Renato Zanatta

In vitro evaluation of canine hemostasis following dilution with hydroxyethyl starch (130/0.4) via thromboelastometry.

OBJECTIVE To assess the effects of in vitro hemodilution of canine blood with a low molecular weight hy-droxyethyl starch (HES 130/0.4) by means of thromboelastometry (TEM). DESIGN In vitro experimental study. SETTING University Teaching Hospital. ANIMALS Ten healthy adult staff-owned dogs were sampled for the purposes of the study. Samples were also collected from 45 clinically normal dogs to establish thromboelastometric reference intervals. INTERVENTIONS For each dog whole blood was collected by jugular venipuncture and placed into tubes containing 3.8% trisodium citrate and stored at 37°C. Two standard dilutions (1:4 and 1:10) from canine blood specimens were prepared with HES 130/0.4 and saline 0.9%. MEASUREMENTS AND MAIN RESULTS The effects of dilution were studied by TEM. No statistically significant differences were observed between the TEM values of the control samples and the samples diluted with saline solution (ratio 1:10 and 1:4). In contrast, hypocoagulability was observed in the samples treated with HES 130/0.4, with more severe effects at 1:4 dilution than at 1:10 dilution. The 1:4 dilution with HES 130/0.4 produced marked alterations: CT (P = 0.035) and CFT (P = 0.0007) on the ex-TEM profile, CT (P = 0.0005) and ML (P = 0.0017) on the fib-TEM profile and CFT (P = 0.0043) on the in-TEM, were all significantly increased (P < 0.05), whereas MCF (P = 0.0042) and alpha angle (P = 0.002) on the in-TEM and MCF (P = 0.011) and alpha angle (P = 0.001) ex-TEM profiles and MCF (P = 0.0001) on the fib-TEM profile were significantly decreased (P < 0.05). CONCLUSIONS Dilution of canine blood with HES 130/0.4 results in a thromboelastometric pattern consistent with hypocoagulation and this effect appears to result from a dose-dependent alteration in fibrinogen concentration and inhibition of platelet function. As this is an in vitro study, further in vivo investigations are necessary to confirm the results.

Prognostic Indicators for Dogs with Dilated Cardiomyopathy

The purpose of this study was to investigate the prognostic value of various clinical, ECG, echocardiographic, and Doppler echocardiographic variables in dogs with dilated cardiomyopathy. The relationship to survival of 11 variables was evaluated in 63 dogs. Studied variables were age at time of diagnosis, class of heart failure (HF), dyspnea, ascites, atrial fibrillation (AF), ejection fraction (EF), E-point septal separation, end-diastolic volume index, end-systolic volume index (ESV-I), and restrictive or nonrestrictive transmitral flow (TMF) pattern. Median survival time was 671 days (lower 95% confidence limit, 350 days). Survival curves showed that severity of HF, ascites, ESV-I greater than 140 mL/m2, EF less than 25%, and restrictive TMF pattern had a significant negative relation to survival time. Thirty-nine dogs with both sinus rhythm and AF presented adequate TMF recordings; in these dogs, after stratification by TMF pattern, the restrictive TMF pattern was the most important negative prognostic indicator. We conclude that in dogs with dilated cardiomyopathy the restrictive TMF pattern appears to represent a useful prognostic indicator. Class of HF, ascites, ESV-I, and EF are also useful indexes if an adequate TMF pattern is not recorded.

Diagnostic and prognostic value of serum lactate dehydrogenase (LDH) and LDH isoenzymes in canine lymphoma

