International renal interest society best practice consensus guidelines for intermittent hemodialysis in dogs and cats.

Intermittent hemodialysis (IHD) is an advanced adjunctive standard of care for severe acute kidney injury (AKI) and other indications. Most animals with AKI are managed medically, however, when the disease is severe, medical management may not control the consequences of the disease, and animals with a potential for renal recovery may die from the consequences of uremia before recovery has occurred. Extracorporeal therapies aid the management of AKI by expanding the window of opportunity for recovery of sufficient kidney function to become dialysis independent. Intermittent hemodialysis (IHD) was introduced into veterinary medicine over 50 years ago, however, updated guidelines for the delivery of IHD have not been published for several decades. To that end, the International Renal Interest Society (IRIS) constituted a Working Group to establish best practice guidelines for the safe and effective delivery of IHD to animals with indications for dialytic intervention. The IRIS Working Group generated 60 consensus statements and supporting rational for a spectrum of prescription and management categories required for delivery of IHD on designated intermittent dialysis platforms (i.e., AKI, chronic hemodialysis and intoxications). A formal consensus method was used to validate the recommendations by a blinded jury of 12 veterinarians considered experts in extracorporeal therapies and actively performing IHD. Each vote provided a level of agreement for each recommendation proposed by the Working Group. To achieve a consensus, a minimum of 75% of the voting participants had to "strongly agree" or "agree" with the recommendation.

International Renal Interest Society best practice consensus guidelines for the diagnosis and management of acute kidney injury in cats and dogs.

Acute kidney injury (AKI) is defined as an injury to the renal parenchyma, with or without a decrease in kidney function, as reflected by accumulation of uremic toxins or altered urine production (i.e., increased or decreased). AKI might result from any of several factors, including ischemia, inflammation, nephrotoxins, and infectious diseases. AKI can be community- or hospital-acquired. The latter was not previously considered a common cause for AKI in animals; however, recent evidence suggests that the prevalence of hospital-acquired AKI is increasing in veterinary medicine. This is likely due to a combination of increased recognition and awareness of AKI, as well as increased treatment intensity (e.g., ventilation and prolonged hospitalization) in some veterinary patients and increased management of geriatric veterinary patients with multiple comorbidities. Advancements in the management of AKI, including the increased availability of renal replacement therapies, have been made; however, the overall mortality of animals with AKI remains high. Despite the high prevalence of AKI and the high mortality rate, the body of evidence regarding the diagnosis and the management of AKI in veterinary medicine is very limited. Consequently, the International Renal Interest Society (IRIS) constructed a working group to provide guidelines for animals with AKI. Recommendations are based on the available literature and the clinical experience of the members of the working group and reflect consensus of opinion. Fifty statements were generated and were voted on in all aspects of AKI and explanatory text can be found either before or after each statement.

Hyperlipasemia in dogs with acute kidney injury treated with and without hemodialysis.

BACKGROUND: Hyperlipasemia has been reported in dogs with acute kidney injury (AKI) but associations with AKI severity, hemodialysis (HD) treatment, and outcome have not been extensively evaluated. OBJECTIVES: Investigate the prevalence and clinical relevance of hyperlipasemia in dogs with AKI, treated with and without HD. ANIMALS: Client-owned dogs (n = 125) with AKI. METHODS: Retrospective data extraction from medical records, including signalment, cause of AKI, duration of hospitalization, survival, plasma creatinine concentration, and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methyresorufin) ester (DGGR) lipase activity at admission and throughout hospitalization. RESULTS: A DGGR-lipase activity >3× the upper reference limit (URL) was found in 28.8% and 55.4% of dogs at admission and during hospitalization, respectively, but only 8.8% and 14.9% of dogs, respectively, were diagnosed with acute pancreatitis. Hyperlipasemia >10 × URL was observed in 32.7% of dogs during hospitalization. The DGGR-lipase activity was higher in dogs with International Renal Interest Society (IRIS) Grades 4-5 than Grades 1-3, but correlation between DGGR-lipase activity and creatinine concentration was poor (rs = .22; 95% confidence intervals [CI], 0.04-0.38). Treatment with HD was not associated with DGGR-lipase activity independent of IRIS grade. Survival to discharge and 30 days after admission was 65.6% and 59.6%, respectively. High IRIS grades (P = .03) and high DGGR-lipase activity at admission (P = .02) and during hospitalization (P = .003) were associated with nonsurvival. CONCLUSIONS AND CLINICAL IMPORTANCE: Hyperlipasemia is frequent and often marked in dogs with AKI despite only a minority being diagnosed with pancreatitis. Hyperlipasemia is associated with AKI severity but not independently with HD treatment. High IRIS grade and hyperlipasemia were associated with nonsurvival.

