Aerobic bacterial flora of the vagina and prepuce of California sea lions (Zalophus californianus) and investigation of associations with urogenital carcinoma

To investigate the association between genital bacterial infection and urogenital carcinoma in California sea lions (Zalophus californianus), vaginal and preputial swabs for bacterial isolation were taken from 148 free-ranging and 51 stranded California sea lions including 16 animals with urogenital carcinoma. Cytological examination of vaginal or preputial smears showed a majority (65.5%, 57/87) of animals examined had mild or no inflammation. Aerobic bacteria were isolated from 116 (78.4%) wild sea lions and 100% of stranded animals. A total of 403 isolates were identified representing 51 unique bacterial species. The median number of isolates per animal increased with age in the wild group, but there was no difference in the number of isolates per animal between wild and stranded adults. The most common bacteria isolated from the wild sea lions were Psychrobacter phenylpyruvicus (39 isolates), non-hemolytic Streptococcus (35 isolates), Corynebacterium spp. (30 isolates), and Escherichia coli (20 isolates). More bacterial species were isolated from stranded animals than wild animals (33 versus 26) and there was significantly less growth of P. phenylpyruvicus, Corynebacterium spp., and Moraxella-like spp. in the stranded animals. Beta-hemolytic Streptococcus was the only bacterium significantly associated with urogenital carcinomas in California sea lions, but only in females.     

The effect of pseudorabies (Aujeszky''s) virus infection on young mature boars and boar fertility.

This study was designed to determine the effects of experimental inoculation with pseudorabies virus on the reproductive tracts of young adult boars. Pseudorabies virus was inoculated intranasally into 12 boars and intrapreputially into four boars. All animals seroconverted after nasal or preputial inoculation. Semen abnormalities were observed 21 days postinoculation with partial recovery by 50 days postinoculation. Virus was isolated from the preputial sheath of two intrapreputially inoculated boars 12 days postinoculation. It was concluded that pseudorabies virus infection can be established via preputial inoculation and that decreased spermatogenesis and infertility can result.     

Collection of preputial material by scraping and aspiration for the diagnosis of Tritrichomonas foetus in bulls

Two trials were carried out to assess the diagnostic sensitivity and practicability of preputial scraping as a method of collecting preputial material from bulls infected with Tritrichomonas foetus. In the 1st trial, preputial material was collected by simultaneous scraping and aspiration from 3 infected and 1 uninfected bull 10 times over a 5-week period. In the 2nd trial, samples from 5 infected bulls were collected by both sheath washing and scraping on 6 occasions, while 8 uninfected animals were sampled 3 times. Samples were cultured using a modified Trichomonas culture medium (Oxoid). In the first trial, 29 of 30 samples from infected bulls were found to be positive. In the second trial, 83 % of samples collected by both methods tested positive. In neither trial were any samples from the control bulls found to be positive. Scraping was found to be quick and safe, and offered advantages over preputial washing in that urine contamination was easily avoided, samples were smaller and more concentrated and contamination was reduced. It may, however, be subject to greater operator variability than sheath washing. It is concluded that preputial scraping is as effective as washing and represents a suitable alternative for the collection of material for direct examination and culture of Tritrichomonas foetus.     

The origin and species of yeasts in commercial preparations of bovine semen.

Whereas yeasts were not normally isolated from raw semen samples 13% of commercial frozen semen samples and 71% of preputial washings contained yeasts. Nine genera and 25 species of yeasts have been identified from these two sources. Yeasts originating in the preputial cavity were generally saprobic members of the genera Candida, Cryptococcus, Rhodotorula, Saccharomyces, Torulopsis and Trichosporon. Those originating as contaminants during processing were more likely to be opportunistic pathogens of the genus Candida. Conception was not necessarily affected by the presence of large numbers of Candida krusei or C. macedoniensis in the uterus.     

