Western blot detection of PrP Sc in archived paraffin-embedded brainstem from scrapie-affected sheep

Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies (TSE) or prion diseases. Immunoassays that identify disease-associated prion protein (PrP Sc) are integral to the diagnosis of scrapie and other prion diseases. Results obtained by either immunohistochemistry (IHC) or Western blot (WB) assay are generally adequate for the definitive diagnosis. Approved or accepted methods for WB diagnosis of TSEs requires the use of fresh or frozen nonfixed tissue samples, whereas formalin-fixed, paraffin-embedded tissue is required for the localization of PrP Sc by IHC. Because disparate processing methods are used for these accepted diagnostic techniques, separate tissue samples are collected from the same animal. Occasions arise in which there is either insufficient quantity of tissue available to complete analysis by both techniques or initial tissue processing is incompatible with one of the assays. Also, results between the assays may differ because of the vagaries of sampling, especially in case material that contains moderate-to-low levels of PrP Sc. The present article describes a method to conduct a WB assay from the same paraffin-embedded brainstem sample used for the IHC diagnosis of experimentally induced sheep scrapie.     

Cluster of cases of malignant schwannoma in cattle

Between 1998 and 2001, several cases of ataxia and paresis followed by recumbency and death were reported in cows from different farms in a restricted area of the Argentinian Patagonia. Five cases of this cluster were studied and a diagnosis of malignant schwannoma was established. Electron microscopy (em) of tumour samples from three of the animals revealed intracytoplasmic or interstitial structures resembling retroviral particles. Attempts to isolate a viral agent from the tumours were unsuccessful but the epidemiological data and the em findings suggest a viral aetiology.     

Relationship of ruminal temperature with parturition and estrus of beef cows

Spring-calving Angus cows (n = 30) were used to evaluate changes in ruminal temperature (RuT) related to parturition and estrus. Cows were synchronized and artificially inseminated with semen from a single sire. Temperature boluses were placed in the rumen at 7.0 ± 0.2 mo of gestation. Boluses were programmed to transmit RuT every 15 min. Cows (BW = 623 ± 44 kg, BCS = 4.9 ± 0.4) calved during 3 wk, and estrus was synchronized at 77 ± 7 d after calving with PGF(2α). Cows were observed every 12 h to detect estrus. Daily average ambient temperatures ranged from 2 to 22 °C during parturition (February to March) and 17 to 25 °C during estrus (May to June). Ruminal temperature from 7 d before to 3 d after parturition and 2 d before to 2 d after visual detection of estrus was analyzed using the MIXED procedure. Ruminal temperatures <37.72 °C were attributed to water consumption and excluded from analyses. Day did not influence (P = 0.36) RuT from d -2 to -7 before parturition (38.94 ± 0.05 °C). Ruminal temperature decreased (P < 0.001) from d -2 to d -1 before parturition (38.88 ± 0.05 to 38.55 ± 0.05 °C, respectively). Ruminal temperature was not influenced (P = 0.23) by day from 1 d before to 3 d after parturition (38.49 ± 0.05 °C). Ruminal temperature at 0 to 8 h after detection of estrus (38.98 ± 0.09 °C) was greater (P < 0.001) compared with RuT at the same daily hour of the day before (38.37 ± 0.11 °C) or the day after estrus (38.30 ± 0.09 °C). Ambient temperature did not influence (P > 0.30) RuT at parturition or estrus. Ruminal temperature decreased the day before parturition and increased at estrus in spring-calving beef cows and has potential use as a predictor of parturition and estrus.     

Taenia solium porcine cysticercosis: viability of cysticerci and persistency of antibodies and cysticercal antigens after treatment with oxfendazole

The aim of this study was to assess the effect of treating Taenia solium infected pigs with oxfendazole (OFZ) on viability and clearance of cysticerci and the corresponding persistence of specific antibody isotypes (IgG(total), IgG1, IgG2 and IgA) and circulating cysticercal antigen (CCA). Antibody isotypes and CCA responses were measured by antibody-ELISA (Ab-ELISA) and antigen ELISA (Ag-ELISA), respectively. Correlations were made between antibodies, CCA and the total number of cysticerci enumerated at necropsy. Forty pigs with cysticercosis were randomly allocated into two groups: Treatment group (n=20) was treated with OFZ at 30 mg/kg orally while the treatment control group (n=20) was not treated. Five uninfected pigs served as negative controls. Pigs were killed at 1, 4, 8 and 26 weeks post-treatment (wkpt). Overall, the mean total cyst count in treated pigs was 2904+/-5397 (mean+/-S.D.) while in the controls it was 6235+/-6705. Mean cyst viability was 5+/-11% (mean+/-S.D.) and 97+/-4% in treated and control pigs, respectively. Results showed that OFZ killed muscular cysticerci over a period of 4 weeks but failed to kill cerebral cysticerci. Antibodies, CCA responses and clearance of dead cysts from the meat, depended on the cyst intensity of individual pigs at time of treatment since both antibody and CCA correlated with intensity of cysticerci at necropsy (r=0.441, P=0.005; r=0.654, P<0.001), respectively. IgG1 responses were the best indicator of treatment efficacy because they were predominant in both infected treated and control pigs and disappeared early after treatment. Both Ab/Ag-ELISA failed to detect cysts in the brain. Though dead cysticerci took some time (26 wkpt) to clear from the meat, treatment of porcine cysticercosis with OFZ should, in combination with other intervention measures be considered as an important, cost-effective measure in the control of taeniosis/cysticercosis.     

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

This case report describes tuberculosis (TB) due to infection with Mycobacterium bovis (M. bovis) in alpaca (Lama pacos) on a farm in Ireland. Two severely debilitated alpaca were presented to the University Veterinary Hospital, University College Dublin in November 2004. Bloods were taken, and haematology and biochemistry results were indicative of chronic infection. Radiological examination showed evidence of diffuse granulomatous pneumonia suggestive of tuberculosis. On necropsy there were granulomatous lesions present throughout many body organs including lung, liver, kidney, intestine as well on peritoneum and mesentery. Culture of acid-fast bacilli from lesions led to a diagnosis of tuberculosis due to M. bovis. The use of intradermal skin testing proved inefficient and unreliable for ante mortem diagnosis of tuberculosis in alpaca. Infection due to M. bovis should be considered among the differential diagnoses of debilitating diseases in alpaca, particularly those farmed in areas known to be traditional black spots for tuberculosis in cattle.     

Humoral response to Mycobacterium avium subsp. avium in naturally infected ring-neck doves (Streptopelia risoria)

Creation of a reliable and easy to use serologic test would greatly improve ante mortem diagnosis of Mycobacterium avium subsp. avium and aid in the control of avian mycobacteriosis, particularly in captive birds. In order to determine whether serodiagnostics could be of value in testing ring-neck doves (Streptopelia risoria) for M. a. avium infection, Western blot analysis was used to assess the humoral response of ring-neck doves exposed to M. a. avium, and to evaluate whether an association could be made between the humoral response and necropsy findings, histopathology, culture, and PCR testing. Western blot results were examined for reactivity patterns associating humoral response with infection status, severity and type of lesions (diffuse vs. multifocal granulomatous inflammation) and phenotype (white vs. non-white). A sensitivity of 88.24% and a specificity of 100% were achieved utilizing Western blot analysis to detect M. a. avium infection in ring-neck doves, offering a negative predictive value of 93% and a positive predictive value of 100%. While Western blot analysis results did not reflect lesion severity, lesion type did partially correspond with the humoral response. The findings of the present study indicate that serologic testing can be used as a valuable ante mortem screening tool for identifying ring-neck doves infected with M. a. avium.