Leo Brunnberg

Use of C‐reactive protein to predict outcome in dogs with systemic inflammatory response syndrome or sepsis

BACKGROUND There is a high mortality rate in patients with systemic inflammatory response syndrome (SIRS) or sepsis. Therefore, an early diagnosis and prognostic assessment is important for optimal therapeutic intervention. The objective of the study was to evaluate if baseline values and changes in serum C-reactive protein (CRP) might predict survival in dogs with SIRS and sepsis. DESIGN Prospective study; July 2004 to July 2005. SETTING Small Animal Clinic, Berlin, Clinic of Small Animal Medicine, Munich. ANIMALS Sixty-one dogs. MEASUREMENTS AND MAIN RESULTS For the CRP analysis blood was drawn on day 0, 1, and 2; CRP was measured using a commercial ELISA test kit. Thirteen dogs suffered from nonseptic SIRS and 48 dogs from sepsis. The 14-day survival rate was 61% (69% nonseptic SIRS, 58% sepsis). Serum CRP was higher in sick dogs compared with controls (P<0.001). Over the 3-day period surviving dogs (n=31) displayed a significantly greater decrease in CRP than nonsurvivors (n=10) (P=0.001). No correlation was found between the initial CRP concentrations and the survival rate. The changes in CRP corresponded to the survival rate (P=0.01). CONCLUSION There was no significant relationship between the survival rate in dogs with nonseptic SIRS or sepsis and the initial serum CRP concentrations. There was a correlation between decreasing CRP concentrations and recovery from disease. However, the changes in CRP concentrations over a 3-day period correctly predicted survival in 94% of dogs and death in 30% of the dogs (false positive rate 22%).

Genetic damage in operating room personnel exposed to isoflurane and nitrous oxide

OBJECTIVES: To evaluate genetic damage as the frequency of sister chromatid exchanges and micronuclei in lymphocytes of peripheral blood of operating room personnel exposed to waste anaesthetic gases. METHODS: Occupational exposure was measured with a direct reading instrument. Venous blood samples were drawn from 10 non-smokers working in the operating room and 10 non-smoking controls (matched by age, sex, and smoking habits). Lymphocytes were cultured separately over 72 hours for each assay with standard protocols. At the end of the culture time, the cells were harvested, stained, and coded for blind scoring. The exchanges of DNA material were evaluated by counting the number of sister chromatid exchanges in 30 metaphases per probe or by counting the frequency of micronuclei in 2000 binucleated cells. Also, the mitotic and proliferative indices were measured. RESULTS: The operating room personnel at the hospital were exposed to an 8 hour time weighted average of 12.8 ppm nitrous oxide and 5.3 ppm isoflurane. The mean (SD) frequency of sister chromatid exchanges was significantly higher (10.2 (1.9) v 7.4 (2.4)) in exposed workers than controls (p = 0.036) the proportion of micronuclei (micronuclei/500 binucleated cells) was also higher (8.7 (2.9) v 6.8 (2.5)), but was not significant (p = 0.10). CONCLUSION: Exposure even to trace concentrations of waste anaesthetic gases may cause dose-dependent genetic damage. Concerning the micronuclei test, no clastogenic potential could be detected after average chronic exposure to waste anaesthetic gas. However, an increased frequency of sister chromatid exchanges in human lymphocytes could be detected. Although the measured differences were low, they were comparable with smoking 11-20 cigarettes a day. Due to these findings, the increased proportion of micronuclei and rates of sister chromatid exchanges may be relevant long term and need further investigation.

Quantification of cytokines and inflammatory mediators in a three-dimensional model of inflammatory arthritis.

