Enzyme immunohistochemical staining of formalin-fixed tissues for diagnosis in veterinary pathology

Disease diagnosis often relies on the detection of specific antigens in tissue specimens. Enzyme-based immunohistochemical stains of formalin-fixed, paraffin-embedded tissues may be used to identify antigens associated with viral, bacterial and protozoal microorganisms, autoimmunity, and neoplasia. The detection of antigens in routinely fixed tissues offers several advantages over other diagnostic techniques. Sample submission is convenient and facilitates safe handling of potential human pathogens. Retrospective studies of stored specimens are possible. The technique is relatively rapid and enables detection of nonviable microorganisms. In addition, the ability to detect antigens in fixed specimens allows simultaneous visualization of the antigen and the histological lesion which may enhance the accuracy of diagnosis.     

Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome.

Postweaning multisystemic wasting syndrome (PMWS), an apparently new disease, has been recognized in swine herds in western Canada. Young pigs with this disease have progressive weight loss, tachypnea, dyspnea, and jaundice, accompanied by interstitial pneumonia, lymphadenopathy, hepatitis, and nephritis. We examined more than 400 pigs from more than 70 herds in Alberta, Saskatchewan, and Manitoba with cases of PMWS. A small virus was isolated from a range of tissues from 8 of 8 affected pigs examined. The agent was identified as a circovirus-like virus using electron microscopy, immunohistochemical staining with porcine and rabbit immune serum, and in situ hybridization. Immunohistochemical examination of tissues from more than 100 affected pigs has revealed widespread viral antigen, often contained in circovirus-like inclusion bodies, in lesions from numerous organs. Although Koch's postulates remain to be fulfilled, these results demonstrate a high degree of association between the presence of the circovirus-like virus and PMWS in affected swine.     

Campylobacter jejuni abortions in two beef cattle herds in Saskatchewan

Abortions, accompanied by placental retention and weight loss, occurred during February and March in 19% of 120 and 10% of 108 beef cows and heifers on two neighboring ranches in southern Saskatchewan. A diagnosis of Campylobacter jejuni abortion was made based on lesions of necrotizing and suppurative placentitis and fetal bronchopneumonia in association with the culture of large numbers of C. jejuni from placentas and fetal tissues.

Campylobacter jejuni was isolated with variable frequency from fecal samples of aborting and healthy cows, and scouring and healthy calves. Campylobacter jejuni serotype 2 (Lior) was isolated from fetal tissues and feces of a scouring calf, whereas C. jejuni serotypes 1, 4, 5 and 99 were isolated from feces of in-contact cattle. We hypothesized that the source and mode of transmission of C. jejuni was fecal contamination of water supplies and feeding grounds by carrier cows or wildlife.

     

Computed tomography diagnosis of eight dogs with brain infarction

Objective Medical records of eight dogs presenting with acute onset of neurological signs and a diagnosis of brain infarction as determined by computed tomography (CT) imaging were reviewed. Design Retrospective single-centre case review Results Ischaemic infarction in the territory of the rostral cerebellar artery was identified in three spaniel-breed dogs. All cerebellar infarcts were non-haemorrhagic. Telencephalic infarcts were identified in five dogs, in the territories of the middle cerebral artery (2/5) and rostral cerebral artery (3/5). One of these dogs had an ischaemic infarction, but all other infarctions appeared haemorrhagic. All dogs were geriatric (≥8 years old), with concurrent medical conditions identified in six dogs. One dog was euthanased after diagnosis because of the severity of its neurological signs and one dog was euthanased as a result of associated renal disease 2 months after diagnosis. Six dogs were alive at least 1 year after diagnosis. Conclusions CT is useful in the diagnosis of cerebrovascular accident in dogs, which can present as a spectrum of images with early changes in attenuation and subtle mass effects detected after infarction. CT is particularly sensitive for detecting haemorrhagic infarction, but under-represent ischaemic and lacunar infarctions when compared with MRI.     

Organochlorine residues in brook trout from remote lakes in the Northeastern United States

Organochlorine residues were analyzed in brook trout (Salvelinus fontinalis) collected from six undisturbed lakes in Maine, New Hampshire and Vermont. The residues (whole body) were compared among age groups of fish and were correlated by regression analyses with water chemistry factors that might be affected by acid precipitation. There were significant differences in the organochlorine content of fish from different lakes but not among age groups of fish within each lake or among all lakes. There were no significant correlations between the water chemistry values measured and organochlorine content of the fish. Inasmuch as no organochlorine pesticides have been used within the lake watersheds, the compounds must have reached these lakes by atmospheric transport.     

