Protein Electrophoresis of Serum and Heparinized Plasma in the Common Mynah (Acridotheres tristis).

Although serum protein electrophoresis is a diagnostic tool available through many veterinary laboratories, there currently are no reference intervals for protein fractions in healthy common mynahs (Acridotheres tristis). Therefore, electrophoretic patterns of proteins in serum and heparinized plasma of the common mynah were evaluated. Blood specimens were collected from 55 healthy adult common mynahs of unknown age (26 males and 29 females). The serum total protein and protein fractions were measured using the biuret method followed by cellulose acetate electrophoresis (CAE). The serum level of albumin was compared with bromocresol green (BCG) dye-binding and CAE methods. Four protein fractions, including albumin and α, ß, and γ globulins, were recorded in the electrophoretogram of serum specimens. Sex appeared to have no significant effect on the measured parameters. The serum BCG albumin fraction was significantly higher than the CAE albumin fraction (P = .01). Also, the comparison of total protein and protein fractions in serum and plasma specimens of 25 of the 55 birds sampled showed that total protein (Cohen index d = 0.66, P = .03), gamma globulin (d = 1.13, P = .00), and total globulin (d = 0.67, P = .00) in plasma samples were significantly higher than those in serum samples. The results of this study provide the specific reference intervals for total protein and protein fractions in common mynahs, which are essential for proper interpretation of laboratory results and also revealed that the albumin measurement by the BCG method yields unreliable results in common mynahs.

Evaluation of usefulness of a commercial agarose gel electrophoresis kit (QuickGel SP) for bovine serum protein electrophoresis.

The aim of this study was to show the usefulness of a commercial agarose gel electrophoresis (AGE) kit (QuickGel SP) for separating bovine serum protein fractions in comparison with conventional cellulose acetate electrophoresis (CAE). Serum protein bands were verified using five reference reagents corresponding to albumin and α1-, ß1-, ß2-, and γ-globulins. AGE clearly revealed six separated fractions of albumin and α1-, α2-, ß1-, ß2-, and γ-globulin fractions in 100% and 77.8% in serum samples of dairy cows from the healthy (n=27) and diseased groups (n=27), respectively. The α1- and α2-globulins were not separated by CAE in 14.8% and 96.3% of the samples from the healthy and diseased groups, respectively, whereas ß2- and γ-globulin were not separated by CAE in 96.3% and 100% of the samples from the healthy and diseased groups, respectively. More than 94% of the points for the α-globulin fractions (α1- and α2-globulins), the ß-γ-globulin fractions (ß1-, ß2-, and γ-globulins), and the albumin/globulin ratio between AGE and CAE were within agreement on the Bland-Altman plots. However, the mean biases were not near zero in the albumin and ß-γ-globulin fractions. These results suggest that the high-resolution commercial AGE kit can be utilized to separate bovine serum protein fractions.

Effect of storage time and temperature on the total protein concentration and electrophoretic fractions in equine serum.

Serum protein electrophoresis (SPE) is a technique that could be considered one of the most useful diagnostic aids available to the clinician. The effect of storage time and temperature on the total proteins and electrophoretic fractions (albumin, α1-, α2-, ß1-, ß2-, and γ-globulins) was assessed in 24 healthy horses. All samples, collected by jugular vein puncture, were centrifuged and divided into 4 aliquots. The 1st aliquot was analyzed within 3 h from collection (time 0), the 2nd was refrigerated at +4°C for 24 h, the 3rd was refrigerated at +4°C for 48 h, and the last was frozen at -20°C for 48 h. One-way repeated-measures analysis of variance (ANOVA) showed a significant effect (P < 0.05) of the different storage conditions on the concentrations of all the parameters studied and significant variations in the percentages of albumin, α1-globulins, α2-globulins, and γ-globulins. Compared with time 0 the total protein concentration increased significantly after 48 h at -20°C, the albumin percentage decreased after 48 h at -20°C, the α1-globulin percentage increased after 24 h at +4°C, the α2-globulin percentage increased after 48 h at +4°C and at -20°C, and the γ-globulin percentage increased after 48 h at -20°C. The results should help veterinary practitioners handle and store equine serum samples appropriately. Further investigations at different storage times and temperatures could be useful.

