Isoenzyme variation in piroplasms isolated from bovine blood infected with Theileria annulata and T parva.

Blood from calves infected with Theileria annulata and T parva was freed from host cell elements and the piroplasms liberated from the red cells by ammonium chloride lysis. Lysates of the purified piroplasms and control host cell material were examined electrophoretically for several enzymes. Zymograms stained for glucose phosphate isomerase showed distinct differences between the host cell enzyme pattern and parasite enzyme patterns. The isoenzyme pattern of T annulata piroplasms differed from the isoenzyme pattern of T parva piroplasms.     

The clinical features and short-term treatment outcomes of scirrhous cord: A retrospective study of 32 cases

Scirrhous cord (SC) is an uncommon complication of castration, characterised by chronic infection of the spermatic cord remnant. It is reported that surgical excision of the infected tissue is the most effective means of treatment, but there are few published studies assessing the outcomes of horses treated for SC. The aims of this retrospective study were to describe the clinical features and short-term outcomes in horses treated for SC at two equine hospitals in the UK. The clinical records of horses diagnosed with SC over a 10-year period were reviewed. A diagnosis of SC was made if the gelding presented with typical clinical signs with confirmation at surgery. Thirty-two cases of SC were identified at the two equine hospitals. The mean age at presentation was 6 years (range 2–14 years, n = 22), and the median time from castration to presentation was 29.5 days (range 20–2500 days). Mean age at castration was 4.3 years (range 6 months to 10 years, n = 10). Clinical signs included scrotal swelling, discharging wounds, hindlimb lameness and pyrexia. Five horses demonstrated hyperfibrinogenaemia (n = 8). Microbial culture isolated various bacterial species. All 32 cases were treated with surgical excision of the infected tissue and discharged from the hospitals between 1 and 10 days post-operatively. A limitation of this study is that it was a retrospective study with no long-term follow-up available. It was concluded that the results of this study confirm that SC can present at variable time points following castration, even many years later, and that a variety of bacterial species may be involved. Surgical excision of infected tissue is a successful treatment with a good short-term prognosis for survival.     

Effect of intravenous infusion of a 7.2% hypertonic saline solution on serum electrolytes and osmotic pressure in healthy beagles.

The effect of an intravenous (i.v.) infusion of hypertonic saline solution (HSS; 7.2%, 2,400 mOsmol/kg.H2O) was evaluated by serum electrolyte concentrations and osmotic pressure in the anesthetized beagles. Sixteen beagles were assigned to 3 experimental groups (2.5, 5 or 15 ml/kg of HSS i.v. infusion) or a control group (5 ml/kg of isotonic saline solution (ISS) i.v. infusion) and were monitored for 120 min after the initiation of fluid infusion. The relative plasma volume (rPV) in the 5 ml/kg and 15 ml/kg HSS groups progressively expanded to 143.1 +/- 7.4% at 3 min and 156.4 +/- 5.9% at 5 min after the initiation of the fluid infusion, respectively. Significant increases were not produced by ISS and 2.5 ml/kg HSS infusion. The serum sodium and chloride concentrations in the ISS group were not altered. The 5 ml/kg HSS infusion induced transient high osmotic and sodium levels, and the serum sodium concentration remained under the 160 mM/l after the completion of the HSS infusion. However, the 15 ml/kg HSS infusion induced a constant high osmotic level (340.5-352.8 mOsmol/kg.H2O) and hypernatremia (161.4-174.5 mM/l) from 10 to 90 min after the initiation of the fluid infusion. The 15 ml/kg HSS infusion induced significant decreases in the partial pressure of oxygen (PaO2), reaching 63.7 +/- 8.0 mmHg at 120 min after the initiation of the fluid infusion compared with an immediately before fluid infusion value. On the basis of these findings, 5 ml/kg HSS infusion can be safely administered to healthy beagles for expanding the plasma volume without inducing hypernatremia. A 5 ml/kg HSS infusion is thus recommended for the initial field resuscitation of dogs.     

Histopathological and immunohistochemical analysis of the endocrine and exocrine pancreas in twelve cattle with insulin-dependent diabetes mellitus (IDDM).

