DNA profiling reveals remarkably low genetic variability in a herd of SLA homozygous pigs.

Inbred strains of rodents have become indispensable for a wide range of biological studies. It has generally been accepted that genetic uniformity is unlikely to be achieved before 20 generations of brother x sister matings discouraging attempts to inbreed larger mammals. Nevertheless, pigs, homozygous for the swine MHC haplotype SLA b/b, have been inbred at the Babraham Institute for almost thirty years and used for immunological studies. Since the herd had not been studied at the DNA level, DNA profiling at multiple hypervariable loci was performed and surprisingly little genetic polymorphism and extremely high inter-individual resemblance were observed reminiscent of that observed in inbred strains of mice.     

Comparison of the supplemental effects of Saccharomyces cerevisiae and antibiotics in low-protein and high-fibre diets fed to broiler chickens.

Two experiments were completed to compare the supplemental effects of yeast Saccharomyces cerevisiae and sub-therapeutic antibiotics in high-fibre and low-protein diets for broiler chicks. In experiment 1, yeast was added at 1.5, 3.0 and 6.0 g kg-1 while penicillin, tylosin or neoterramycin were added at 150 mg kg-1 into different batches of a high-fibre diet containing 250 g kg-1 palm kernel meal. Saccharomyces cerevisiae and antibiotics increased BW, BWG, FCR, feed intake, carcass and breast weights above the control. Small intestine weight was reduced by antibiotics, while yeast caused a lower deposition of abdominal fat. Liver weights of different treated broilers were similar. The performance indices were superior on penicillin and 3.0 g kg-1 yeast compared to other supplements evaluated. In the second experiment, 1.5 and 3.0 g kg-1 of yeast, and 0.75 mg kg-1 of procaine penicillin, zinc bacitracin and tylosin were separately added to a 180 g kg-1 crude protein diet. Performance of the chicks with additives was compared with unsupplemented negative and positive controls containing 180 and 210 g kg-1 crude protein respectively. Broilers fed with supplements had superior BW, BWG, FCR and feed intake compared to the negative control, but carcass data were similar. Dietary penicillin increased BW, BWG, feed intake more than other supplements, but yeast stimulated higher FCR. Data from both studies indicated that Saccharomyces cerevisiae comparatively increased growth and carcass weight and reduce abdominal fat, therefore, it can serve as a natural substitute for antibiotics in broiler diet.     

Feline retinal degeneration: clinical experience and new findings (1994-1997)

A retrospective case series of 26 cats with diffuse retinal degeneration is presented. The most common presenting complaints included bumping into objects, dilated pupils, and reluctance to jump. Ophthalmic examination findings were consistent with those reported in dogs with progressive retinal atrophy. Breed predilection of the Siamese cat was observed. Cats with primary retinal degeneration presented late in the clinical course of their disease, when vision loss was severe. Early symptoms such as night blindness and secondary ocular complications (i.e., cataract and retinal detachment), reported in dogs with progressive retinal degeneration, were not observed in this study. All cats showed excellent adaptive capabilities to blindness.     

The detection of CD2+, CD4+, CD8+, and WC1+ T lymphocytes, B cells and macrophages in fixed and paraffin embedded bovine tissue using a range of antigen recovery and signal amplification techniques

In order to develop procedures to label the main bovine leucocyte populations in paraffin embedded sections, the immunoreactivity of 25 monoclonal antibodies (mAbs) to different leucocyte antigens was assessed with formal dichromate (FD5) and 10% formalin fixation, a battery of antigen retrieval (AR) methods, and the biotin-tyramide amplification system. All the leucocyte populations investigated (CD2+, CD4+, CD8+, WC1+ T lymphocytes, B cells and macrophages) were strongly and specifically detectable under an appropriate combination of mAb, AR method and signal amplification system. CD4 and CD8 required the most stringent conditions and could only be demonstrated in FD5 fixed sections. For detection of CD2, WC1+ T lymphocytes, B cells and macrophages, all the mAbs produced immunoreactivity in FD5 or formalin fixed tissues. The need to check a range of different AR methods is stressed, as the method of choice varied for each individual mAb. The incorporation of the signal amplification system was necessary to observe a strong signal and the complete distribution of CD4, CD8 and B cells. Fixation by FD5 proved to be better than formalin for the preservation of surface antigens but it was inferior for the detection of markers which were found to show cytoplasmic immunoreactivity, such as the macrophage marker MAC387 or the B cell markers BAQ155 or IL-A59.     

