Age-related alterations to immune parameters in Labrador retriever dogs

In order to assess age-related changes in the immune status of Labrador retriever dogs, leukocyte phenotypes, lymphocyte proliferative capacity, and serum antibody levels were measured in four cohorts of dogs, ranging from 2 to 10 years of age. Absolute numbers of white blood cells, lymphocytes, monocytes, granulocytes, and CD3+, CD4+, CD8+ and CD21+ lymphocytes significantly decreased with increasing age. Relative percentages of lymphocytes and CD4 cells were significantly decreased, and relative percentages of granulocytes and CD8 cells significantly increased, with age. The CD4:CD8 ratio showed a significant age-related decrease. Proliferative responses of T-cells to mitogens in whole-blood cultures either increased (Concanavalin A) or remained the same (phytohemagglutinin) with age when data was normalised to allow for differences in responding cell number. Similarly, normalised data of proliferative response to anti-CD3 stimulation together with phorbol 12-myristate 13-acetate showed an age-related increase. Serum levels of total IgA significantly increased with age whereas total IgG levels remained unchanged. These observations illustrate a significant change to a number of immune parameters with age. However, further work is required to determine whether the differences reported here are sufficient to cause overt or functional immune senescence in Labrador retriever dogs.     

Identification of putative adhesin genes in shigatoxigenic Escherichia coli isolated from different sources

Shiga toxin-producing Escherichia coli (STEC) is an important pathogen responsible for severe human intestinal and systemic infections. The bacterial factors required for colonization of the hosts are still not well defined. In this study, the prevalence of seven putative adhesive genes that are not encoded in the locus of enterocyte effacement (LEE) in 74 STEC strains isolated from humans (n=39), food (n=6), cattle (n=11), and pigs (n=18) was investigated by PCR. In addition, Shiga toxin (stx) and intimin (eaeA including alpha, beta, gamma, delta, epsilon, zeta variants) genes were tested. The most prevalent adhesin was that encoded by toxB gene (52 of 74 isolates; 70.3%). This marker was found in all 12 strains of O157:H7 serotype and in 23 of 32 (71.9%) isolates of the O157:NM serogroup. Moreover, this gene was also present in other 17 STEC of the non-O157 serogroup. The second most prevalent adhesin was that encoded by the lpfAO157/OI-154 gene (43 isolates; 58.1%). This marker was detected in LEE-positive strains of the O157 serogroup but also in 9 LEE-negative isolates of porcine origin. Several STEC isolates tested (42 strains; 56.7%) had the efa1 gene of the Efa1 putative adhesive marker. This adhesin was almost exclusively found among eaeA-positive strains recovered from humans, food and cattle. On the other hand, iha marker was detected either in LEE-positive (29 isolates) or LEE-negative (12 strains) STEC. Only two eaeA-negative strains had the saa putative adhesive gene. These results show that STEC strains may be able to express several putative adhesins. However, further studies are needed to evaluate the role of the genes identified in the present study in the pathogenesis of human infections.     

Idiopathic thrombocytopenia in Cavalier King Charles Spaniels

OBJECTIVE: To determine the prevalence of asymptomatic idiopathic macrothrombocytopenia in the population of Cavalier King Charles Spaniels (CKCS) in New South Wales (NSW) and to determine if it exhibits an autosomal recessive inheritance pattern. We also aimed to determine if significant differences existed when counting platelets manually, by auto analyser or by blood smear estimation in CKCS and mixed breed dogs. METHODS: Blood was collected from 172 dogs (152 CKCS and 20 mixed breed) and placed into sodium-citrate anticoagulant. Platelet counts were performed manually, by auto analyser and by blood smear estimates in CKCS and mixed breed dogs. Blood smears were also examined for platelet clumping and erythrocyte, leukocyte and platelet morphology. Pedigree analysis was performed to determine if an autosomal recessive inheritance pattern was supported. RESULTS: A statistically significant difference was found in platelet counts between CKCS and mixed breed dogs (P < 0.0001). CKCS had a platelet count that was 32% that of the controls (95% confidence interval, 28 to 37%). There was no significant difference between methods used to count platelets. Thirty percent of CKCS had macrothrombocytes. Pedigree analysis and examination of obtained and expected segregation ratios from 17 CKCS families supported an autosomal recessive pattern of Mendelian inheritance. CONCLUSIONS: A high prevalence of idiopathic macrothrombocytopenia exists in CKCS in NSW and automated or blood smear estimates are sufficient to count platelet numbers. Data supports an autosomal recessive inheritance pattern.     