Human recombinant IL-1beta and TNFalpha have been previously used to induce a cytokine response in canine chondrocytes. In order to establish this functional relation in a homologous system in vitro, we have developed both 2D and 3D models of inflammatory arthritis using canine recombinant cytokines in canine articular chondrocytes. IL-1beta and TNFalpha were cloned and subsequently expressed in Escherichia coli. The purified recombinant canine cytokines were used to simulate inflammation in vitro and the expression of typical inflammation markers such as proinflammatory cytokines (IL-1beta, IL-6, IL-8, GM-CSF and TNFalpha), enzyme mediators (MMP-3 MMP-13, iNOS, COX-2) and their catabolites (NO, PGE(2)) was measured. High expression of proinflammatory cytokines, enzyme mediators and their catabolites was only observed in IL-1beta/TNFalpha stimulated cells. We conclude that the canine IL-1beta and TNFalpha generated in this study are biologically active and equally effective in the canine cell culture systems. Inducing an inflammatory pathway by canine exogenous cytokines in canine chondrocytes provides a useful tool for the study of canine inflammatory arthritis.

Determination of plasma albumin concentration in healthy and diseased turtles: a comparison of protein electrophoresis and the bromcresol green dye‐binding method

BACKGROUND In reptile medicine, plasma chemistry analysis is widely used for the evaluation of an individuals health status. The standard method for the determination of plasma albumin concentration is protein electrophoresis combined with the determination of total protein concentration, but the bromcresol green (BCG) dye-binding method is also used. The reliability of the BCG method for the measurement of albumin concentration in reptiles is unknown. OBJECTIVE The aim of this study was to compare the plasma albumin values of turtles obtained by protein electrophoresis and the BCG method. METHODS Between March 2008 and September 2008, heparinized plasma samples from 16 clinically healthy and 10 diseased turtles of different species were collected. Plasma albumin concentrations were measured by protein electrophoresis and by the BCG method. The results of the 2 methods were compared using Passing-Bablok regression and Bland-Altman plots. RESULTS Albumin concentration measured by BCG was weakly correlated with the corresponding protein electrophoretic values in all turtles (r(s)=.610, P<.001) and in healthy turtles evaluated separately (r(s)=.700, P=.003), whereas in diseased turtles no such correlation was found (r(s)=.374, P=.287). The albumin concentration measured with the 2 different methods differed significantly in all turtles (P=.009; Wilcoxons test) and in healthy turtles (P=.005) but not in diseased animals (P=.241). In the Bland-Altman plot a systematic error was found between the 2 methods in diseased turtles. CONCLUSION Measurement of albumin by the BCG dye-binding method may lead to inaccurate results for plasma albumin concentration, especially in ill turtles. Therefore, for health assessment in turtles, albumin should be measured by protein electrophoresis.

Staphylococcus aureus and MRSA colonization rates among personnel and dogs in a small animal hospital: association with nosocomial infections

The genetic relationship of thirty Methicillin-resistant S. aureus (MRSA) isolates derived from the nasal cavities of canine patients hospitalized (n = 7), veterinary personnel (n = 20), and environmental sources (n = 3) sampled during a 20-month investigation period, were analyzed in this study. Genetic relatedness of all MRSA isolates was investigated involving commonly used typing techniques: Pulsed-field gel electrophoresis (PFGE), Multilocus sequence typing (MLST), spa typing, PCR for detection of Panton-Valentine leukocidine (PVL) genes and staphylococcal cassette chromosome mec-typing (SCCmec). Analysis of typing results revealed a certain predominant (72%) genotype: PFGE type IMT-A, ST22, spa type t032, SCCmecIV. This genotype has been reported previously (Walter et al., 2008c) being the predominant PFGE type associated with MRSA-positive clinical specimens, mostly from wound infections, derived from small and exotic animals of that facility. Furthermore, occasionally high rates in nasal colonization of veterinary personnel (18 of 88: 20%) in one of three personal screening periods were accompanied by a sudden rise of MRSA infections in small animals. Our data strongly indicate that high rates of colonized veterinary staff lead to an increase of nosocomial infections in small animal hospitals. We therefore recommend the introduction of surveillance of nosocomial infections especially in surgical veterinary hospitals.

