Estimation of receiver-operating characteristic curves to determine accuracy of a competitive enzyme-linked immunosorbent assay for the serodiagnosis of Brucella infection in domestic water buffalo (Bubalus bubalis) and cattle

OBJECTIVE: To estimate receiver-operating characteristic (ROC) curves for a competitive ELISA (c-ELISA) that is used in serodiagnosis of brucellosis in water buffalo and cattle, to determine the most appropriate positive cutoff value for the c-ELISA in confirmation of infection, and to evaluate species differences in c-ELISA function. SAMPLE POPULATION: Sera from 4 herds of cattle (n = 391) and 4 herds of water buffalo (381). PROCEDURE: Serum samples were evaluated for Brucella-specific antibodies by use of a c-ELISA. On the basis of previous serologic test results, iterative simulation modeling was used to classify animals as positive or negative for Brucella infection without the use of a gold standard. Accuracy of c-ELISA for diagnosis of infection was compared between cattle and water buffalo by comparison of areas under ROC curves. RESULTS: A positive cutoff value of 30% inhibition for c-ELISA yielded sensitivity and specificity estimates, respectively, of 83.9 and 92.6% for cattle and 91.4 and 95.4% for water buffalo. A positive cutoff value of 35% inhibition yielded sensitivity and specificity estimates, respectively, of 83.9 and 96.2% for cattle and 88.0 and 974% for water buffalo. Areas under ROC curves were 0.94 and 0.98 for cattle and water buffalo, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: ROC curves can be estimated by use of iterative simulation methods to determine optimal cutoff values for diagnostic tests with quantitative outcomes. A cutoff value of 35% inhibition for the c-ELISA was found to be most appropriate for confirmation of Brucella infection in cattle and water buffalo.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Conception rates in early postpartum ewes bred naturally or by intrauterine insemination

Ewes were treated with a medroxyprogesterone acetate (MAP) sponge for 8 d followed, at sponge removal, with 500 IU pregnant mare serum gonadotropin (PMSG) at d 30, 40 or 50 (d 0 = lambing) to induce estrus. Dry and lactating ewes were divided into equal numbers at each postpartum day and bred at estrus. Conception rates and number of accessory sperm were determined by flushing the oviducts 3 d after mating and examining the recovered ova. There was no effect (P greater than .05) of lactational status on conception rates. Conception rates increased (P less than .05) from d 30 (10%) to d 40 (45%) and from d 40 to d 50 (80%). There were fewer (P less than .05) ova with accessory sperm (5/26) in d-30 ewes compared with d-40 (10/27) or d-50 (12/24) ewes. In Exp. 2, ewes were assigned to two groups after receiving PMSG on d 30: 1) mated naturally or 2) inseminated during laparotomy near the uterotubal junction (UTJ). Dry and lactating ewes were divided evenly within each of the two treatments. Oviducts were flushed and ova were examined for cleavage. The conception rate was 60% in ewes that were inseminated in the UTJ vs 10% in ewes mated to rams (P less than .05). Lactational status had no effect on results. In conclusion, conception rates in postpartum ewes treated with MAP sponge and PMSG increased from postpartum d 30 to d 50 with natural breeding, and d-30 conception rates were increased over natural mating by insemination into the uterine horn near the UTJ.     

The prevalence and economic significance of Fasciola gigantica and Stilesia hepatica in slaughtered animals in the semi-arid coastal Kenya

Fasciola gigantica and Stilesia hepatica constrain ruminant productivity. The prevalence and economic losses caused by F. gigantica and S. hepatica in the ruminant production systems of Taveta division of Kenya were estimated in a retrospective appraisal of the slaughter records on the total number of animals slaughtered and livers condemned over the period 1989 to 2004. Only liver condemnations based on infestation by F. gigantica and S. hepatica were considered for purposes of this study. Liver condemnation rates differed significantly between bovines, caprines and ovines (p≤ 0.05) for F. gigantica (26%, 6.6% and 5.2%, respectively) and for S. hepatica (0.4%, 22% and 28%, respectively). The total loss through condemnation of both F. gigantica and S. hepatica infested livers was 4 408 272 KES (Kenyan shillings) (US$ 72 272). The proportion of loss in bovines, caprines and ovines, was 76%, 17% and 7%, respectively. Fasciolosis contributed 3 505 410 KES (79.5%) and S. hepatica infestation 903 210 KES (20.5%) to the total losses due to liver condemnations. The authors feel strongly that there is a need for more work on the two parasites using live animals to determine the prevalence in animals of various ages, species and breeds and the economics of disease control at farm level.     

Analysis of Haemophilus parasuis isolates from southern Ontario swine by restriction endonuclease fingerprinting.