Assessment of electrocardiographic parameters in healthy dogs undergoing dobutamine stress testing

The electrocardiographic effects of dobutamine stress testing (10 to 40 microg/kg/minute) were investigated in five conscious healthy dogs. We studied the changes in the duration and amplitude of P wave, PR interval, duration of QRS complex, R wave amplitude, QT interval, and heart rate. Development of arrhythmias and ST segment abnormalities were also recorded. It was observed that dobutamine significantly affects atrioventricular-nodal conduction and total electrical systole time at higher infusion rates. Only a single episode of sustained ventricular tachycardia was observed, which was promptly restored to sinus rhythm shortly after dobutamine infusion was discontinued. No ST segment abnormalities were detected. Dobutamine stress testing was concluded to play a role in some ECG parameters at higher infusion rates.     

Sarcomatoid renal cell carcinoma in a binturong (Arctictis binturong).

An adult, female binturong (Arctictis binturong) was examined due to lethargy, inappetence, and an abdominal mass. Diagnostic investigations, including radiographs, abdominal ultrasound, clinical laboratory findings, and a fine-needle aspirate of the mass, were suggestive of a sarcoma with metastasis. Necropsy and histopathologic findings confirmed a widely disseminated sarcomatoid variant of a renal cell carcinoma, likely originating in the left kidney, with metastasis to the right kidney, spleen, pancreas, liver, mesenteric lymph nodes, and lungs. This is the first report of this neoplasm in a binturong and only the second report in the veterinary literature. Sarcomatoid renal cell carcinoma is a rare histologic variant of renal cell carcinoma that is aggressive, commonly metastatic, and associated with a very poor prognosis in humans. Accurate antemortem diagnosis of this tumor may be complicated by its biphasic morphology, which may resemble carcinoma or sarcoma (or both), often necessitating the use of immunohistochemical techniques.     

Determination of biogenic amines in aerobically stored beef using high-performance thin-layer chromatography densitometry

The aim of this study was to develop a high-performance thin-layer chromatography densitometry method to simultaneously evaluate levels of different biogenic amines (BAs) in beef stored aerobically at 4°C. Steaks from M. longissimus thoracis et lumborum muscles were overwrapped with polyvinyl chloride film and stored for 0, 4, 7, and 10 days. For this purpose, the solvent system and pH for the derivatization of BAs with dansyl chloride were optimized. Moreover, this method was also validated for linearity, limits of detection and quantification, precision, and recovery. As found using this method in beef samples, spermine content decreased significantly after 7 days of storage, and tyramine and cadaverine were not detected until day 10.     

Paratuberculosis in cattle: a comparison of three serologic tests with results of fecal culture

Feces and blood were collected from cattle in 13 herds known to be infected with Mycobacterium paratuberculosis to evaluate a complement-fixation (CF) test, an agar gel immunodiffusion (AGID) test and an enzyme-linked immunosorbent assay (ELISA) for the serologic diagnosis of paratuberculosis. M. paratuberculosis was isolated from the feces of 36 of 192 cattle examined. Twenty-three culture-positive animals had CF test titers regarded as suspect or positive, 10 were positive by the AGID test and 34 were suspect or positive by the ELISA. Of the 156 culture-negative animals, the CF test agreed on 136, the ELISA on 129 and the AGID on 151.     

Effect of calcium-channel blockers and salbutamol on the isolated mare uterus — interaction with the calcium agonist Bay K 8644

The effects of nifedipine, verapamil and diltiazem were investigated in the isolated mare uterus in comparison with salbutamol. All the calcium-channel blockers and salbutamol inhibited the spontaneous, KC1- and electrically induced contractions; nifedipine and salbutamol were the most potent compounds. The calcium agonist Bay K 8644 (10(-8)-10(-6) mol/l) competitively antagonized the inhibitory effect of nifedipine (pA2 value = 8.54 +/- 0.06), whereas it was only slightly or totally ineffective against verapamil, diltiazem and salbutamol. These results indicate that calcium-channel blockers are potent inhibitors of mare uterine motility in vitro and emphasize the importance of Ca2+-related mechanisms in the control of uterine smooth-muscle contractility. Moreover, the validity of Bay K 8644 as a tool to distinguish classes of calcium-channel antagonists is confirmed.