Molecular characterisation of Mycoplasma species isolated from the genital tract of Dorper sheep in South Africa.

Biochemical and molecular analysis were conducted on 34 strains of Mycoplasma species isolated between 2003 and 2009 from the genital tract of clinically healthy Dorper sheep and sheep with ulcerative vulvitis and balanitis. Earlier publications identified the causative agent as Mycoplasma mycoides mycoides large colony (MmmLC) and Arcanobacterium pyogenes. The aims of the study were to characterise Mycoplasma species isolated from the genital tract of Dorper sheep with polymerase chain reaction assay, cloning and gene sequencing. Basic Local Alignment Search Tool (BLAST) results revealed six predominant Mycoplasma species: Mycoplasma arginini, Mycoplasma bovigenitalium, Arcanobacterium laidlawii, MmmLC, Mycoplasma sp. ovine/caprine serogroup II and M. canadense. Sequencing of the 34 isolates were analysed using phylogenetic methods, and 18 (50%) were identified as M. arginini with 99% - 100% similarity to M. arginini from England and Sweden. Six isolates showed 99% similarity to M. bovigenitalium strains from Turkey and Germany. Two isolates had 99% similarity to an M. sp. ovine/caprine sero group II from the United Kingdom. BLAST for two isolates revealed 99% similarity to Acholeplasma laidlawii from India, another two were 99% similar to MmmLC strain from Sweden, two showed 98% similarity to Mycoplasma sp. Usp 120 from Brazil, and two isolates have a 97% - 99% similarity to M. mm. Jcv1 strain from the United States of America. Finally, one isolate showed similarity of 99% to Mycoplasma canadense strain from Italy. The findings support the hypothesis that ulcerative vulvitis and balanitis of Dorper sheep in South Africa (SA) is a multifactorial disease with involvement of different Mycoplasma species.

Genome-wide association study for posthitis in the free-living population of European bison (Bison bonasus).

BACKGROUND: About 5-6% of the European bison (Bison bonasus) males are affected by posthitis (necrotic inflammation of the prepuce) and die in the wild forest. Despite many years of study, pathogenesis of this disease has not yet been determined. The main aim of the study was to find SNP markers significantly associated with the incidence of posthitis and mine the genome for candidate genes potentially involved in the development of the disease. RESULTS: It was shown that relatively small number of SNPs effects reached genome-wide significance after false discovery rate (FDR) correction. Among 25 significant markers, the highest effects were found for two SNPs (rs110456748 and rs136792896) located at the distance of 23846 bp and 37742 bp, respectively, from OR10A3 gene (olfactory receptor genes), known to be involved in atopic dermatitis in humans. It was also observed that five other significant SNP markers were located in the proximity of candidate genes involved in severe diseases of skin tissue and cancer/tumour development of epithelial or testicular germ cells, which suggest their potential participation in the posthitis. The 25 investigated SNPs showed marked differences in allelic and genotypic frequencies between the healthy and affected bison groups. CONCLUSIONS: The 2 Mb region of the BTA15 chromosome is involved in genetic background of posthitis and should be closer examined to find causal mutations helpful in better understanding of the disease ethology and to control its incidence in the future.

Factors associated with failure in breeding soundness examination of Western USA rams.

Breeding-soundness examination (BSE) and eradication of Brucella ovis infection in rams are critical components of flock-health programs. The aims of this retrospective, cross-sectional study were to describe the results of BSE in a large sample of rams in the Western USA and to determine the association between BSE outcome and the semen collection method (penis manually extended vs. retained in the preputial cavity), ram body-condition score (BCS), the presence of ulcerative posthitis, and the size of the flock of origin. We evaluated the first BSE in a given year for rams from Colorado, Wyoming, and Utah, USA, from 2000 through 2007. Breeding-soundness examination consisted of physical examination, scrotal circumference and BCS measurement, semen collection by electroejaculation, and microscopic examination of semen motility, morphology, and leukocyte concentration. We assigned a reason for failure to each failed BSE and used multivariable logistic and Poisson regressions to measure associations between ram and flock variables and the risk or reason for failure on BSE. A non-random, owner-selected subset of rams was tested for antibodies to B. ovis by serum indirect ELISA (iELISA). The Rogan-Gladen corrected B. ovis seroprevalence was measured. Of the 14,667 BSEs performed on 11,804 rams, 29.0% were classified as "failed;" the most common reason for failure was substandard semen parameters (43.8%). Breeding-soundness examinations were more likely to have been categorized as failure for inflammatory causes when performed on rams from medium-sized flocks (OR 1.6; 95% CI 1.1, 2.3) and large flocks (OR 1.4; 95% CI 1.0, 1.9) (P=0.02), suggesting that larger flocks are at higher risk of contagious diseases. The adjusted seroprevalence of B. ovis antibodies among tested rams in this study was 10.0%. Of 233 rams seropositive to B. ovis, 125 (53.6%) were subclinical, a finding that supports the importance of this test in ram BSE. We found that emaciation in rams was associated with an increased risk of BSE failure from substandard semen parameters (P<0.001), but ulcerative posthitis and the semen collection method were not (P=0.09 and 0.34, respectively). However, collection of semen with the penis retained in the preputial cavity resulted in greater odds of leukospermia relative to semen collection with the penis extended (OR 4.1; 95% CI 2.9, 5.9; P<0.001), presumably from contamination of the semen sample with preputial leukocytes. For ram BSE, therefore, semen collection with the penis manually extended from the sheath is recommended to limit leukocyte contamination of the sample.