Influence of kidney function on urinary excretion of albumin and retinol-binding protein in dogs with naturally occurring renal disease

OBJECTIVE To evaluate excretion of urinary albumin (UAlb) and urinary retinol-binding protein (URBP) in dogs with naturally occurring renal disease. ANIMALS 64 client-owned dogs. PROCEDURES Dogs were assigned to groups according to plasma creatinine concentration, urinary protein-to-urinary creatinine ratio (UP:UC), and exogenous plasma creatinine clearance (P-Cl(Cr)) rates: group A (n = 8), nonazotemic (plasma creatinine < 125 μmol/L) and nonproteinuric (UP:UC < 0.2) with P-Cl(Cr) rate > 90 mL/min/m²; group B (26), nonazotemic and nonproteinuric with P-Cl(Cr) rate 50 to 89 mL/min/m²; group C (7), nonazotemic but proteinuric with P-Cl(Cr) rate 53 to 98 mL/min/m²; group D (8), azotemic and borderline proteinuric with P-Cl(Cr) rate 22 to 45 mL/min/m²); and group E (15), azotemic and proteinuric (P-Cl(Cr) not evaluated). The UAlb and URBP concentrations were measured via ELISA; UAlb-to-urinary creatinine (UAlb:UC) and URBP-to-urinary creatinine (URBP:UC) ratios were determined. RESULTS UAlb:UC and URBP:UC did not differ between groups A and B. Increased UAlb: UCs and URBP:UCs were paralleled by increased UP:UCs in groups C, D, and E relative to values from groups A and B, independent of azotemia. There were significant positive correlations of UP:UC with UAlb:UC and of UAlb:UC with URBP:UC (r = 0.82 and 0.46, respectively). However, UP:UC, UAlb:UC, and URBP:UC were not significantly correlated with P-ClCr rate. CONCLUSIONS AND CLINICAL RELEVANCE UAlb and URBP concentrations were paralleled by urinary protein concentrations and may be useful in assessing renal management of plasma proteins. Determination of urinary protein, UAlb, or URBP concentration was not sufficiently sensitive to detect reduced P-Cl(Cr) in nonazotemic dogs.

Serum thyroxine concentrations in clinically healthy pet guinea pigs (Cavia porcellus).

BACKGROUND Guinea pigs (Cavia porcellus) are often presented as patients in veterinary practice. Nevertheless, only limited information is available about endocrine diseases or thyroxine reference values for the species. OBJECTIVE The aim of this study was to determine serum thyroxine concentrations in a well-defined population of clinically healthy pet guinea pigs. METHODS Between October 2007 and July 2008, serum samples were collected from 40 clinically healthy guinea pigs of different sexes, ages, and breeds that were presented to our clinic for a general health check or for castration. Pregnant females were excluded from the study. Thyroxine concentration was measured using a chemiluminescence test (Immulite 2000 Canine Total T4). RESULTS Thyroxine concentrations ranged from 14.2 to 66.9 nmol/L (1.1-5.2 microg/dL) with a median value of 27.0 nmol/L (2.1 microg/dL). Females (n=16) had significantly (P=.039; Mann-Whitney U-test) lower thyroxine values than castrated males (n=8), whereas no differences were found between females and intact males (n=16) or between intact and castrated males. No significant correlation was found between thyroxine concentration and age. CONCLUSION This is the first report of serum thyroxine reference values for a well-defined population of healthy pet guinea pigs as measured by a chemiluminescence assay. The results were higher than those previously reported for this species and emphasize the importance of using appropriate reference intervals for the diagnosis of hyperthyroidism.

Surgical approach for tentorial meningiomas in cats: a review of six cases.