Molecular heterogeneity in clinical isolates of Malassezia pachydermatis from dogs

Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far.     

REVERSIBLE MAGNETIC RESONANCE IMAGING ABNORMALITIES IN DOGS FOLLOWING SEIZURES

Reversible magnetic resonance (MR) imaging lesions have been described in humans following seizures. This condition has not yet been reported in animals. This paper describes reversible abnormalities identified in 3 dogs using MR imaging that was performed initially within 14 days of the last seizure and follow-up imaging that was performed after 10 to 16 weeks of anticonvulsant therapy. All three dogs had lesions in the piriform/temporal lobes, characterized by varying degrees of hyperintensity on T2-weighted images and hypointensity on T1-weighted images. In one dog, contrast enhancement was evident. On reevaluation, partial resolution occurred in all 3 dogs. In a fourth animal with an olfactory meningioma, similar appearing lesions in the temporal cortex and right and left piriform lobes were identified after seizure activity. A surgical biopsy of the temporal cortex and hippocampus was performed and edema, neovascularization, reactive astrocytosis, and acute neuronal necrosis were evident. These histologic findings are similar to those reported in humans with seizures. Recognizing the potential occurrence of reversible abnormalities in MR images is important in developing a diagnostic and therapeutic plan in canine patients with seizures. Repeat imaging after seizure control may help differentiate between seizure-induced changes and primary multifocal parenchymal abnormalities.     

Feeding-induced increases in insulin do not suppress secretion of growth hormone

Secretion of growth hormone (GH) is reduced for several hours after feeding when access to feed is restricted to a 2-hr period each day. We hypothesized that increased secretion of insulin after feeding inhibits release of GH from the anterior pituitary gland. Our objectives were to determine whether: 1) alloxan prevents concentrations of insulin from increasing after feeding steers; 2) concentrations of GH remain high after feeding alloxan-treated steers; and 3) GH-releasing hormone (GHRH) stimulates greater release of GH in alloxan-treated, than in control, steers after feeding. Steers were injected iv with either saline (control) or with alloxan (110 mg/kg) (n = 4 per group). Concentrations of insulin were not different (P = 0.61) between control and alloxan-treated steers before feeding (87.5 +/- 33.6 pmol/l). However, alloxan prevented insulin from increasing (P < 0.001) after feeding (131.8 pmol/1) compared with control steers (442.0 pmol/l) (pooled SEM = 47.5). Overall, GH was higher (P < 0.05) in alloxan-treated (6.4 ng/ml) than in control steers (3.7 ng/ml) (pooled SEM = 0.7), but GH decreased (P < 0.001) after feeding in both groups. Iv injection of GHRH stimulated release of GH 1 hr before, but not when injected 1 hr after feeding (P < 0.001). In addition, net areas under the GH curve were not significantly different between control and alloxan-treated groups. We conclude that increased concentrations of insulin after feeding do not mediate feeding-induced suppression of GH secretion in steers.     

Comparison of clinical signs and hemodynamic variables used to monitor rabbits during halothane- and isoflurane-induced anesthesia

OBJECTIVE: To characterize variables used to monitor rabbits during inhalation anesthesia. ANIMALS: 8 male New Zealand White rabbits. PROCEDURE: Rabbits were similarly anesthetized with halothane (HAL) or isoflurane (ISO) in a crossover study; half received HAL followed by ISO, and the protocol was reversed for the remaining rabbits. After induction, minimum alveolar concentration (MAC) was determined for each agent, using the tail-clamp method, and variables were recorded at 0.8, 1.0, 1.5, and 2.0 MAC (order randomized). RESULTS: Mean +/- SEM MAC was 1.42 +/- 0.05 and 2.07 +/- 0.09% for HAL and ISO, respectively. Directly measured auricular mean arterial blood pressure was 52.8 +/- 5.6 and 54.8 +/- 6.1 mm Hg at 0.8 MAC for HAL and ISO, respectively, and decreased from these values in a parallel dose-dependent manner. Respiratory frequency remained constant (range, 69 to 78 breaths/min) over the range of HAL doses but incrementally decreased from a mean of 53 (at 0.8 MAC) to 32 breaths/min (at 2.0 MAC) for ISO. The PaCO2 was similar at 0.8 MAC for HAL and ISO and progressively increased with increasing doses of both agents; PaCO2 at 2.0 MAC for ISO was significantly greater than that at 2.0 MAC for HAL (79.8 +/- 13.7 vs 54.9 +/- 4.0 mm Hg, respectively). Eyelid aperture consistently increased in a dose-dependent manner for both anesthetics. CONCLUSIONS: Arterial blood pressure, PaCO2, and eyelid aperture consistently and predictably changed in rabbits in response to changes in anesthetic doses. The magnitude of respiratory depression was greater for ISO than for HAL.     