L'électrophorèse des protéines sériques (SPE) est une technique qui pourrait être considérée comme un des outils diagnostiques les plus utiles au clinicien. L'effet du temps et de la température d'entreposage sur les protéines totales et les fractions électrophorétiques (albumine, α1-, α2-, ß1-, ß2-, et γ-globulines) a été évalué chez 24 chevaux en santé. Tous les échantillons ont été prélevés par ponction de la veine jugulaire, centrifugés et divisés en quatre aliquots. Le premier aliquot a été analysé en dedans de trois heures du moment de la collecte (temps 0), le deuxième a été réfrigéré à 4 °C pour 24 h, le troisième a été réfrigéré à 4 °C pour 48, et le dernier a été congelé à −20 °C pendant 48 h. Une analyse de variance unidirectionnelle sur des mesures répétées (ANOVA) a montré un effet significatif (P < 0,05) des différentes conditions d'entreposage sur les concentrations de tous les paramètres étudiés et des variations significatives dans les pourcentages d'albumine, d'α1-globuline, d'α2-globuline, et de γ-globuline. Comparativement au temps 0, la concentration de protéines totales a augmenté significativement après 48 h à −20 °C, le pourcentage d'albumine a diminué après 48 h à −20 °C, le pourcentage d'α1-globuline a augmenté après 24 h à 4 °C, le pourcentage d'a2-globuline a augmenté après 48 h à 4 °C et à −20 °C, et le pourcentage de γ-globuline a augmenté après 48 h à −20 °C. Ces résultats devraient aider les vétérinaires praticiens à manipuler et entreposer de manière appropriée les échantillons de sérum équin. Des études ultérieures sur différents temps et températures d'entreposage seraient utiles.(Traduit par Docteur Serge Messier).

Diagnostic accuracy of electrophoretic analysis of native or defribrinated plasma using serum as a reference sample.

BACKGROUND: Electrophoretic analysis of plasma may provide inaccurate results unless plasma is defibrinated with ethanol. OBJECTIVE: The aim of this study, conducted following the Standards for Reporting of Diagnostic Accuracy (STARD), was to determine whether cellulose acetate (CAE), agarose gel (AGE), and capillary zone (CZE) electrophoresis of native plasma (NP) and defibrinated plasma (DP) provided the same information as serum electrophoretograms. METHODS: Serum, NP, and DP electrophoretograms obtained using the 3 methods were examined visually by 3 observers to identify changes in globulin fractions and provide diagnostic interpretations. Using serum analysis as the reference method, sensitivity, specificity, and positive likelihood ratio of NP and DP electrophoretograms to identify abnormal globulin profiles were calculated. RESULTS: Specimens from 46 dogs were analyzed. Albumin and α(1) - and γ-globulin fractions were lower and ß(2) -globulin fractions higher in NP than in serum and DP. Interpretations of NP electrophoretograms were the same as for serum in 54.8% (CAE), 51.2% (AGE), and 51.6% (CZE), revealed an increased ß-globulin fraction in 32.3-41.5%, and resulted in misinterpretation, especially for CZE analysis, in 4.7-16.1% of the dogs; all dogs with abnormal serum electrophoretograms were identified correctly using NP. Analysis of DP was similar to serum analysis in about 2/3 of the dogs. In the others, defibrination did not resolve spurious plasma findings or induced additional changes. Up to 75 and 27% of NP and DP electrophoretograms, respectively, were abnormal in dogs with normal serum electrophoretograms. Sensitivity for NP and DP analysis was high, but specificity for NP was poor. CONCLUSIONS: Analysis of NP provides the same information as serum analysis in > 50% of cases, and, despite low specificity, could preliminarily exclude the presence of abnormalities when only plasma is available. If electrophoretograms are not normalized by defibrination, electrophoretic abnormalities are likely present, and electrophoresis should be repeated using serum.

Electrophoretic serum protein fraction profile during the different physiological phases in Comisana ewes.