Balanoposthitis, dyspareunia, and treponema in the critically endangered Gilbert's potoroo (Potorous gilbertii).

The Gilbert's potoroo (Potorous gilbertii) is one of Australia's most critically endangered mammals with a current estimated population of 70 individuals. Both the wild and captive populations have a long history of balanoposthitis with associated crusting, ulceration, and preputial discharge. We sought to identify the microbial species found in the discharge, determine their significance in causing balanoposthitis, and correlate these findings with reproductive success and survivorship. Bacteriologic examination revealed the discharge to be a polymicrobial infection involving Treponema spp., Actinobacillus spp., and Pasteurella spp. Preputial histopathology reported a moderate, chronic, erosive inflammatory response with diffuse, moderate to marked secondary epithelial hyperplasia in conjunction with moderate numbers of spirochetes, suggesting a causative relationship. Clinical examination, preputial biopsies, and serologic screening found no evidence of associated systemic disease. The clinical investigation of Treponema is significant with respect to the overall recovery of Gilbert's potoroo, given the clinical and histopathologic similarities to Treponema paraluis-cuniculi found in rabbits, causing dyspareunia, and the severity of the associated balanoposthitis.

Chronic prostatitis, cystitis, pyelonephritis, and balanoposthitis in a cat.

An adult, intact male domestic shorthair presented for preputial swelling and urinary incontinence. A caudal abdominal mass was palpated. A transabdominal ultrasound examination showed severe prostatomegaly with abnormal tissue extending along the urethra. The cat was euthanized due to the owner's financial constraints and the veterinarians' suspicion of a poor long-term prognosis. Biopsies showed chronic active inflammation of the prostate, bladder, kidneys, ureters, penis, and prepuce most consistent with a chronic infectious process. Reports of feline prostatic disease of any kind are rare. Chronic prostatitis may have a more favorable prognosis than feline prostatic adenocarcinoma, currently the most commonly reported disease of the feline prostate.

Ulcerative vulvitis and balanitis in sheep flocks.

Outbreaks of ulcerative vulvitis and balanitis occurred in three commercial sheep flocks in England and Wales. Between 29 and 44 per cent of the ewes were affected; most of the lesions resolved in three weeks. Pathogens such as mycoplasmas, which have previously been associated with these conditions, were not detected despite using improved laboratory techniques. In one of the flocks, ovine herpesvirus type 2 was detected by pcr in the blood of two acutely affected ewes, from the vulval ulcers of one of them, and from the penis of an affected ram.

High prevalence of ulcerative posthitis in Rasa Aragonesa rams associated with a legume-rich diet.

This study describes an outbreak of ulcerative posthitis that eventually affected 76 of 80 of rams in a flock of Rasa Aragonesa sheep on a legume-rich diet. Lesions were confined to the prepuce and varied from mild hyperaemia to ulcerations. Corynebacterium renale was isolated from the lesions. Treatment of an initial group of 17 was based on a change of diet and topical treatment with Veterin Banedif with prednisolone once a day for 15 days and was successful. Lesions in this group were completely resolved after 15 days. The relationship between a legume-rich diet, ruminal alkalosis, elevated urine pH and posthitis is discussed.

Ulcerative balanitis and vulvitis of Dorper sheep in South Africa: a study on its aetiology and clinical features.