Measurement of immunoglobulins in reproductive tract fluids of bulls

Seminal, seminal vesicular, urethral and preputial fluids from bulls of two different age groups were assessed for quantitative differences in immunoglobulins. Selected markers were measured in individual samples to differentiate locally derived immunoglobulins from those present as a result of trauma or secretions from other accessory glands.Immunoglobulin levels in reproductive tract fluids from older bulls (5–6 years) were higher than those of younger bulls (3–4 years) and preputial fluids contained the highest concentration of immunoglobulins of all fluids examined. Similarities existed, however, among all fluids in the relative concentrations of immunoglobulins. IgG was generally in highest concentration, though the predominant subclass varied. A marked predominance of IgG₂ over IgG₁ occurred in preputial fluid samples of older bulls. IgA was in second highest concentration, and levels were often equal to or greater than those in serum. IgM was in low concentration and occasionally undetectable. IgG/IgA ratios did not exceed 5 in most of the reproductive fluids, whereas serum ratios were usually over 100. Proportional contents of albumin and immunoglobulin in reproductive tract fluids by comparison with those in serum indicated that substantial quantities of IgG as well as IgA were synthesized locally or derived by selective transport. Increased numbers of plasma cells in the lamina propria of the preputial and penile mucosa of older bulls were correlated with higher immunoglobulin concentrations in preputial fluid from older bulls, suggesting that differences in local synthesis were responsible.     

Intrapreputial infection of young bulls with bovine herpesvirus type 1.2 (BHV-1.2): acute balanoposthitis, latent infection and detection of viral DNA in regional neural and non-neural tissues 50 days after experimental reactivation

Venereal infection of bulls with bovine herpesvirus type 1.2 (BHV-1.2) may result in acute balanoposthitis followed by the establishment of latent infection, presumably in dorsal root nerve ganglia. We herein report the characterization of the acute and latent infection of young bulls with a Brazilian BHV-1.2 isolate and the investigation of neural and non-neural sites in which viral DNA persists during latent infection, i.e. 110 days after inoculation and 50 days after experimental reactivation. Intrapreputial inoculation of BHV-1.2 isolate SV-56/90 (10(6.5)pfu per animal) resulted in severe balanoposthitis, characterized by redness of the penis and preputial mucosa, coalescent vesicles and fibrinous exsudate in all four infected bulls. Virus shedding was detected in preputial secretions and semen up to days 14 and 13 pi, respectively. Dexamethasone administration at day 60 pi led to reactivation of the infection in all animals, resulting in virus shedding in preputial secretions and/or in semen. At day 50 post-reactivation (pr), the animals were euthanized and regional tissues were collected for PCR and virus isolation. Viral DNA was consistently detected in the dorsal root ganglia of nerves genito-femoral (4/4) and obturator (4/4); frequently in the pudendal (3/4), sciatic (3/4) and rectal caudal nerve ganglia (2/3). In addition, viral DNA was detected in the pelvic sympathetic plexus of one bull and in regional lymph nodes (deep inguinal (2/4); sacral (1/4); medial iliac (1/4)) of two bulls. No infectious virus could be recovered from homogenates of DNA positive tissues, indicating the absence of actively replicating virus. These results demonstrate that BHV-1.2 DNA may persist in several sacral nerve ganglia and in regional lymph nodes as well during latent infection, i.e. 50 days after experimental reactivation. These findings may help in understanding the pathogenesis of acute and latent genital infection by BHV-1.2.     

Experimental infections with Campylobacter fetus in bulls of different ages

Factors affecting the establishment of a carrier state of Campylobactor fetus subsp. fetus (venerealis) in preputial cavities of bulls were investigated by following infection in bulls of two different age groups until slaughter, 9–18 weeks post infection. Each of the four older bulls (66–74 months at infection) and three of the four younger bulls (41–49 months at infection) were culturally positive until slaughter, indicating no increased susceptibility with age. Relative proportions of immunoglobulin classes and albumin in preputial fluids were generally similar to those determined prior to infection, although protein concentrations decreased and ratios of IgG/IgA and IgG₁/IgG₂ increased in most of the bulls. An unexplained diminution of sample volumes occurred with progressive sampling. Changes in concentrations of proteins and in sample volumes bore no apparent relationship to infection of bulls with C. fetus. Low levels of C. fetus agglutinins were detected in sample obtained both before and after infection, but no appreciable rise in antibody titers occurred following infection. Alterations in superficial antigens of C. fetus isolates obtained during the course of infection were demonstrated in the majority of animals. The capacity of the organism to undergo antigenic variation and to provoke a minimal immune response may contribute to its prolonged survival in the preputial cavity.     