The aim of this study was to evaluate the influence of different physiological phases on serum total proteins and their fractions of ten Comisana ewes housed in Mediterranean area. From each animal, blood samples were collected at different physiological phases: late pregnancy, post-partum, early, mid-, end lactation and dry period. On all samples serum total proteins were determined by the biuret method, and albumin, α-globulins, ß(1) -globulins, ß(2) -globulins and γ-globulins concentrations were assessed using an automated system. One-way repeated measures analysis of variance was applied to determine the significant effect of different physiological phases on the parameters studied. During the late pregnancy and post-partum, total proteins, ß1- and ß2-globulins and γ-globulins showed the highest values. Starting from post-partum, α-globulins increased to reach their peaks in mid-lactation. Early lactation was characterized by low γ-globulins values. The increase in serum albumin concentration and the drop in some globulin fractions determined the significant increase in albumin/globulin ratio. The obtained results contributed to improve the knowledge on electrophoretic profile during the different physiological phases in ewes, confirming that pregnancy and lactation periods affect the protein metabolism. Particularly, serum protein fractions pattern could give information about dehydration, plasma volume expansion and hepatic function, which occur during the different physiological phases. Dynamics of the protein profile - from pregnancy to dry period - which are provided by our results, could be considered as guidelines for the management strategies to guarantee the nutritional needs of these animals during the different physiological phases and to avoid a decline of productive performance and consequently an economic loss.

Interpretation of capillary zone electrophoresis compared with cellulose acetate and agarose gel electrophoresis: reference intervals and diagnostic efficiency in dogs and cats.

BACKGROUND: Serum protein electrophoresis is widely used for diagnostic and research purposes. Cellulose acetate (CAE) and agarose gel (AGE) electrophoresis are the most frequently used methods, but capillary zone electrophoresis (CZE) is beginning to be used more in veterinary laboratories. However, reference intervals for CZE in animals and comparison studies with the other electrophoretic techniques are lacking, compromising the diagnostic utility of CZE. OBJECTIVES: The aims of this study were to compare results obtained using CAE, AGE, and CZE; to establish reference intervals for CZE in dogs and cats; and to assess the capacity of CZE to detect abnormalities identified by AGE. METHODS: Serum samples from 204 dogs, including 104 healthy animals, and 62 cats, including 28 healthy animals, were analyzed using automated systems for CAE, AGE, and CZE. Descriptive statistics and Passing-Bablok and Bland-Altman tests were used to compare results. For each technique, reference intervals were calculated based on results from healthy animals. Concordance between CZE and AGE in detecting pathologic changes was assessed using Cohen's k coefficient. RESULTS: For most protein fractions, values obtained by CAE, AGE, and CZE were significantly different from each other, and constant and proportional errors were often detected. Nevertheless, reference intervals obtained by the 3 techniques overlapped. Moreover, Cohen's k coefficient demonstrated that the capacity of CZE and AGE to detect pathologic changes was comparable. CONCLUSIONS: CZE performs comparably to AGE and CAE as long as CZE-specific reference intervals are used for interpretation and distinctive visual patterns for albumin, gaps between fractions, and subpeaks found on CZE tracings are recognized. In addition, CZE offers several technical advantages, such as ease of use and complete automation.

Detection of biclonal gammopathy by capillary zone electrophoresis in a cat and a dog with plasma cell neoplasia.

Gammopathies associated with plasma cell neoplasms in a 15-year-old female spayed domestic shorthaired cat and a 9-year-old female spayed Rottweiler dog were evaluated by serum protein electrophoresis. In the cat, the plasma cell neoplasm was found in the liver and spleen, and an evaluable sample of bone marrow was not obtained. Some of the plasma cells had the morphologic appearance of flame cells. The paraprotein was confirmed as IgG based on agar gel immunodiffusion precipitation and both immunocytochemical and immunohistochemical staining. The dog had multiple myeloma with production of IgG and IgA paraproteins. In both cases, serum proteins were evaluated by 2 methods of protein electrophoresis: cellulose acetate electrophoresis (CAE) and capillary zone electrophoresis (CZE). In the cat and the dog, CAE showed a single large oligoclonal-like peak, which occurred in the γ-region in the cat and the ß-γ-region in the dog, whereas CZE showed a biclonal gammopathy with 2 very close narrow spikes in the γ- and ß-γ-regions in the cat and dog, respectively. In selected cases, CZE may be more effective than routine CAE in distinguishing oligoclonal from monoclonal or biclonal paraproteinemia.