The effects of prednisone and azathioprine on circulating immunoglobulin levels and lymphocyte subpopulations in normal dogs.

This study investigates serum immunoglobulin (SIg) levels and lymphocyte subpopulations in normal dogs in response to putative immunosuppressive doses of prednisone and/or azathioprine. The objectives were to quantify SIg levels and lymphocyte subpopulations, including Thy-1+, CD4+, CD8+ and B cells, in normal dogs both before and after the administration of prednisone and/or azathioprine at 2 mg/kg, PO, each. Eighteen beagles were divided into 3 groups of 6 dogs each. Blood samples for radial immunodiffusion assay of IgG, IgM and IgA, complete blood count (CBC)and flow cytometry were collected prior to the administration of any drugs and again after 14 d of azathioprine, prednisone or azathioprine and prednisone. Peripheral blood mononuclear cells were isolated using density centrifugation and were incubated with monoclonal antibodies reacting with CD4+, CD8+, Thy-1+ and membrane immunoglobulin. Lymphocyte subsets were quantified using flow cytometry. Azathioprine-treated dogs had no significant changes in SIg levels or lymphocyte subpopulations. Prednisone-treated dogs had significant (P < 0.05) decreases in all SIg levels, all lymphocyte subpopulations and erythrocyte numbers, and had an increase in neutrophil counts. Prednisone and azathioprine-treated dogs had significant (P < 0.05) decreases in serum IgG levels and Thy-1+ and CD8+ lymphocyte subpopulations, with an increase in the CD4:CD8. These dogs also had a significant decrease in erythrocyte number and a significant increase in the monocyte count. These findings suggest that azathioprine and prednisone in combination or prednisone alone may be useful for the treatment of T cell-mediated diseases since decreased circulating T cell levels were demonstrated following treatment. The combination of drugs or azathioprine alone may not be appropriate for treatment of acute or autoantibody-mediated immune disease, because SIg levels were minimally affected by treatment.     

The use of chance-corrected agreement to diagnose canine compulsive disorder: an approach to behavioral diagnosis in the absence of a 'gold standard'.

This study assessed the diagnostic accuracy of formal diagnostic criteria for canine compulsive disorder (canine CD). Canine CD is a syndrome of abnormal behaviors that are believed to result from conflict or frustration. Differential diagnoses include normal conflict behavior and learned behavior. In studies of canine CD, confidence in the diagnosis comes with knowing the accuracy of the diagnostic method. This accuracy may be quantified as the chance-corrected agreement between the diagnostic method and a 'gold standard' diagnostic test. The present study examined the agreement between diagnoses of canine CD made by an expert (the 'gold standard') and by using formal diagnostic criteria. The owners of 84 dogs suspected of having CD received 2 telephone interviews. The first utilized a detailed, pre-tested questionnaire; a dog was then diagnosed with CD if the behavioral history met 7 diagnostic criteria. The second interview was given by a behavioral expert whose diagnosis was based on personal experience. The interviewers were blind to each other's diagnoses. The chance-corrected agreement between diagnoses was minimal (kappa = 0.02) and disagreement was associated with 3 of the formal criteria: a history of conflict or frustration, an increase in the number of contexts that elicit the behavior, and an increase in the daily frequency of the behavior. Reasons for the disagreement include the order of the interviews, response biases, the setting of the interviews, and, possibly, the diversity of the behaviors associated with canine CD. To the authors' knowledge, this type of study is the first in clinical ethology to address validation of the diagnostic method. The results indicate 3 developmental aspects of canine CD that should be examined in future work.