Histological and immunohistochemical studies were carried out on the pancreas of twelve cattle of insulin-dependent diabetes mellitus (IDDM). They showed clinical signs such as persistent hyperglycemia, glycosuria and decreased glucose tolerance, and some cases accompanied with or without ketonuria. Histopathologically, eight cattle were diagnosed as chronic IDDM, while others were acute IDDM. The most characteristic lesions of the pancreas in chronic IDDM showed a decrease in the size and number of pancreatic islets, interlobular and interacinar fibrosis, mild lymphocytic insulitis, and vacuolation of a few islets. Almost all cells in the atrophied islets had a small amount of ungranulated cytoplasm. Immunohistochemical examination revealed that the atrophied islet cells did not react to anti-insulin antibody, but occasionally reacted to anti-glucagon or somatostatin antibodies. A few solitary islets with mild lymphocytic infiltration, necrotic islets with occasional calcification, and atrophied islets with mild fibrosis were also observed. A few islets consisted of many islet cells with vacuolated cytoplasm including a small number of insulin-positive granules. Accumulation of glycogen granules was occasionally observed in these islets. Islet fibrosis was due to the proliferation of collagen fibers reactive to both anti-collagen type I and type III antibodies. In acute IDDM, the major islets consisted of the cells with vacuolated cytoplasm indicating the degranulation of islet cells. These islets contained many islet cells with shrunken cytoplasm and karyorrhectic nuclei. Lymphocytic infiltration was frequently observed in the islets which consisted of many islet cells having karyorrhectic nuclei and vacuolated and severely degranulated cytoplasm. Immunohistochemically, islet cells with vacuolated cytoplasm had a small amount of insulin-positive granules, suggesting severe degranulation of beta-cells. An increase in acinar islet-cells and proliferation of ductal epithelial cells showing insulin-immunoreactivity were observed. Bovine IgG-immunoreactive islet cells were frequently seen in the vacuolated islets. In summary, pathological observations suggested that beta-cells were being destroyed by an inflammatory process which selectively affected the pancreatic islets. Lymphocytic insulitis and anti-bovine immunoreactive islet cells were thought to be the most significant changes in determining the etiology and pathogenesis of bovine IDDM, and suggested their role in anti-islet autoimmunity in this form of diabetes.     

Comparative pathology and pathogenesis of spontaneous and experimentally induced fibropapillomas of green turtles (Chelonia mydas)

Tumor biopsy samples from 25 Floridian and 15 Hawaiian green turtles (Chelonia mydas) with spontaneous green turtle fibropapillomatosis (GTFP) and from 27 captive-reared green turtles with experimentally induced GTFP were examined microscopically to differentiate the histologic features that result from GTFP pathogenesis and those that result from incidental factors that may vary according to geographic region. Common histologic features for spontaneous and experimentally induced tumors included fibroblast proliferation in the superficial dermis, epidermal acanthosis and hyperkeratosis, epidermal basal cell degeneration with dermal-epidermal cleft formation, spinous layer degeneration with intraepidermal vesicle and pustule formation, and ulceration. Visceral tumors, found in eight of 10 (80%) free-ranging turtles with cutaneous disease that were examined after death, had extensive interstitial fibrous proliferation. The presence of spirorchid trematode eggs and associated foreign body granulomas, common secondary findings within spontaneous tumors, varied by geographic location, and these findings were not observed in experimentally induced tumors. Eosinophilic intranuclear inclusions and intranuclear herpesvirus-associated antigen immunoreactivity were found in 18 of 38 (47%) experimentally induced cutaneous tumors and nine of 119 (7.5%) spontaneous tumors from Floridian but not Hawaiian turtles. The possible involvement of GTFP-associated herpesvirus in the pathogenesis of epidermal degenerative changes and GTFP pathogenesis is discussed.     

Effects of dietary level of ruminally protected choline on performance and carcass characteristics of finishing beef steers and on growth and serum metabolites in lambs