Plasma protein electrophoresis of Trachemys scripta and Iguana iguana.

BACKGROUND: Protein electrophoresis is widely applied in veterinary medicine, but is not used often in reptiles, in part because of lack of reference values. OBJECTIVE: The goals of this study were to compare plasma protein profiles obtained by cellulose acetate electrophoresis (CAE) and agarose gel electrophoresis (AGE), measure precision and examine interference by sample hemolysis, and establish preliminary reference intervals for 2 reptile species. METHODS: Heparinized plasma samples from healthy and diseased adult female Iguana iguana (n=40) and Trachemys scripta (n=60) were analyzed by CAE and AGE. Total protein concentration was measured by the biuret method. Electrophoresis results were compared using Bland-Altman plots and Passing-Bablok regression analysis. Precision and the effects of sample hemolysis were determined. Results from clinically healthy animals were used to determine reference intervals. RESULTS: Five protein fractions were identified in both species, with bisalbuminemia observed in 23/40 iguanas. High correlation was observed between the 2 methods for all fractions, with few proportional and systematic errors. Coefficients of variation were lower using AGE vs CAE and for I. iguana vs T. scripta. Two additional bands were observed in hemolyzed samples from T. scripta; 1 additional band was observed for I. iguana. Minimum and maximum values were reported for healthy I. iguana (n=14) and T. scripta (n=22). CONCLUSIONS: Although both methods are acceptable, the performance of AGE was slightly better than that of CAE for analysis of plasma from reptiles. Furthermore, reptile electrophoretic patterns should be interpreted based on the method used, the species analyzed, and the quality of the plasma sample.

Cellulose acetate electrophoresis of canine plasma after fibrinogen precipitation by ethanol.

BACKGROUND: In routine canine medicine, anticoagulated blood is often the only sample sent to laboratories for diagnostic purposes. This hampers the interpretation of protein electrophoretic tracings because plasma contains fibrinogen, which migrates in the beta-gamma region. In human medicine, fibrinogen can be precipitated from plasma using ethanol. OBJECTIVES: The purpose of this study was to assess ethanol precipitation as a method for removing fibrinogen from canine plasma so as to facilitate the interpretation of electorphoresis results. METHODS: Blood samples collected from 40 dogs were divided into plain tubes and tubes containing EDTA (n=20) or lithium-heparin (n=20). An aliquot of plasma from each sample was incubated with ethanol at a final concentration of 100 mL/L. Cellulose acetate electrophoresis was then performed on serum, plasma, and plasma treated with ethanol. To verify the efficiency of ethanol treatment, fibrinogen was added to 5 canine serum samples at final concentrations of 2.5, 5.0, and 10.0 g/L, and electrophoresis was performed before and after ethanol treatment. RESULTS: Visual analysis of electrophoretograms from ethanol-treated samples confirmed the disappearance of the fibrinogen peak from the beta(2)-globulin region. Treatment with ethanol caused a significant decrease in the percentage of beta(2)-globulins and a significant increase in the percentage of alpha(2)-globulins. Absolute values of most electrophoretic fractions were significantly decreased in ethanol-treated plasma compared with serum. CONCLUSIONS: Ethanol treatment successfully removed fibrinogen from canine plasma and normalized electrophoretic profiles, but probably also precipitated proteins other than fibrinogen. Ethanol treatment is recommended to facilitate visual identification of abnormal monoclonal peaks, but not for determining absolute protein concentrations in electrophoretic fractions.

Assessment of serum protein fractions in rainbow trout using automated electrophoresis and densitometry.