The circulating renin-angiotensin-aldosterone system is down-regulated in dogs with glomerular diseases compared to other chronic kidney diseases with low-grade proteinuria.

Glomerular diseases (GD) lead to a variety of disorders of the vascular and the total body water volumes. Various pathomechanisms, including vascular underfill and overfill, have been suggested to explain these disturbances. Accordingly, the circulating renin-angiotensin-aldosterone system (cRAAS) is expected to be activated as either a cause or a result of these fluid disorders. The aim of this study was to characterize the activity of the cRAAS in dogs with GD and to evaluate its relationship with the vascular volume status. In a prospective study, we evaluated the plasma renin activity and the serum aldosterone concentration in 15 dogs with GD. Their fluid volume status was estimated with clinical variables reflecting volemia and hydration, echocardiographic volume assessment, N-terminal pro B-type natriuretic peptide, blood urea nitrogen:creatinine ratio, and the urinary fractional excretion of sodium. Ten dogs with chronic kidney disease (CKD) with matching degree of azotemia were recruited as controls. The activity of the cRAAS was low in 10 dogs, normal in 3 dogs, high in 1 dog and equivocal (high renin-low aldosterone) in 1 dog with GD. These dogs had a lower cRAAS activity than dogs with CKD (p = 0.01). The clinical evaluation showed 8 hypovolemic and 7 non-hypovolemic dogs; 3 dehydrated, 9 euhydrated and 3 overhydrated dogs. The cRAAS activity was not different between hypovolemic and non-hypovolemic dogs. The down-regulated cRAAS without obvious association with the clinical volume status of these dogs with GD, suggests different mechanisms of fluid volume dysregulation in dogs with GD than previously assumed. This finding however should be confirmed in a focused larger scale study, as it may influence the use of cRAAS blockers as part of the standard therapy of GD in dogs.

Diagnostic value of reticulocyte indices for the assessment of the iron status of cats with chronic kidney disease.

BACKGROUND: Reticulocyte indices have been suggested as alternatives to transferrin saturation (TSAT) for iron status assessment in humans and dogs but they have not been evaluated thoroughly in cats. OBJECTIVES: To assess the value of the reticulocyte indices for the diagnosis of iron deficiency in cats with chronic kidney disease (CKD) and chronic hematuria associated with subcutaneous ureteral bypasses (SUBs). ANIMALS: Sixty-four cats: 16 healthy, 14 CKD without SUB, and 34 CKD with SUB. METHODS: Prospective observational cross-sectional study of cats presented for routine nephrology visits. Primary outcomes included assessment of the diagnostic values of erythrocyte indices (mean corpuscular volume, hemoglobin, and hemoglobin concentration: MCV, MCH, and MCHC) and reticulocyte indices (mean corpuscular volume, MCVr; corpuscular hemoglobin, CHr), using TSAT as reference. RESULTS: Iron deficiency was diagnosed in 9/64 cats (14%). A receiver-operating characteristic curve analysis yielded a moderate discriminatory value for CHr in this diagnosis: area under the curve [AUC] = .75 (95% confidence interval, 0.48-0.89); P = .006; sensitivity 67%, specificity 82% for a cutoff of 15.9 pg. This compared favorably to MCVr (AUC = .63; P = .29), MCV (AUC = .58; P = .45), MCH (AUC = .64; P = .19), and MCHC (AUC = .7; P = .03). CONCLUSION AND CLINICAL IMPORTANCE: CHr added moderate value to the diagnosis of iron deficiency in cats with CKD.

Association of markers of endothelial activation and dysfunction with occurrence and outcome of pulmonary hemorrhage in dogs with leptospirosis.