Surgical management of preputial injuries in bulls: 51 cases (1986-1994).

The purpose of this study was to compare success rates for preputial surgery depending on the initial diagnosis, ability to extend the penis, use of sedation and local anesthesia versus general anesthesia for surgery, and surgical technique. Medical records of 51 bulls treated surgically for preputial injury were reviewed. The mean age of the bulls was 2.5 years ranging from 1 to 5 years. Bos taurus breeds (82.3%) were more often affected than Bos indicus breeds (17.7%). The most common breeds represented in this study were Angus (45.1%), Simmental (11.8%), and brangus (9.8%). The seasonal incidence of preputial injuries was higher during the period of May-July (52.9%) and November-February (33.3%). The overall success rate was 70%. Posthioplasty was more successful than circumcision (90% to 43%) (P < 0.05). The success rate for surgeries performed under inhalation anesthesia in the surgery suite was 100% compared with a success rate of 63% for those animals operated on with injectable and local anesthesia in a rotary chute (P < 0.05). The success rate was 88% if the penis could be extended before the surgery and 36% if extension was not achieved (P < 0.05).     

Measurement of antibody in serum and genital fluids of bulls by ELISA after vaccination and challenge with Tritrichomonas foetus

An enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibody to Tritrichomonas foetus using both whole cell antigen (WCA) and membrane protein antigen (MPA). The test was used to detect specific antibody in serum, preputial washings and seminal plasma samples from 7 adult bulls which were vaccinated subcutaneously on 3 occasions with a membrane protein vaccine against T. foetus var brisbane in an oil adjuvant, and from 4 unvaccinated control animals. One month after administration of the third dose of vaccine, vaccinated and control bulls were repeatedly challenged with the live vaccine strain of the T. foetus. A steady increase in serum antibody titre was detected after each inoculation of vaccine when both antigens were used in the ELISA. However, MPA was more sensitive. After challenge, vaccinated bulls developed an increased titre. No specific antibody was detected in control bulls, except in one bull after challenge in which seroconversion was detected. The serum antibody titres of both groups of animals were also measured with the microagglutination test which proved less sensitive than the ELISA. Antibody titres to both antigens, although lower than in serum, were detected in the seminal plasma of vaccinated animals. The control bulls remained non-responsive. No antibody was detected by ELISA in preputial washings from either control or vaccinated bulls prior to challenge. Post-challenge, some of the vaccinated bulls were responsive with both antigens whereas the control bulls remained negative.     

Ulcerative posthitis in bulls in Uruguay.

Ulcerative posthitis in bulls in Uruguay is described; 1096 bulls in 17 different establishments were examined. The incidence of bulls grazing on improved pasture was 32.9% in those under one year and 80.8% in animals of 18 to 36 months of age. Bulls grazing on natural pasture showed an incidence of 63.5% in adult animals irrespective of age. Histologic characteristics of the preputial lesions were acanthosis, parakeratosis and hyperkeratosis followed by invasion of leukocytes in the epithelium and ulceration. Corynebacterium renale was isolated from 59 of 204 prepuces. Based on the isolation of C. renale, histological lesions and the greater incidence in animals on high planes of nutrition, it is postulated that the lesion is due primarily to production of ammonia following the hydrolization of urea by the organism.     

Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep

Histophilus ovis and Actinobacillus seminis were isolated from the preputial cavity of 6-month-old rams and the vagina of 6-month-old ewes at a substantially higher rate than that in mature (greater than 2 years old) rams and ewes. These organisms appeared to be a transitory component of the ovine genital flora, the prevalence of which was associated with age regardless of gender. Additional evaluation of the recoverability of H ovis and A seminis from the preputial cavity of rams from birth to 1 year of age indicated that the isolation rate from rams and predominance of the organisms in the preputial cavity differed greatly over this age period. These organisms were not recoverable until ram lambs were 12 weeks of age and were most prevalent at 20 weeks of age, after which recoverability of H ovis and A seminis from the preputial cavity steadily decreased, continuing through the time of the last evaluation at 1 year of age. The time period with which these organisms can be isolated from the preputial cavity is closely correlated with the time period when epididymitis associated with these organisms develops, and may be an important factor in the pathogenesis of epididymitis.     