The surgical technique for removal of tentorial meningiomas is described on six cats using a unilateral temporal supracerebellar transtentorial approach. Complete gross tumour resection was achieved in four of six cats. In one cat, only subtotal resection was achieved. One cat died shortly after surgery because of extensive cerebral haemorrhage. The surgical approach, combined with cisternal or ventricular cerebrospinal fluid puncture and an open-window technique (tumour fenestration and enucleation) provided sufficient visibility and tumour accessibility without excessive manipulation of the brain parenchyma. In all patients, a postoperative transient worsening of the clinical signs was observed. The neurological signs resolved with time with the exception of blindness in two cats. All five surviving cats were monitored for a mean follow-up time of 19 months (median 20 months; range 6–30 months). All patients died or were euthanased because of tumour regrowth within the follow-up period. Although challenging, surgical treatment is a useful therapeutic measure in the treatment of cats presenting with tentorial meningiomas.

Encephalitis in a rabbit caused by human herpesvirus-1.

CASE DESCRIPTION An 8-month-old sexually intact male rabbit was examined because of a 2-day history of anorexia, epiphora of the left eye, bruxism, hypersalivation, and ataxia. CLINICAL FINDINGS Physical examination of the rabbit revealed bilateral conjunctivitis, hypersalivation, and severe signs of CNS dysfunction such as incoordination, intermittent myoclonic seizures, and opisthotonus. Results of hematologic and serum biochemical analyses revealed only lymphopenia, a relative monocytosis, and an increase in serum activity of creatine phosphokinase and serum concentration of total protein. Serum antibodies against Encephalitozoon cuniculi and Toxoplasma gondii were not detected. TREATMENT AND OUTCOME Despite IV administration of crystalloid fluids and treatment with antimicrobials, vitamin B complex, nutritional support, and prednisolone, the condition of the rabbit deteriorated; it was euthanized 7 days after admission. Histologic evaluation of brain tissue revealed lesions characteristic of severe, diffuse, nonsuppurative meningoencephalitis and a few large, eosinophilic, intranuclear inclusion bodies in neurons and glial cells. The DNA of human herpesvirus-1 was detected in the nuclei of glial cells, lymphocytes, and neurons by means of in situ hybridization. The rabbits owner, who reported having had a severe labial and facial herpesvirus infection 5 days before the onset of clinical signs in the rabbit, was suspected to be the origin of infection for the rabbit. CLINICAL RELEVANCE Human herpesvirus-1 may be transmissible from humans to rabbits, and infection with this virus should be considered as a differential diagnosis in rabbits with CNS signs of disease.

Evaluation of a point-of-care test for canine C-reactive protein.

BACKGROUND In veterinary medicine, there is increasing interest in measuring C-reactive protein (CRP) as a tool for diagnosis and monitoring of inflammatory diseases. Reported CRP concentrations for healthy dogs have ranged from 0 to 8.9 mg/L. OBJECTIVES The aims of this study were to evaluate a canine-specific point-of-care (POC) lateral flow immunoassay for qualitative CRP measurement in healthy and diseased dogs and to compare results with those obtained by a quantitative ELISA. METHODS Blood samples from 73 client-owned dogs were available for testing: 16 healthy dogs and 57 dogs with a variety of infectious, inflammatory, or neoplastic diseases. CRP was measured in heparinized whole blood samples and serum with the TECO medical Dog CRP-visual POC test. A red line develops in the POC device if CRP is ≥ 5 mg/L, and results are scored as negative or positive. An ELISA validated previously for canine serum was used as the reference method. RESULTS For all dogs, serum CRP concentrations measured by the ELISA ranged from 0.1 to ≥ 350 mg/L (median = 38 mg/L). Percentages of the CRP POC test results that agreed with the ELISA results were 98.6% for whole blood and 97.3% for serum samples. For serum samples, sensitivity of the POC test was 96.4% and specificity was 81.3%. For whole blood, sensitivity was 94.7% and specificity was 93.8%. CONCLUSIONS The POC test had very good agreement with the ELISA test and had high sensitivity and specificity; therefore, it can be used as a qualitative test to screen for increases in CRP concentrations.