Ruminally protected choline (RPC) was evaluated in two experiments. In Exp. 1, beef steers (n = 160; average initial BW = 350.9 kg) were fed a 90% concentrate diet with either 0, .25, .5, or 1.0% RPC (DM basis) for 112 to 140 d. Feeding .25% RPC increased ADG 11.6% compared with 0% RPC, but responses diminished with increasing RPC level (cubic response, P < .10). Daily DMI was not affected by RPC level, but feed:gain was improved 6.8% with .25% RPC compared with 0% RPC, and responses diminished with increasing RPC level (cubic response, P < .10). Carcass yield grade increased linearly (P < .10) as RPC level increased, but marbling score was lower for all three RPC-containing diets than for the 0% RPC diet (quadratic response, P < .05). In Exp. 2, 20 Suffolk lambs (initial BW = 29.8 kg) were fed an 80% concentrate diet for 56 d with the same RPC levels as in Exp. 1. Serum triglycerides (TG) and cholesterol (CLSTRL) were measured in weekly blood samples, and intensive blood samples were collected on d 28 and 56 to evaluate serum insulin (INS), GH, and NEFA. For the 56-d feeding period, ADG responded quadratically (P < .10) to RPC level, but DMI and feed:gain were not affected. Serum INS and NEFA concentrations increased linearly (P < .05) and serum GH responded cubically (P < .05) to RPC level on d 28, but no differences were noted on d 56. Serum TG concentrations in weekly samples increased linearly (P < .10) with RPC level, but, averaged over all weeks, serum CLSTRL concentrations did not differ (P > .10) among treatments. Quantities of carcass mesenteric (P < .05) and kidney fat (P < .10) increased linearly, but longissimus muscle and liver fat contents did not differ (P > .10) among RPC levels. Supplementing RPC in high-concentrate diets improved performance, but results were not consistent among RPC levels or between cattle and sheep. Potential effects of RPC might be mediated through alterations in fat metabolism and(or) metabolic hormones related to fat metabolism.     

A Comparison of the Plasma Fructose Concentrations in Dogs and Cats and Changes in the Fructose Concentrations in Dogs Following Intravenous Administration of Fructose

The plasma concentrations of fructose, glucose, free fatty acids (FFA) and triglycerides (TG) were measured in dogs and cats. Changes in these concentrations were investigated in dogs by an intravenous fructose tolerance test (IVFTT) at a dose of 0.1 g/kg body weight. Fructose concentrations in the plasma of dogs were significantly higher than those of cats. There was no significant difference in plasma glucose concentrations between dogs and cats. Plasma FFA concentrations decreased and TG concentrations increased after feeding in both dogs and cats. During the IVFTT, the plasma fructose concentrations in the dogs increased rapidly to a peak by 2 min and then decreased to half of the peak by 5 min after the administration of fructose. Administration of fructose resulted in an increase in the plasma TG concentrations and reduced plasma FFA concentrations in the dogs. Only 4% of the administered fructose was detected in the urine of dogs following IVFTT. Plasma fructose was considered to be rapidly absorbed and metabolized in both dogs and cats. However, as with glucose metabolism, there appear to be some differences in fructose metabolism between dogs and cats.     

A serosurvey of Borna disease virus infection in wild rats by a capture ELISA

For a serological diagnostic test for Borna disease (BD), we developed a capture ELISA with specificity and sensitivity based on detection of antibodies against BD virus (BDV) p40 protein. Using our capture ELISA system, the antibody response of rats inoculated intracerebrally with BDV at 4 weeks after birth showed a sharp increase from 1 to 4 weeks postinoculation (p.i.) and a steady level after 5 weeks p.i. To investigate prevalence of BDV infection among wild rats, we examined sera of Rattus norvegicus in Kami-iso town, Oshima district, Hokkaido, suggesting that rats in this area had not been infected by BDV.     

Detection and identification of Mycoplasma conjunctivae in infectious keratoconjunctivitis by PCR based on the 16S rRNA gene.

A specific PCR assay based on unique sequences of the rrs genes (16S rRNA) of Mycoplasma conjunctivae was developed for direct detection and identification of this pathogen from clinical material. DNA from eye swabs was amplified after a simple lysis step by either a single PCR with the M. conjunctivae specific primer pair McoR1 and McoF1, or by a nested PCR with the Mycoplasma genus specific primer pair MOLIGEN1-L and 16UNI-R in the first step and McoR1 and McoF1 in the second step. The specificity of the primer pair McoR1 and McoF1 was verified with purified DNA from the type strain, from 17 field isolates of M. conjunctivae and from several Mollicutes which are phylogenetically related to M. conjunctivae or which can be isolated from the same host animals. This method identified mycoplasma isolates from goat, sheep, ibex and chamois originating from different countries as M. conjunctivae. No cross amplifications with other mycoplasmas which are related to M. conjunctivae were observed. Eye swab samples containing known numbers of M. conjunctivae cells were analysed after direct lysis of the material. The detection level was estimated to be 20 cells per swab when the nested PCR procedure was used and 2 x 10(5) by the single PCR method. In an experimental infection model of sheep, the nested PCR method for detection of M. conjunctivae gave results which were comparable to mycoplasmal culture. These are the implications for diagnostic purposes: M. conjunctivae isolates can be identified by the one-step PCR method, whereas for detection and identification of M. conjunctivae in clinical material the two-step method should be used (higher sensitivity).