Evidence of fish population responses to acidification in the Eastern United States

The hypothesis that acidification has reduced or eliminated fish populations in certain areas of the eastern United States was investigated by examining present and historical fishery survey records. The causes of acidification (e.g., atmospheric deposition) were not specifically considered, although instances of obvious alternative explanations (e.g., acid mine drainage, organic acids) were avoided. The number of usable data sets located was small. Trend analyses are severely limited by the lack of high quality historical data. The strongest evidence for fisheries declines associated with acidification is provided by data for the Adirondack Mountain region of New York. In some lakes, fish populations have declined or disappeared; lakes experiencing fishery declines are now acidic. Alternative explanations for changes in fish communities over time were examined. In 49 lakes, some or all fish populations have apparently been lost with no available explanation other than acidification. Extrapolation of these data to the entire Adirondack Mountain region suggests that perhaps 200 to 400 lakes may have lost fish populations as a result of acidification. Streams in Pennsylvania and Massachusetts also had documented declines in fish populations that were associated with acidity; however, the data are fewer and less complete than those for New York. Acidification effects on fishery resources in other regions of the eastern United States are apparently minimal. The extent of the damage to date appears small relative to the total resource.     

The Effect of Fire on Mercury Cycling in the Soils of Forested Watersheds: Acadia National Park,Maine, U.S.A.

This study compares mercury (Hg) and methylmercury (MeHg) distribution in the soils of two forested stream watersheds at Acadia National Park, Maine, U.S.A. Cadillac Brook watershed, which burned in 1947, has thin soils and predominantly deciduous vegetation. It was compared to the unburned Hadlock Brook watershed, with thicker soil and predominantly coniferous vegetation. Soils in both watersheds were primarily well drained. The fire had a significant impact on the Cadillac watershed, by raising the soil pH, altering the vegetation, and reducing carbon and Hg pools. Total Hg content was significantly higher (P > 0.05) in Hadlock soils (0.18 kg Hg ha^-1) compared to Cadillac soils (0.13 kg Hg ha^-1). Hadlock O horizon had an average Hg concentration of 134±48 ng Hg g^-1 dry weight, compared to 103±23 ng Hg g^-1 dry weight in Cadillac O horizon. Soil pH was significantly higher in all soil horizons at Cadillac compared to Hadlock soils. This difference was especially significant in the O horizon, where Cadillac soils had an average pH of 3.41±0.22 compared to Hadlock soils with an average pH of 2.99±0.13.To study the mobilization potential of Hg in the O horizons of the two watersheds, batch adsorption experiments were conducted, and the results were modeled using surface complexation modeling. The results of Hg adsorption experiments indicated that the dissolved Hg concentration was controlled by the dissolved organic carbon (DOC) concentration. The adsorption isotherms suggest that Hg is more mobile in the O horizon of the unburned Hadlock watershed because of higher solubility of organic carbon resulting in higher DOC concentrations in that watershed.Methylmercury concentrations, however, were consistently higher in the burned Cadillac O horizon (0.20±0.13 ng Hg g^-1 dry weight) than in the unburned Hadlock O horizon (0.07±0.07 ng Hg g^-1 dry weight). Similarly, Cadillac soils possessed a higher MeHg content (0.30 g MeHg ha^-1) than Hadlock soils (0.16 g MeHg ha^-1). The higher MeHg concentrations in Cadillac soils may reflect generally faster rates of microbial metabolism due to more rapid nutrient cycling and higher soil pH in the deciduous forest. In this research, we have shown that the amount of MeHg is not a function of the total pool of Hg in the watershed. Indeed, MeHg was inversely proportional to total Hg, suggesting that landscape factors such as soil pH, vegetation type, or land use history (e.g., fire) may be the determining factors for susceptibility to high Hg in biota.     

Anthelmintic activity of paraherquamide in calves.

Paraherquamide, an oxindole alkaloid metabolite of Penicillium paraherquei and Penicillium charlesii, was tested against the adult stages of nine common gastrointestinal and lung nematodes of calves at single, oral dosages of 0.5, 1.0, 2.0 or 4.0 mg kg-1. At dosages 1.0-4.0 mg kg-1 there was 95% or more removal of Haemonchus placei, Ostertagia ostertagi, Trichostrongylus axei, Trichostrongylus colubriformis, Cooperia oncophora, Nematodirus helvetianus, Oesophagostomum radiatum, and Dictyocaulus viviparus. Cooperia punctata, the dosage-limiting species, was virtually unaffected by any dosage except the highest, which produced an efficacy of 89%. The 0.5 mg kg-1 dosage was 95% or more efficacious against H. placei, O. ostertagi, C. oncophora, and D. viviparus, but weaknesses were evident against the other five species. No adverse reaction was observe in any calf.     

Detection of Mycobacterium paratuberculosis in formalin-fixed paraffin-embedded tissues by the polymerase chain reaction.

The polymerase chain reaction (PCR) utilizing primers specific for the IS900 sequence of Mycobacterium paratuberculosis was applied to tissue sections of formalin-fixed, paraffin-embedded ileum from cattle with Johne's disease and the results compared to those obtained with acid-fast (Ziehl-Neelsen) and immunohistochemical staining. The PCR was positive in 19/21 tissues (90%) while Ziehl-Neelsen staining was positive in 18/21 (86%) and immunohistochemical staining in 21/21 (100%). The Ziehl-Neelsen and immunohistochemical stains are not, however, specific for M. paratuberculosis. The PCR for detection of M. paratuberculosis using the IS900 sequence is a specific and relatively sensitive method for confirmation of Johne's disease and its application to formalin-fixed, paraffin-embedded tissues may be useful for confirmation of dubious cases, for retrospective studies and for epidemiological analyses.