Ovine ulcerative balanitis and vulvitis in sheep of the Dorper breed has been observed in South Africa since 1979. Its aetiology has not been conclusively resolved, and there is some discrepancy in descriptions of its clinical features. In order to identify the pathogenic micro-organism/s that contribute to the occurrence of the disease, the microflora in the genital tracts of both clinically healthy and affected sheep were isolated and compared. Bacteriological examination of materials from affected and unaffected sheep resulted in the isolation of Arcanobacterium pyogenes from 44.2% and 17.2% of them respectively. This difference is statistically significant (P < 0.01). Seventy-four per cent of the isolates originated from severe clinical cases. Mycoplasmas were isolated from 49.3% of 116 clinically normal sheep and 78.2% of 104 affected sheep. There were significant differences in their rates of isolation in clinical groups (P < 0.05). Of all the mycoplasma isolates, Mycoplasma mycoides mycoides large colony variant (MmmLC) was isolated from 61.5% of clinically diseased sheep while 6.0% of the isolates were from apparently healthy animals (P < 0.05). The study threw light on the prevalence of mycoplasmas in the genital tract of apparently healthy sheep and, at the same time the identity of the mycoplasma pathogen associated with ulcerative balanitis and vulvitis was revealed. The findings of this investigation therefore confirmed the involvement of mycoplasma, particularly that of MmmLC large colony, in the disease in Dorper sheep in South Africa, and it was concluded that this microorganism is an important pathogen of balanitis and vulvitis in them. The study furthermore demonstrated a probable synergism between A. pyogenes and MmmLC. Finding these 2 organisms together occurred 53.4 times more frequently in the affected sheep than in the unaffected, which emphasises the probable multifactorial nature of the disease. The association between age and the presence of clinical signs was statistically significant. It was found that young sheep were more likely to have lesions than adult sheep. Clinical observations showed that the typical ulceration appears to be confined to the glans penis and lips of the vulva; no ulceration was observed on the shaft of the penis and prepuce or vaginal vestibule. In uncomplicated cases inflammation of the prepuce and vaginal vestibule is not a regular feature of the disease. Therefore the names ulcerative balanitis and vulvitis most accurately describe the nature of the disease in South Africa.

In vitro antimicrobial susceptibility of Mycoplasma mycoides mycoides large colony and Arcanobacterium pyogenes isolated from clinical cases of ulcerative balanitis and vulvitis in Dorper sheep in South Africa.

The in vitro activities of enrofloxacin, florfenicol, oxytetracycline and spiramycin were determined against field isolates of Mycoplasma mycoides mycoides large colony (MmmLC) by means of the broth microdilution technique. The minimum inhibitory concentrations (MICs) of these antimicrobial drugs were determined for a representative number of 10 isolates and 1 type strain. The susceptibility of Arcanobacterium pyogenes to enrofloxacin, oxytetracycline and tilmicosin was determined by means of an agar disk diffusion test. The MICs of enrofloxacin, florfenicol, oxytetracycline and spiramycin were within the ranges of 0.125-0.5, 1.0-2.0, 2.0-4.0 and 4.0-8.0 microg/ml, respectively. This study has shown that resistance of MmmLC against enrofloxacin, florfenicol, oxytetracycline and spiramycin was negligible. All the field strains of A. pyogenes that were tested were susceptible to enrofloxacin, oxytetracycline and tilmicosin with mean inhibition zones of 30.6, 42.3 and 35.8 mm, respectively. Although there is lack of data on in vivo efficacy and in vitro MIC or inhibition zone diameter breakpoints of these antimicrobial drugs for MmmLC, the MIC results indicate that these 4 classes of antimicrobial drugs should be effective in the treatment of ulcerative balanitis and vulvitis in sheep in South Africa.

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.

Virologic investigations of free-living European bison (Bison bonasus) from the Bialowieza Primeval Forest, Poland.

We conducted virologic investigations on postmortem specimens from 261 free-living European bison (Bison bonasus) from the Bialowieza Primeval Forest, Poland collected between 1990 and 2000. Fifty-four of 94 males had balanoposthitis; none of the 167 female bison examined had reproductive tract lesions. Peripheral blood, swabs, and various tissues were analyzed for bovine viruses as well as for viral DNA by bovine herpesvirus 1 (BoHV-1) and bovine herpesvirus 4 (BoHV-4) specific polymerase chain reaction (PCR) technique. An infectious bovine rhinotracheitis like BoHV-1 strain was isolated from the spleen of a female bison calf and additionally was detected by nested PCR from splenic tissue. None of the bison had significant antibody titers against BoHV-1, bovine herpesvirus 2, BoHV-4, caprine herpesvirus 1, cervid herpesvirus 1, or bovine viral diarrhea (BVD) virus-1. However, low antibody titers in two animals indicate that this European bison population has been exposed to BVD virus or BVD-like viruses and BoHV-2.

Antibodies against Mycoplasma bovigenitalium in free-living European bison (Bison bonasus) with balanoposthitis.

