Lack of Correlation Between Papillomaviral DNA in Surgical Margins and Recurrence of Equine Sarcoids

Bovine papillomavirus (BPV) types 1 (BPV-1) and 2 (BPV-2) are causally associated with the development of equine sarcoid tumors. Recurrence rates after surgical excision of sarcoids are estimated to be 30%–40%. We hypothesized that the presence of BPV DNA in histologically tumor-free surgical margins of sarcoids is associated with risk of recurrence, and increased quantity of BPV DNA is associated with increased risk of recurrence. Formalin-fixed sarcoids classified as “completely excised” histologically were obtained from two institutions. A total of 25 tumors were included, eight of which recurred within 1 year of excision. Qualitative and quantitative polymerase chain reaction (PCR) tests for detection of BPV-1 and BPV-2 were performed on neoplastic tissue and tumor-free surgical margins in formalin-fixed paraffin-embedded biopsy specimens following DNA extraction. Bovine papillomavirus-1 was found in all tumor samples and in histologically “clean” margins of 21 samples, whereas BPV-2 was found in only two tumor samples. Although quantitative PCR was more sensitive than qualitative PCR in detecting BPV DNA in surgical margins, there was no significant difference in the presence of BPV-1 or BPV-2 DNA in margins of tumors that recurred versus those that did not recur for either test. Although this study is limited by sample size, our results suggest that PCR analysis of surgical margins for BPV DNA is not a reliable method to predict equine sarcoid recurrence after resection.     

Effects of unilateral castration and unilateral cryptorchidism of the Holstein bull on plasma gonadotropins, testosterone and testis anatomy

The effects of unilateral castration (UC) and induced unilateral cryptorchidism (CR) on plasma hormones and testis anatomy were studied in 36 Holstein bulls altered at either 3, 6 or 9 mo of age (n = 12). Plasma hormone concentrations were determined in six samples collected at hourly intervals on d 0, 1, 3, 7, 14 and 30, and then at monthly intervals through 6 mo after gonadal manipulation. Although plasma testosterone (T) showed a transient decrease (P less than .05) immediately after treatment, mean plasma concentrations of luteinizing hormone (LH) and T were unaffected by UC or CR over the 6-mo period (P greater than .05). Both hormones increased (P less than .05) in concentration with advancing age. Plasma follicle stimulating hormone (FSH) concentration was greater (P less than .05) in UC than in intact (IN) bulls overall, while FSH in CR bulls did not differ (P greater than .05) from either group. At slaughter, 11 mo after gonadal alteration, mean testis weight, ratio of testis weight to body weight and mean testis sperm cell numbers were increased (P less than .05) in UC bulls compared with mean testis values in intact (IN) bulls. Unilateral castration increased (P less than .05) seminiferous tubuler diameter and seminiferous epithelial cell height from basement membrane to the border of the lumen, but did not alter the ratio of tubuler to interstitial space within the testis. Seminiferous tubuler diameter and epithelial cell height were increased (P less than .05) in CR compared with IN bulls. Unilateral gonadal alteration at 3 mo of age caused a greater (P less than .05) hypertrophy of the scrotal testis in both UC and CR bulls than alteration at 6 or 9 mo of age. Results indicate that unilateral gonadal disruption is followed by rapid compensation in testis T production, little change in systemic LH and a rapid increase in secretion of FSH in the bull within those ages investigated. Further, UC elicits a greater compensatory hypertrophy than CR and the pituitary-testis endocrine axis is more responsive to alteration at 3 mo than at 6 or 9 mo of age in the bull.     

Molecular characterisation of bovine faecal Escherichia coli shows persistence of defined ampicillin resistant strains and the presence of class 1 integrons on an organic beef farm

Antimicrobial use is heavily restricted on organic farms; however, few studies have been conducted to investigate the impact this has on the epidemiology of resistance in pathogenic and commensal bacteria. We investigated the persistence of antimicrobial resistant Escherichia coli within an organic beef herd over a period of 28 months. Faecal samples collected monthly from three calf cohorts and annually from adult cattle and environmental samples, were screened for the presence of ampicillin, apramycin and nalidixic acid resistant E. coli. The prevalence of ampicillin resistance ranged from 27.3 to 40.7% in the annual herd and environmental samplings (n=22-55) and was greater in the calf cohorts, with a peak cohort prevalence of >47% in all 3 years (n=16-18). Apramycin and nalidixic acid resistant E. coli were rare. Pulsed-field gel electrophoresis (PFGE) identified 10 main genotype groups within the herd, with evidence of strain transmission between different livestock groups, animal species and years. Multiple resistance was found in >44% of isolates tested, with ampicillin, neomycin, sulphamethoxazole and tetracycline carriage the commonest phenotype identified. PCR detected the presence of class 1 integrons in <5% of resistant isolates, 6/7 of which were of cattle origin. These data demonstrate that ampicillin resistant E. coli was common on the farm despite restricted antimicrobial use, although strain diversity was low. Persistence of defined genotype groups was observed between years, together with the transmission of resistant strains between different animal species on the farm.     