Some Studies of Infectious Bovine Rhinotracheitis (IBR) Virus Infection in Calves

Six dairy calves, six and one-half to nine months old, were exposed to a strain of infectious bovine rhinotracheitis (IBR) virus of bovine fetal origin by one of the various routes — nasal, vaginal, preputial or contact. Neither after initial exposure nor following challenge of their immunity did any of these animals manifest the IBR respiratory syndrome, although two of them (inoculated per vagina/prepuce) developed pustular vulvovaginitis or balanoposthitis. Also, one five-day old dairy calf which had received colostrum and milk of its IBR-immune dam, was inoculated intranasally with the same strain of IBR virus. This animal exhibited severe signs of IBR. The virus was recovered from all but three of the seven calves after initial exposure and from all but one animal following challenge of their immunity. Immune responses of these calves resembled those of adult cattle.     

Mycoplasmas recovered from bovine genitalia, aborted foetuses and placentas in the Republic of South Africa

A total of 917 Mycoplasma isolations were made from 4 092 specimens originating from 2 874 cattle in private herds and at AI stations. The percentages of isolations from the different sources were: cervico-vaginal mucus 14,6%, semen 43%, preputial wash 25%, foetuses 3,3% and placentas 15%. Mycoplasma bovigenitalium, the most common isolate, was recovered from 39% of males, 47% of females, 25% of foetuses and 11% of placentas. A wide spectrum of mycoplasmas was present, and varying combinations were common. The possible pathogenic significance of the isolates is discussed.     

Presurgical antiseptic efficacy of chlorhexidine diacetate and providone-iodine in the canine preputial cavity

Antiseptic flushing of the canine prepuce and its exclusion from the surgical field are recommended before abdominal surgery to reduce the risk of bacterial contamination. The authors cultured the preputial cavity of 60 dogs prior to and following flushing with 0.05% chlorhexidine diacetate, 1% povidone-iodine, or 0.9% saline control. Bacterial growth was evaluated using a semiquantitative method, and bacterial organisms were subsequently identified. There were no significant differences between povidone-iodine and the saline control in any of the variables assessed. Chlorhexidine resulted in a significant decrease in the proportion of positive postflush cultures compared with povidone-iodine. Although not significant, the difference in adverse reactions between povidone-iodine (25%) and chlorhexidine diacetate (5%) suggests clinical relevance. Based on the results of this study, a 2 min flush with 0.05% chlorhexidine diacetate is recommended for presurgical preparation of the preputial cavity.     

Creation of a Urethral Conduit from a Preputial Indirect Flap in a Dog with Perineal Hypospadias

Objective— To report a surgical technique for creation of a urethral conduit using a preputial indirect flap.
Study Design— Case report.
Animals— Male Doberman Pinscher (6 weeks old).
Methods— A urethral conduit was constructed using a multistage preputial indirect flap in a dog with severe perineal hypospadias.
Results— Successful transfer of a perineal located urethral meatus to a more anatomically normal inguinal location was achieved with a preputial flap. The procedure was performed to aid control of recurrent urinary tract infections after anal and urethral separation in a dog with imperforate anus, urethrorectal fistula, and proximal perineal hypospadias.
Conclusions— Creation of a urethral conduit from a tubed bipedicle flap using redundant preputial tissue eliminated fecal contamination of the urethral meatus, aiding control of urinary tract infection. A good functional and cosmetic outcome was achieved.
Clinical Relevance— This technique should be considered in aiding control and prevention of urinary tract infections in dogs with proximal perineal hypospadias where a preputial remnant exists.     

Subtotal penile amputation and preputial urethrostomy in a dog

CASE DESCRIPTION: A 6-year-old castrated male Shih Tzu was evaluated because of intermittent bleeding during urination. CLINICAL FINDINGS: Necrosis of the cranial portion of the penile shaft extended distally from the preputial fornix. Penile necrosis secondary to strangulation from paraphimosis was diagnosed. TREATMENT AND OUTCOME: A midline preputiotomy incision was used to expose the penile shaft; amputation was performed caudal to the preputial fornix. The terminal portion of the urethra was anastomosed to the preputial mucosa, which allowed the dog to urinate through the preputial orifice. CLINICAL RELEVANCE: Unlike the more conventional urethrostomy procedures performed in dogs, preputial urethrostomy eliminates the potential for local skin irritation during urination. Preputial urethrostomy is also easier to perform in those dogs in which penile amputation is required adjacent to the preputial fornix. A release incision cranial to the prepuce can be used to facilitate caudal displacement of the preputial mucosa, which facilitates urethral anastomosis to this structure. A midline preputiotomy incision provided excellent exposure of the penile shaft for this surgical procedure.     

