and weight are highly correlated (r2 = 0.9776; P < 0.05); therefore, a modification of the prolate spheroid formula was used to predict weight (r2 = 0.9084; P < 0.05) against the actual weight. Caliper-derived length and width measurements used in the prediction of volume and weight had correlation coefficients against actual volume and weight of r2 = 0.5497; P < 0.05 and r2 = 0.6340; P < 0.05, respectively. Ultrasound in vivo measurements for prediction of testicular volume and testicular weight had a correlation of r2 = 0.3276; P < 0.05 and r2 = 0.6249; P < 0.05, respectively. A testicular (n = 116) length to width ratio of 1.8:1 (SEM = 0.01) was determined for both slaughterhouse and castrated animals. Caliper measurements are reliable, inexpensive and much simpler to obtain than ultrasound determinations for in vivo testicle length, width, volume and weight. The two-dimensional measurement of length and width would be a more accurate predictor of testicle volume and weight than the one-dimensional measurement of scrotal circumference (SC), especially in bulls with variation in testicular shape.

Ovine uterine morphogenesis: effects of age and progestin administration and withdrawal on neonatal endometrial development and DNA synthesis.

To determine effects of age and administration and withdrawal of a synthetic progestin (P) on endometrial development and DNA synthesis, ewe lambs were ovariectomized on d 0 (birth) and assigned to one of four groups (n = three/group) that provided (by means of hemihysterectomy) the following uterine tissue types: 1) d 0 control, 2) d 13 control, 3) d 26 control, 4) d 13 after 13 d exposure to P (13P) and 5) d 26 after P exposure from d 0 to 13 (26P). Uterine tissues were processed for histology or explanted with [methyl-3 H] thymidine for autoradiography. Labeling index (LI) was determined for stroma and epithelium in caruncular and intercaruncular endometrial areas and for lumenal and glandular epithelium in uteri with glands. Endometrial glands were absent on d 0, evident at d 13 and well developed by d 26. Day 0 LI was greater (P less than .05) in caruncular than in intercaruncular areas, and greater in stromal than in epithelial tissues. Relationships were reversed in d 13 endometrium (day X endometrial area, P less than .07). Caruncular stromal LI was greater on d 0 than later (P less than .02), whereas intercaruncular epithelial LI was greater after d 0 (P less than .05), but decreased from d 13 to 26 (P less than .05). Glandular epithelial LI was higher on d 13 than on d 26 (P less than .03). Administration of P inhibited endometrial gland development and suppressed d 13P intercaruncular LI (P less than .05). Withdrawal of P was followed by endometrial gland development and increased (P less than .01) intercaruncular epithelial LI in d 26P uteri. Ovary-independent initiation of endometrial gland development involves age- and region-specific alterations in DNA synthesis and could involve negative control.

Comparison of urea treatment with established methods of sorghum grain preservation and processing on site and extent of starch digestion by cattle.

To determine the effect of dry (D); reconstituted and ensiled (R); reconstituted and acid-treated (A); and urea-treated, high-moisture (U) sorghum grain on starch digestibility, four Angus x Hereford steers (means BW = 350 kg) with duodenal and ileal cannulas were used in a 4 x 4 Latin square design. Diets consisting of 69% ground sorghum grain were fed every 2 h in equal portions (8.2 kg/d). Diets averaged 46.5% starch and 12% CP, except for U, which averaged 14% CP due to urea treatment. Ytterbium attached to sorghum was used as a particulate marker. Duodenal, ileal, and fecal samples were taken 1 h postfeeding after a 14-d adaption to diets. Whole samples were analyzed. Preduodenal starch digestion (%) was 89, 83, 76, and 70, and starch digestion over the total tract was 99, 97, 95 and 91 for R, U, A, and D, respectively. Starch digestion proximal to each site (duodenum and ileum) was enhanced (P less than .05) by R and U compared with D. Within the small intestine, there was a linear relationship (P less than .003) between starch digestion and daily starch supply. However, digestibility of starch in the small intestine (mean = 45%) was not different among diets. Apparent digestibility of starch in the large intestine was not significantly different from digestibility in the small intestine. Urea-treated sorghum grain was equivalent to reconstituted, ensiled sorghum in digestion characteristics and was superior to dry sorghum.