BACKGROUND: Endothelial dysfunction might contribute to the development of leptospiral pulmonary hemorrhage syndrome (LPHS). HYPOTHESIS: Serum concentrations of markers of endothelial activation and dysfunction are higher in dogs with leptospirosis and correlate with the occurrence of LPHS and a higher case fatality rate. ANIMALS: Clinically healthy dogs (n = 31; 10/31 dogs confirmed healthy based on no detected abnormalities on blood work), dogs with leptospirosis with LPHS (n = 17) and without LPHS (n = 15), dogs with acute kidney injury not due to leptospirosis (AKI-nL, n = 34). METHODS: Observational study. Serum concentrations of soluble intercellular adhesion molecule 1 (sICAM-1), vascular endothelial growth factor (VEGF), and angiopoietin-2 (Ang-2) at admission were compared between groups. Correlations with outcome and the accuracy to predict LPHS were examined. RESULTS: Soluble intercellular adhesion molecule (sICAM-1), VEGF, and Ang-2 concentrations were higher in dogs with AKI-nL (sICAM-1 34.7 ng/mL, interquartile range [IQR] = 24.4-75.5; VEGF 43.1 pg/mL, IQR = 12.3-79.2; Ang-2 8.5 ng/mL, IQR = 6.2-12.3), leptospirosis without LPHS (sICAM-1 45.1 ng/mL, IQR = 30.6-59.0; VEGF 32.4 pg/mL, IQR = 12.5-62.6; Ang-2 9.6 ng/mL, IQR = 6.9-19.3), and LPHS (sICAM-1 69.7 ng/mL, IQR = 42.1-89.1; VEGF 51.8 pg/mL, IQR = 26.3-96.7; Ang-2 8.0 ng/mL, IQR = 5.6-12.2) compared to controls (P < .001). In dogs with leptospirosis, VEGF and sICAM-1 were higher in nonsurvivors (sICAM-1 89.4 ng/mL, IQR = 76.5-101.0; VEGF 117.0 pg/mL, IQR = 90.3-232.4) than survivors (P = .004) and sICAM-1 predicted the development of LPHS. CONCLUSIONS: Soluble intercellular adhesion molecule 1, VEGF, and Ang-2 do not discriminate leptospirosis from AKI-nL. In dogs with leptospirosis, sICAM-1 and VEGF predict outcome and sICAM-1 might identify dogs at risk for LPHS.

Evaluation of membrane-based therapeutic plasma exchange as adjunctive treatment for immune-mediated hematologic disorders in dogs.

BACKGROUND: Therapeutic plasma exchange (TPE) has been used increasingly to treat immunological diseases in dogs, although data concerning its efficacy are lacking. HYPOTHESIS/OBJECTIVES: To describe the clinic and the outcome of dogs with immune-mediated hematological disorders (IMHD) treated with membrane filtration TPE. ANIMALS: One hundred forty-six dogs diagnosed with IMHD, including 17 dogs treated with and 129 control dogs treated without TPE. METHODS: Retrospective study of cases selected with a search of all medical records for dogs diagnosed with IMHD (2010-2019). Primary outcomes included the assessment of safety and efficacy of adjunctive TPE compared to medical treatment alone. RESULTS: The TPE group included 7 dogs with immune-mediated hemolytic anemia (IMHA), 5 dogs with immune-mediated thrombocytopenia (IMT), and 5 dogs with combined IMHA-IMT; the control group included 63 dogs with IMHA, 47 dogs with IMT, and 19 dogs with IMHA-IMT. Dogs treated with TPE were more often refractory to initial immunosuppression (71%) than controls (43%; P = .04). Complications were observed in 15/43 TPE treatments (35%). The response rate of dogs treated with TPE was 83% in IMHA (controls, 65%; P = .5) and 80% in IMT (controls, 70%; P = .71). Overall, 12/17 dogs (71%) treated with TPE reached complete remission, 2/17 (12%) partial remission, and 3/17 (18%) died or were euthanized. Eighty-two percent of the dogs treated with TPE survived to discharge (controls, 69%; P = .4). CONCLUSIONS AND CLINICAL IMPORTANCE: Despite a bias toward dogs refractory to initial immunosuppression, dogs treated with adjunctive TPE had a similar outcome as dogs treated medically.

Evaluation of changes in the epidemiology of leptospirosis in dogs after introduction of a quadrivalent antileptospiral vaccine in a highly endemic area.