BACKGROUND: Although rainbow trout (Oncorhynchus mykiss, Walbaum) are one of the most-studied fish, electrophoretic techniques and classification of serum protein fractions have not been standardized, such that clinically useful values are lacking. OBJECTIVE: The aim of the present study was to evaluate preliminarily the serum protein fractions of rainbow trout using automated cellulose acetate electrophoresis and densitometry. METHODS: Serum samples from 25 rainbow trout (Oncorhynchus mykiss, Walbaum) were electrophoresed on cellulose acetate plates and quantified using densitometry. RESULTS: A maximum of 6 fractions were identified and numbered, in order of decreasing mobility, as I, II, III, IV, V, and VI. In 3 of 25 (12%) samples, 6 fractions were identified; in 18 (72%) samples, 5 fractions were identified; and in 4 (16%) samples, 4 fractions were identified. Fractions I, V, and VI were always clearly identifiable, whereas fractions II and IV were frequently fused and indistinguishable from fraction III. The pattern with 5 fractions was the most probable type (chi(2), P<.01). The mean (+/-SEM) protein concentrations of the 6 fractions were I, 0.8+/-0.1 g/dL; II, 0.3+/-0.0 g/dL; III, 1.6+/-0.1 g/dL; IV, 0.3+/-0.1 g/dL; V, 0.6+/-0.0 g/dL; and VI, 0.2+/-0.0 g/dL. Based on comparison of serum and plasma electrophoretic patterns from 8 fish, fibrinogen was found in fraction V. CONCLUSION: Automated cellulose acetate electrophoresis and densitometry appear to be a practical method for estimation of serum protein fractions in rainbow trout.

On the clinical chemistry of the Bennett's wallaby (Macropus rufogriseus rufogriseus).

The following parameters were determined in blood serum of apparently healthy Bennett's wallabies (Macropus rufogriseus) using the Hitachi 917 (Roche Diagnostics, Mannheim, Germany) and/or the Vettest 8008 (IDEXX-GmbH, Woerrstadt, Germany): alkaline phosphatase, alanine aminotransferase, ammonia, alpha-amylase, aspartate aminotransferase, Ca, Cl, cholesterol, cholinesterase, creatine kinase, creatinine, gammaglutamyltransferase, glucose, iron, lactate dehydrogenase, magnesium, phosphate, potassium, protein, sodium, total bilirubin, triglyceride, and urea. The results for cholesterol, glucose, total protein, triglyceride and for the enzymes alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, gamma-glutamyltransferase and lactate dehydrogenase differed significantly between both methods (P < 0.05). There is a negative correlation between the age of the Bennett's wallabies and the activity of the alkaline phosphatase. Five protein fractions could be separated on cellulose acetate electrophoresis. The mean concentrations of fructosamine and beta-hydroxybutyrate were 447.3 micromol/L and 0.27 mmol/L, respectively. The estimated vitamin A intake had no influence on the vitamin A concentration in serum. The serum vitamin E concentration was in general low and vitamin E was below the detection limit of 0.82 micromol/L in 29 out of 42 serum samples. The use of these analytes is discussed concerning the knowledge about the physiology, nutrition and diseases of macropods.

Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.

Galactosaminoglycan composition in chicken eggshell.

Galactosaminoglycans, isolated from decalcified chicken eggshell by papain digestion and ion-exchange chromatography, were fractionated by selective precipitation at varying concentrations of ethanol and characterized by chemical and enzymatic methods. The eggshell contained 0.15 microg galactosaminoglycan uronic acid/mg dry weight. Most (to approximately 87% of total) galactosaminoglycans were found to be chondroitin sulfate-dermatan sulfate copolymers with iduronic acid contents being approximately 20 to 30% of uronic acid. The remaining (to approximately 12% of total) galactosaminoglycans were chondroitin sulfate-dermatan sulfate copolymers with higher iduronic acid contents averaging 59% of uronic acid. Results of chondroitinase-ABC digestion demonstrated 4-sulfated disaccharides to be the major repeating units in the chicken eggshell galactosaminoglycans.

Trypanosomatidae from wild mammals in the neotropical rainforest of French Guiana.