Since 1980 severe chronic balanoposthitis has been observed in free-living European bison (Bison bonasus) in the Bialowieza Primeval Forest (Poland). Sera of 50 bison with balanoposthitis and 48 clinically healthy male and 49 female bison were investigated for antibodies against Mycoplasma bovis and M. bovigenitalium by western blot analysis. Prevalence of antibodies against M. bovigenitalium was significantly higher in bison with balanoposthitis than in unaffected male bison. Mycoplasma bovigenitalium may play a role in the pathogenesis of balanoposthitis in European bison.

Contagious agalactia of small ruminants: current knowledge concerning epidemiology, diagnosis and control.

Contagious agalactia of small ruminants is a syndrome which principally affects the mammary glands, joints and eyes. The main causal agents are Mycoplasma agalactiae in sheep, and M. agalactiae, M. mycoides subsp. mycoides large colony type and M. capricolum subsp. capricolum in goats. In addition, M. putrefaciens can produce a similar clinical picture, particularly in goats. Contagious agalactia occurs on all five continents and is often enzootic. The evolution of the infection tends to be chronic in affected animals and herds. Symptomless shedding of mycoplasmas, mainly in the milk, may persist for a long time. These insidious infections, associated with carriage in the ears of healthy animals, are difficult to diagnose and to control. The main mode of transmission between flocks is related to the sale of carrier animals and contact during transhumance, whereas transmission within a flock occurs through contact, suckling and milking. This review discusses the clinical features, epidemiology, treatment, prevention and control of the disease.

Production, behaviour and fertility of merino wethers, hemi-castrates with reduced testicular parenchyma and induced cryptorchids.

Greasy wool production, body weight, behaviour and fertility of male, fine wool Merino sheep were studied from 10 to 21 months of age to assess their suitability for wool production. The males were either castrated (wethers), or were hemi-castrated with a vasectomy and reduction (either partially or in full) of the parenchyma of the retained testicle, or the testicles pushed up into the inguinal canal and the scrotum shortened (induced cryptorchids). There were significant differences among all groups in body weight, with increases being associated with the larger amounts of testicular tissue present. Fertility was low in the induced cryptorchids at 19 months of age. The hemi-castrates with reduced testicular parenchyma were infertile. The hemi-castrates with reduced testicular parenchyma had significantly heavier greasy fleece weights than the wethers and induced cryptorchids, which had similar fleece weights. The wether group suffered a higher prevalence of posthitis in spring and autumn than the hemi-castrate or induced cryptorchid groups. Those hemi-castrates with the largest amount of testicular parenchyma retained and the induced cryptorchids exhibited masculine behaviour patterns and had an increased incidence of fly strike to the head. The principle of hemi-castration and reduction of the testicular parenchyma is shown to be applicable in wool producing flocks with the potential to increase greasy wool production while minimising the management and marketing limitations previously associated with induced cryptorchids.

The identification of Mycoplasma mycoides mycoides LC as the aetiological agent of balanoposthitis and vulvovaginitis in sheep in South Africa.

Clinical ulcerative balanoposthitis and vulvovaginitis was experimentally reproduced in 14 sheep infected with a Mycoplasma mycoides mycoides LC field strain, isolated from the Straussheim Dorper stud. The study encompassed a series of field observations, a therapeutic trial and experimental investigations. A wide range of bacteria and various mycoplasma spp., but no viruses, were isolated from a large number of infected animals.

Balanoposthitis in bulls due to bovine herpesvirus in south Brazil.

We describe an acute outbreak of balanoposthitis in bulls at an artificial insemination station in South Brazil. Bovine herpesvirus was isolated from preputial swabs in the Crandell feline kidney cell line and secondary fetal bovine lung cells and identified using the fluorescent antibody technique and electron microscopy. Polyclonal antibodies against the whole virus were used for identification of the bovine herpesvirus with the fluorescent antibody technique. Herpesvirus particles could be seen in the infected cells by electron microscopy. Eleven bulls had clinical signs resembling balanoposthitis and nine yielded virus.

Balanoposthitis in bulls due to bovine herpervirus in South Brazil

We describe an acute outbreak of balanoposthitis in bulls at an artificial insemination station in South Brazil. Bovine herpesvirus was isolated from preputial swabs in the Crandell feline kidney cell line and secondary fetal bovine lung cells and identified using the fluorescent antibody technique and electron microscopy. Polyclonal antibodies against the whole virus were used for identification of the bovine herpesvirus with the fluorescent antibody technique. Herpesvirus particles could be seen in the infected cells by electron microscopy. Eleven bulls had clinical signs resembling balanoposthitis and nine yelded virus