In humans, serum lactate dehydrogenase (LDH) is used as a prognostic indicator in non-Hodgkin’s lymphoma (NHL). In particular, elevated values of LDH (Schneider et al., 1980) and increases of the isoenzymes LDH2 and LDH3 (Dumontet et al., 1999) have been correlated with shortened survival times. In another study, the same isoenzymes were elevated in patients affected by latent NHL (Rotemberg et al., 1984). However, in veterinary medicine only a very limited amount of information is available regarding the pattern of LDH isoenzymes related to lymphoma. An increase in the value of serum LDH has been reported in dogs affected by leukaemia (Leifer and Matus, 1986) and in cats with large granular lymphoproliferative disorder (Buracco et al., 1992). In canine lymphomas, however, similar correlations between high concentrations of serum LDH and clinical staging or survival times have not been found (Greenlee et al., 1990). Bezzecchi and colleagues (1979) found that an increase in LDH3 was a possible indicator of latent disease. In an earlier study by our group, we showed that LDH2 and LDH3 were increased in dogs with lymphoma (Abate et al., 1997). The present report evaluates the clinical utility of monitoring changes in total and isoenzymatic patterns of LDH at diagnosis and during the course of therapy.

Membranoproliferative Glomerulonephritis Type III in a Simultaneous Infection of Leishmania Infantum and Dirofilaria Immitis in a Dog

In this report a 9-year-old female German Shepherd dog with a membranoproliferative glomerulonephritis (MPGN) type III associated with concomitant infection of Dirofilaria immitis and Leishmania infantum is presented. Light microscopic evaluation of kidney revealed a diffuse hypercellularity and thickening of glomerular basement membrane. Heavy and coarse granular complement C3 deposition and a weaker positive reaction to immunoglobulin G were present along peripheral glomerular basement membrane and in the mesangium in the immunofluorescent study. Transmission electron microscopy revealed deposits in the mesangium, subendothelium, and subepithelium. These lesions are compatible with membranoproliferative glomerulonephritis type III in humans.

Serum Values of Cardiac Troponin-T in Normal and Cardiomyopathic Dogs

Together with tropomyosin, the troponin complex participates in the contraction of skeletal and cardiac muscles. Troponin is composed of three subunits, namely troponin C (TnC), troponin I (TnI), and troponin T (TnT), which binds tropomyosin. There are three different isoforms of TnT and TnI, two of which are specific for skeletal muscle, while the third is specific for the myocardium (cTnT and cTnI). As the homology between troponins is about 95% among mammals, commercial diagnostic kits designed for use in humans also provide excellent results in other animals. (O’Brien et al., 1997). In human medicine, the measurement of cTnT or cTnI levels is now used together with measurement of the levels of creatine kinase (CK) and lactate dehydrogenase (LDH) and their isozymes for assessment of myocardial damage. For diagnostic purposes, one advantage is that these proteins are normally absent in blood in subjects without myocardial damage. Even minimal increases in the serum levels of these proteins are indicative of myocardial damage and, in the case of cTnT, it is detectable in serum from 2 h to 14 days following the onset of myocardial damage. Moreover, in contrast to the isozymes of CK and LDH, the results are not influenced by use of different clinical samples (hemolytic, lipemic, bile samples). The degree of increase in the level of cTnT is relatively proportional to the degree of myocardial damage, with prognostic implications as well. At the same time, the extreme sensitivity of this technique also allows for its use in other nonischemic pathologies including chronic cardiac insufficiency, pulmonary emboli, acute myocarditis, systemic hypertension, arrhythmias, and toxicity from doxorubicin (De Francesco, 2002). In veterinary medicine, O’Brien et al. (1997) have demonstrated the utility of measuring the levels of cTnT in the diagnosis of experimentally induced myocardial damage.

Corynebacterium urealyticum urinary tract infection in a cat with urethral obstruction

Corynebacterium urealyticum is an uncommon cause of urinary tract infections in cats. However, it is difficult to diagnose and if left untreated it may result in irreversible bladder lesions. C urealyticum is a multiantibiotic-resistant bacterium whose culture requires special care. Risk factors for the occurrence of this infection include urological procedures, foreign bodies, bladder mucosa abnormalities, immuno-suppressed states and antibiotic treatment. This report describes an unusual case of C urealyticum urinary infection in a young cat with pre-existing urethral obstruction. C urealyticum was isolated in pure cultures from two urine samples. Clinical and ultrasound features, results of the urinalysis and urine culture are described as well as therapeutic treatment and eventual favourable outcome to treatment with amoxycillin–clavulanic acid.

Severe renal failure in a dog resembling human focal segmental glomerulosclerosis.