Limited morphological,physiological and genetic diversity of Phytophthora palmivora from cocoa in Papua New Guinea

In Papua New Guinea (PNG) cocoa (Theobroma cacao) is one of the most important cash crops grown in the tropical lowland and island regions. As in most cocoa‐growing areas, phytophthora black pod and canker cause significant yield losses. Cocoa breeding activities in PNG are focused in East New Britain province where disease control recommendations are also developed. This study tested the hypothesis that there was no diversity in the Phytophthora palmivora population causing black pod on cocoa by characterizing the variation in pathogen populations within and between the five major cocoa‐growing areas. Diseased pods were sampled hierarchically from the five locations and additional isolates were collected from soil, stem and leaf lesions, or retrieved from culture collections. Morphological characters showed continuous variation within the range described for P. palmivora. Genetic analysis revealed that the isolates belonged to one dominant clonal lineage, with restricted distributions of several other subpopulations. Lowest diversities were found in the geographically isolated Karkar Island and East Sepik province. Soil isolates showed greater genetic diversity than isolates from cocoa lesions. Intra‐farm variation was as much as inter‐farm or inter‐province variation. Both mating types were detected, although no strong evidence of sexual recombination was observed. The analysis revealed limited geographic, temporal or host specialization, suggesting continuous selection for pathogenicity from a genetic pool of P. palmivora. These findings have significant implications on the deployment of cocoa genotypes, enforcement of inter‐province quarantine and sustainable disease management strategies.     

Collection Rates and Morphology of Equine Oocytes Obtained from Immature Follicles after Treatment with Equine Pituitary Extract (EPE) and Human Chorionic Gonadotropin

This study examined the effect of treating mares with equine pituitary extract (EPE) in combination with human chorionic gonadotropin (hCG; EPE/hCG) on the recovery rate of immature oocytes by ovum pick-up (OPU) and on oocyte morphology. Ten mares were subjected to each of two treatments in a random sequence: superstimulated with EPE (25 mg, twice daily) and treated with hCG (2,500 IU) or control (no exogenous treatment). The cytoplasmic morphology of oocytes recovered was evaluated through transmission electron microscopy. Follicular fluid was collected at aspiration for progesterone analysis, which was performed by radioimmunoassay. The EPE/hCG did not increase the oocyte recovery rate from immature follicles when compared with the controls (15.5% and 16.7%, respectively). A significantly higher oocyte recovery rate per mare was observed (70% versus 50%). However, precocious granulosa cell expansion was observed with EPE/hCG treatment in contrast to the control (64.4% and 33% of follicles with expanded cumulus, respectively), and increased intrafollicular progesterone concentration was also seen (158.80 ng/mL versus 82.05 ng/mL). The ultrastructural analysis of oocytes from both groups showed morphologic features related to immaturity. Numerous vesicles containing cortical granules were found, distributed in clusters into the cytoplasm, and junctional complexes were still seen between oocyte and granulosa cells. In conclusion, EPE/hCG treatment induced some follicular modifications, but the recovery rate was not increased. All oocytes examined presented signs of immaturity.     

Surgical technique to cannulate the rete testis of the goat

A surgical technique was devised to collect rete testis fluid from 14 mature goats. The tubular portion of the extratesticular rete testis was cannulated where it penetrated the tunica albuginea of the testis under the center of the head of the epididymis. The tip of the cannula was sutured in place in the extratesticular rete testis and the opposite end was passed through the scrotum and into a plastic collection bottle. The bottle was glued to pillow ticking, which was sutured to the scrotum. Continuous flow of rete testis fluid occurred for 0 to 14 days and was usually terminated by a sperm clot in the cannula. The flow rate was 0.59 +/- 0.37 ml/hr.