The initial filling of the reservoir behind the Petit Saut hydro-electric dam, on the Sinnamary River in French Guiana, threatened the terrestrial and arboreal animals living in the neotropical rainforest being flooded. During a rescue programme between 24 October and 12 November in 1994, many of these animals were checked for infection with trypanosomatids. Overall, 45 blood samples and 54 skin biopsies were collected from 53 mammals (of 13 species representing five orders) and blood samples were also taken from each of nine reptiles (six species from four families). When the skin biopsies and the buffy-coats from the blood samples were cultured in NNN medium, 10 of the cultures, each initiated with mammalian blood, were found to be positive for trypanosomatids. Multilocus enzyme electrophoresis (MLEE) on cellulose acetate plates, with 20 enzyme systems, was then used to investigate each of the positive cultures. The results were analysed by clustering from a genetic distance matrix, using the unweighted pair group method with arithmetic averages (UPGMA), and applying a bootstrap procedure to Wagner parsimony trees. A stock obtained from Didelphis marsupialis was identified as a zymodeme of Trypanosoma cruzi (Miles' zymodeme 1) known to cause Chagas disease in French Guiana. Five stocks (one each from Bradypus tridactylus, Tamandua tetradactyla and Alouatta seniculus and two from Saguinus midas) were of a single zymodeme close to Trypanosoma rangeli reference stock RGB. This is the first confirmation of the presence of Tr. rangeli in French Guiana, and the first time that it has been identified, by iso-enzyme analysis, in the neotropical primates A. seniculus and S. midas. Two other stocks, isolated from Choloepus didactylus, were related to Endotrypanum schaudinni reference stock LEM 2790. Although the remaining stocks, one from C. didactylus and the other from A. seniculus, clustered together on UPGMA and in a Wagner tree, they did not appear to be related to any of the reference stocks included in the UPGMA dendrogram.

Electrophoretic patterns and immunoglobulin G levels in mouflon (Ovis orientalis musimon) and roe deer (Capreolus capreolus).

Serum protein and immunoglobulin G (IgG) levels were measured from 16 healthy mouflons (Ovis orientalis musimon) and from 28 roe deer (Capreolus capreolus) in French and Catalonian populations. Electrophoretic patterns were described for each species. The only significant gender-related difference was increased beta-globulins in French roe deer males. Significantly increased alpha1-globulin levels and decreased IgG levels occurred in the young Catalonian roe deer compared with adults from the same group. Values for total proteins, beta-globulins, gamma-globulin, and IgG were significantly higher in the French roe deer, while albumin and albumin/globulin ratio were higher in the Catalonian roe deer. Both populations had the same relative migration distances of the various protein fractions of the electrophoresis.

Genetic variation within the ticks Ixodes holocyclus and Ixodes cornuatus from south-eastern Australia.

Ticks from mainland Australia (Victoria, New South Wales and Queensland) and Tasmania, identified morphologically as either Ixodes holocyclus or Ixodes cornuatus, were compared genetically using 24 enzyme loci. The results showed that ticks from three localities in Victoria were genetically similar to I. cornuatus in Tasmania, but both groups had fixed genetic differences at >45% of loci compared with other ticks on the mainland. In addition, there were fixed genetic differences at 0-60% of loci among I. holocyclus from different localities on the mainland. Ixodes holocyclus samples could be divided into four distinct clusters (with fixed genetic differences >15%), three of which were represented by one or two specimens. Nonetheless, these electrophoretic data suggest that I. holocyclus represents a species complex. The results also showed that the morphological criteria used to identify specimens were not always accurate because several specimens had been mis-identified morphologically. Despite limitations with the morphological identification, this study has demonstrated that I. cornuatus can be distinguished from the I. holocyclus species complex using six enzyme loci, providing the foundation for a re-examination of morphological characteristics. The present study has shown that I. cornuatus and the I. holocyclus complexes have a greater distribution than previously reported, with both occurring in sympatry at Cape Patterson, on the southern coastline of Victoria.

Multiple myeloma in cats: variable presentation with different immunoglobulin isotypes in two cats.