Penile and preputial tumours in the horse: a retrospective study of 114 affected horses

REASONS FOR PERFORMING STUDY: Preputial and penile tumours are more common in horses than in other domestic animals, but no large surveys of male horses with tumours of the external genitalia are available. OBJECTIVE: To present a retrospective analysis of male horses with neoplasms of the external genitalia. METHODS: The penile and preputial tumours of 114 horses were evaluated. Data recorded included age, gelding or stallion and breed; type and site of lesion; involvement of regional lymph nodes; histopathology (including grading of squamous cell carcinoma); and results of radiographic examination of the thorax. RESULTS: Mean age of horses was 19.5 years with no apparent breed predilection. Common presenting clinical signs were irregularities (e.g. the presence of a mass and/or ulceration) on the integument of the penis and prepuce, and purulent or sanguineous discharge from preputial orifice. Squamous cell carcinoma (SCC) was the most prevalent neoplasm followed by papillomas and melanomas. A basal cell carcinoma, neurofibrosarcoma, adenocarcinoma or fibrosarcoma were each found on single horses. Squamous cell carcinomas with poor differentiation had a higher tendency to metastasise than did more differentiated tumours. CONCLUSIONS: Squamous cell carcinoma is the most common urogenital tumour of the male horse and occurs primarily in old horses. Horses with poorly differentiated SCCs tend to have a higher incidence of regional metastases. Pathology of lymph nodes, even when not palpably enlarged, is a valuable diagnostic exercise. Radiology of the thorax to detect lung metastases is of little value.     

Common metastatic carcinoma of California sea lions (Zalophus californianus): evidence of genital origin and association with novel gammaherpesvirus

Tissues from 10 adult California sea lions (Zalophus californianus, seven females and three males) that had metastatic carcinoma in sublumbar area lymph nodes were examined histologically. A distinctive epithelial proliferative lesion interpreted as intraepithelial neoplasia was found in genital tracts of all ten animals; in vagina (5/7), cervix (7/7), uterus (3/7), penis (3/3) and prepuce (3/3). Intraepithelial neoplasia closely resembled metastatic carcinomas and was directly contiguous with invasive carcinoma in one animal. Rare eosinophilic intranuclear inclusion bodies were found in penile and preputial intraepithelial neoplasia (one animal), cervical intraepithelial neoplasia (one animal), invasive cervical carcinoma (one animal) and metastatic carcinoma (two animals). Electron microscopic examination of tissues from two sea lions (one with intraepithelial neoplasia and one with metastatic carcinoma) demonstrated viral particles consistent with a herpesvirus. An immunohistochemical stain for the latent membrane protein of Epstein-Barr virus was positive in intraepithelial neoplasia in one sea lion. Herpesvirus DNA sequences were detected by consensus primer polymerase chain reaction (PCR) in metastatic carcinomas from all four sea lions from which unfixed tumor samples were available. Results of sequencing were consistent with a novel gammaherpesvirus in the genus Rhadinovirus. DNA extracted from the four metastatic carcinomas also was tested for papillomavirus by Southern blot and PCR with consensus papillomavirus primers; all samples were negative by both methods. These findings support the genital origin of the sea lion carcinoma and implicate a novel gammaherpesvirus as a possible cause.     

Comparison of three sampling methods for the diagnosis of genital vibriosis in the bull

Three different methods of collecting preputial material for bacteriological examination were compared using 3 bulls infected with Campylobacter fetus subsp. fetus. The first method utilised a specially designed instrument to scrape the preputial and penile mucosa, int he second method plastic pipettes were used to aspirate material and the third method involved washing the preputial cavity with sterile peptone water. Bacteriological examination of the samples showed conclusively that scraping was the method of choice because more C. fetus positive samples were identified and there was less interference from contaminating organisms.