A case of renal disease in a dog resembling human focal segmental glomerulosclerosis is presented. A kidney biopsy from this animal showed focal glomerular sclerosis, with variable distribution, affecting the perihilar and peripheral segments of the glomerular tuft. Non-sclerotic glomeruli were markedly enlarged. Interstitial fibrosis in association with tubular atrophy affected approximately 20% of the area of the biopsy. Immunofluorescence labelling showed immunoglobulin M deposits entrapped in segmental sclerotic areas and ultrastructural examination revealed segmental sclerosis and obliteration of capillaries, vacuolation of podocytes and diffuse effacement of foot processes. The dog was humanely destroyed 1 month later. At necropsy examination there was severe end-stage kidney disease with interstitial fibrosis involving more than 60% of the renal tissue. The clinical course and the microscopical, immunofluorescence and ultrastructural findings in this case have similarity to focal segmental glomerulosclerosis in man.

Genetic and phenotypic characterisation of Escherichia coli producing cefotaximase-type extended-spectrum β-lactamases: first evidence of the ST131 clone in cats with urinary infections in Italy

The incidence of cefotaximase (CTX-M)-type extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has increased dramatically in humans and animals since the middle of the last century. E coli that produce CTX-M β-lactamase represent a major cause of urinary tract infections, and pose a significant therapeutic challenge to both human and veterinary medicine. As data on uropathogenic CTX-M-producing strains in cats are limited, the aim of this study was to describe the genetic character and antibiotic resistance phenotypes of CTX-M-producing E coli isolated from cats with cystitis. Seven of 15 E coli bacteria isolated from 138 urine samples had the CTX-M gene and were therefore included in this study. These isolates were screened by polymerase chain reaction for the presence of 14 extra-intestinal virulence factors, class 1 and class 2 integrons, and to identify their phylogenetic groups. Multi-locus sequence typing (MLST) of the strains and susceptibility testing (disc diffusion method) were also performed. Virulence factor iutA was the most frequent determinant identified (86.7%), and the majority of CTX-M-producing strains (n = 5) carried class 1 integrons. MLST allowed us to discriminate four known sequence types (ST131, ST555, ST602, ST155) and three novel sequence types (ST3847, ST3848, ST4181). To the best of our knowledge, this is the first study to report uropathogenic CTX-M-producing E coli ST131 in cats in Italy. Accurate diagnostics and prudent use of antimicrobials are recommended to avoid the spread of multidrug-resistant pathogens in veterinary medicine and to prevent their transmission to humans.

Bilateral juvenile renal dysplasia in a Norwegian Forest Cat

Renal dysplasia is defined as a condition of disorganised development of renal parenchyma due to abnormal differentiation. The case of a 5-month-old intact male Norwegian Forest Cat with a history of polyuria and polydipsia is reported. Ultrasonographic examination showed a slight enlargement of kidneys. Biochemical parameters, haematological examinations and clinical signs were compatible with chronic renal failure (CRF). Histological examination was correlated with a primary tubular disorganisation and modification of glomerular compartment. The clinical history together with the histological lesions is consistent with bilateral juvenile renal dysplasia in this cat. To our knowledge, feline renal dysplasia has been reported in fetal infections with panleukopenia virus; no reports indicate the idiopathic origin in feline dysplastic lesions.

Validation of a Recombinant Based Antibody ELISA for Diagnosis of Human and Canine Leishmaniasis

Abstract In this study, a recombinant chimeric antigen (CA) ELISA was validated as a single test for both human and dog leishmaniasis. Serum panels included 327 human and 339 canine IFAT-positive and 1113 human and 1078 canine IFAT-negative samples. CA-ELISA was carried out using the same serum dilution, and labelled protein A as secondary reagent. Test performances were calculated using ROC analysis. For the human panel, the test showed diagnostic accuracy (DA) 0.974, specificity (Sp) 97.12%, sensitivity (Se) 91.44%, and concordance (K) 0.88. The dog panel showed DA 0.998, Sp 99.54%, Se 98.54%, and K 0.98. The proposed method is the best recombinant antigen-based ELISA, and can be used as IFAT substitute for mass screening.