Multiple myeloma was diagnosed in two cats with monoclonal hyperglobulinemia, proteinuria, and plasma cell proliferations in bone marrow. An immunoglobulin G-producing myeloma occurred in the vertebral bone marrow of one cat, and twice responded to surgical reduction followed by a combination of local irradiation and chemotherapy. The cat's survival time was approximately 2 years. The other myeloma in a cat that presented with hypercalcemia and renal insufficiency involved visceral organs and produced a biclonal peak due to immunoglobulin A dimer formation on serum electrophoresis. This cat's tumor did not respond to chemotherapy.

Use of multilocus enzyme electrophoresis to distinguish clinically important strains of Staphylococcus intermedius from the skin of dogs.

AIMS: To use multilocus enzyme electrophoresis to determine the genetic structure of Staphylococcus intermedius from normal skin of dogs and those isolated from a variety of disease conditions and to distinguish clinically important strains in dogs. METHODOLOGY: The diversity amongst 129 isolates of S intermedius from the skin and mucosa of 32 healthy dogs and 120 isolates from diseased sites in 120 individual dogs was examined using multilocus enzyme electrophoresis. Associations among ETs were examined to determine the diversity of isolates. RESULTS: Twenty two ETs were distinguished comprising 21 containing isolates from diseased sites and 11 containing isolates from normal dogs. The majority of isolates (171 of 249; 69% were located in two ETs (ET1 and ET 4), that were not distinguishable phenotypically. ET 1 contained 94 isolates (54 isolates from healthy dogs and 40 isolates from diseased sites) and ET 4 contained 77 isolates (46 from healthy dogs and 31 isolates from diseased sites). Further, 77.5% of isolates from healthy dogs were present in ET 1 and ET 4 and 59% of isolates from diseased dogs belonged to the same two ETs. There was only a small difference in genetic diversity among isolates taken from healthy dogs (11 ETs; H = 0.182) and those isolates taken from clinical specimens from diseased dogs (21 ETs; H = 0.218). Of the 21 ETs from diseased sites, ET 16 contained all six isolates from Staphylococcal Scalded Skin Syndrome in racing Greyhounds. CONCLUSIONS: The small difference in genetic diversity between isolates from the skin and mucosa of healthy dogs and isolates from various diseases, as well as the presence of the majority of isolates in two ETs, is consistent with the role of S intermedius as an opportunistic pathogen. The confinement of all Staphylococcal Scalded Skin Syndrome isolates within one ET is confirmation of this entity as a distinct disease of dogs.

Effects of sarcoptic mange on serum proteins and immunoglobulin G levels in chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica).

Three groups of chamois (Rupicapra pyrenaica) and three groups of Spanish ibex (Capra pyrenaica) were established to study the effects of sarcoptic mange on serum proteins and immunoglobulin G (IgG) levels. The first group of chamois consisted of 22 healthy Pyrenean chamois (R. pyrenaica pyrenaica) from a non-infested area, the second group consisted of 20 healthy Cantabrian chamois (R. p. parva) from an area where sarcoptic mange has been reported since 1994 and the third group consisted of 16 Cantabrian chamois from the same area but naturally infested by Sarcoptes scabiei. The first group of Spanish ibex was 39 healthy animals from a sarcoptic mange non-infested area, the second group was 23 healthy animals from a sarcoptic mange infested area and the third group consisted of 20 animals from the same area but naturally infested with the parasite. Blood samples were taken after killing the animals as part of hunting programmes. Values for total proteins, gamma-globulin and IgG were higher in infested and healthy chamois from the infested area compared to healthy chamois from the non-infested area, and IgG levels were higher in infested chamois compared to healthy-exposed chamois. Values for alpha2-globulin were higher in healthy Cantabrian chamois. In Spanish ibex, albumin, alpha2-globulin and IgG levels were lower in the healthy Spanish ibex from the non-infested area than in healthy animals from an infested area. The differences found in the chamois were indicative of the establishment of a humoral antibody response in the animals in contact with the disease. As the IgG levels were not significantly different between healthy and infested Spanish ibex from the same area, a different pattern of chronic infection with humoral response to the disease was suggested.