To study the occurrence and distribution of various strains of Haemophilus parasuis in southern Ontario swine, organisms isolated from healthy swine, from specific pathogen-free and conventional herds, and from disease cases were examined using restriction endonuclease fingerprinting analysis. In most herds, several strains of H. parasuis could be detected although one or two strains usually predominated. Individual animals were colonized by a single or limited number of strains. In several cases, the same strains were isolated from more than one specific pathogen-free herd. Conventional herds carried a more heterogeneous population of H. parasuis. Only one strain was isolated which was common to more than one conventional herd. No strains were isolated which were common to both specific pathogen-free and conventional herds. None of the strains isolated from disease cases were found in healthy conventional or specific pathogen-free swine examined in this study.     

Evaluation of success rate of laparoscopic castration without orchidectomy in 32 mature horses

The surgical success of laparoscopic castration without orchidectomy results from avascular necrosis of the testes. However, failures and subsequent production of testosterone and stallion‐like behaviour have been previously identified. Laparoscopic castration without removal of the testes was performed in 32 horses with 2 normal descended scrotal testes between July 2006 and October 2012. The objectives of our study were to evaluate the success rate of laparoscopic castration without orchidectomy on descended testes in our population and assess complications and recovery time. Endocrine tests were performed after surgery and owners also asked to report on their horse's behaviour and ability to resume exercise. Castration was deemed successful if endocrine test results were in accordance with gelding values. Failed horses were castrated using a conventional technique and histology performed when possible. Basal testosterone levels decreased below 3 nmol/l after surgery in 30/32 cases. However, in 2 horses, stallion behaviour persisted and histological evaluation after inguinal castration showed residual viable tissue. Two horses maintained high testosterone levels after surgery associated with stallion‐like behaviour. Based on these results, 12.5% of horses (4/32 horses) failed to be considered a gelding based on either endocrine tests and/or histology. All horses, with one exception, were able to resume exercise less than a week after the procedure. Laparoscopic castration without orchidectomy appears to be a safe procedure. When assessing the success of laparoscopic castration, our total failure rate was 12.5% (4/32 horses) which is significantly higher than previously reported.     

Tenosynovitis of the digital flexor tendon sheath and annular ligament constriction syndrome caused by longitudinal tears in the deep digital flexor tendon: a clinical and surgical report of 17 cases in warmblood horses

REASONS FOR PERFORMING STUDY: Inflammation of the digital flexor tendon sheaths is a chronic and nebulous condition often unresponsive to medical and surgical treatment. OBJECTIVES: To evaluate the incidence of longitudinal tears (LT) as the underlying cause of chronic tenosynovitis and annular ligament constriction syndrome (ALCS) in warmblood horses. METHODS: The records of 25 horses with chronic tenosynovitis and ALCS in which tenoscopical inspection of the digital flexor tendon sheath (DFTS) was performed between 1999-2000 were reviewed. Of 25 horses, 17 were diagnosed with an LT in the deep digital flexor tendon (DDFT). All horses had a history of distension of the DFTS and/or signs of an ALCS. All cases presented typical signs of a chronic inflamed DFTS on ultrasonography and 11 horses showed ultrasonographic changes (echogenic material or an irregular outlining) at the lateral or medial border of the DDFT. The diagnosis of LTs of the DDFT was established in all cases by tenoscopy. Surgical treatment consisted of removal of the torn collagen fibrils using a mechanical resector and decompressing the fetlock canal by a transection of the palmar annular ligament (PAL) using a hook knife under tenoscopic control. RESULTS: Ten horses became sound and resumed their previous level of work, 3 horses remained lame, 4 horses returned to previous level of work but needed intrasynovial treatment of the DFTS and reduced competition frequency to remain sound. CONCLUSIONS: Horses presented with chronic inflamed DFTS and/or ALCS might suffer from LTs in the DDFT; however, the diagnosis cannot be established with absolute certainty using only ultrasonography. Longitudinal tears should be suspected if ultrasonographic changes are present lateral or medial to the border of the DDFT but tenoscopical examination of the tendon sheath is essential to establish an accurate diagnosis and an effective treatment. POTENTIAL RELEVANCE: The presence of these LTs might explain why some cases of chronic tenosynovitis of the digital flexor tendon sheath and/or ALCS do not respond on surgical transection of the PAL alone without tenoscopy.     

Glasser's disease and prevalence of subclinical infection with Haemophilus parasuis in swine in southern Ontario

Glasser's disease accounted for less than 1% of total swine mortalities in an 11 year retrospective postmortem survey of swine submissions at three provincial government diagnostic laboratories in southern Ontario. However, Glasser's disease was suspected in 17 of 83 boar mortalities at the Record of Production Boar Test Station between 1983 and 1985 and was much more common in specific-pathogen-free (SPF) boars than in conventional boars. The prevalence of the causative organism, Haemophilus parasuis, was determined for 19 SPF herds in Ontario classified as "Excellent" under the Ontario Swine Herd Health Policy. Nasal swabs from two-month-old pigs were cultured on chocolate agar containing 1.5 mug/mL lincomycin, 5 mug/mL bacitracin, and 0.1 mug/mL crystal violet. Three herds were negative for H. parasuis infection; 16 herds contained clinically healthy carrier pigs.