BACKGROUND: Since 2003, a marked increase in leptospirosis serogroup Australis has been observed in dogs in Switzerland. In 2013, a new quadrivalent antileptospiral vaccine (L4) was introduced, adding serogroups Australis and Grippotyphosa to Canicola and Icterohaemorrhagiae of the previous bivalent vaccines (L2). OBJECTIVE: To examine whether introduction of L4 was associated with decreased incidence of leptospirosis and decreased odds for dogs with acute kidney injury (AKI) to be diagnosed with leptospirosis. ANIMALS: Four hundred and sixty-nine dogs with AKI presented to a referral hospital, including 269 dogs with leptospirosis and 200 controls with other causes. METHODS: Descriptive section: disease incidence was evaluated for 3 consecutive periods: before (PRE, 2011-2012), transition (TRANS, 2013-2014), and after introduction of L4 (POST, 2015-2017). Analytical section: variables associated with a diagnosis of leptospirosis were investigated in a case-control study using multivariable logistic regression, and focusing on vaccination. RESULTS: The number of dogs diagnosed with leptospirosis (AKI-L) decreased from 56.5 (PRE) to 15.7 (POST) cases/year while controls increased from 16.5 to 38.0 cases/year. Control dogs (AKI-nL) showed a decrease in L2 vaccination (100% to 26%) and an increase in L4 vaccination (0% to 70%). The odds ratio for vaccinated dogs to be diagnosed with leptospirosis was 0.11 (95% confidence interval [CI], 0.06-0.22; P < .001) for L4 and 2.08 (0.58-7.42; P = .26) for L2. CONCLUSIONS AND CLINICAL IMPORTANCE: The introduction of L4 was associated with a marked decrease in dogs with leptospirosis and AKI in Switzerland. Use of the L4 vaccine was associated with significantly decreased odds of disease.

First European report of Francisella tularensis subsp. holarctica isolation from a domestic cat.

Francisella tularensis subsp. holarctica is a select agent causing life-threatening tularemia. It has been isolated from humans and animals, mainly lagomorphs and rodents, rarely other wild carnivore species. Increasing numbers of human tularemia cases have been reported during the last 5 years in Switzerland. Here we report the first isolation of Francisella tularensis subsp. holarctica from a domestic cat in Europe and compare its genome sequence with other Swiss isolates. The cat isolate shows a close phylogenetic relationship with a contemporary hare isolate from close geographic proximity, indicating a possible epidemiological link.

Toxicosis with grapes or raisins causing acute kidney injury and neurological signs in dogs.

BACKGROUND: The ingestion of grapes or raisins has been reported to cause acute kidney injury (AKI) in dogs, with a clinical picture dominated by early gastrointestinal signs and rapidly developing uremia. Ataxia is mentioned in a few reports, but not further characterized. OBJECTIVES: To evaluate the clinical, laboratory, and pathological features of dogs diagnosed with grape or raisin toxicosis (GRT) with emphasis on renal and neurological manifestations, compared to a control group of dogs with AKI from other causes. ANIMALS: Fifteen client-owned dogs with GRT and 74 control dogs. METHODS: Retrospective study over 17 months. RESULTS: All dogs with GRT were presented with severe AKI (grade 4, n = 5; grade 5, n = 10). Eleven dogs (73%) had marked forebrain, cerebellar, or vestibular signs. These manifestations dominated the clinical picture in some dogs, but were not associated with the severity of azotemia or the presence of systemic hypertension. Eight dogs (53%) survived, and 5 dogs experienced a complete neurological recovery. Causes of death were unrelated to the neurological manifestations. Neuropathological examination of 4 dogs did not identify any structural central nervous system abnormality. Only 2 control dogs (3%) displayed neurological signs with seizures unrelated to the AKI; 42 control dogs (57%) survived. CONCLUSIONS AND CLINICAL IMPORTANCE: Severe forebrain, cerebellar, or vestibular signs may be an important feature of GRT and dominate the early clinical picture. The described features suggest a reversible functional brain injury specific to GRT and unrelated to uremia.

Evaluation of a novel non-surgical post-pyloric feeding technique in dogs with severe acute kidney injury.

OBJECTIVES: To evaluate the placement and the performance of a new post-pyloric feeding (PPF) system, and compare it to esophageal (E) feeding in dogs with severe acute kidney injury (AKI). DESIGN: Prospective study. SETTING: University teaching hospital. ANIMALS: Twenty client-owned dogs with severe AKI treated with hemodialysis. INTERVENTIONS: Dogs were randomly assigned to 2 feeding techniques, PPF or E feeding; for PPF, an 8-Fr jejunal (J) tube was introduced through a standard 18-Fr E-tube and advanced endoscopically through the pylorus. A commercial liquid diet was administered at a continuous rate for at least 5 days. After the removal of the internal J-tube, feeding was continued through the E-tube. Control dogs were fed blended food 5 times daily through an E-tube. Placement technique, nutritional efficiency, and metabolic response to nutritional support were compared between the groups. MEASUREMENTS AND MAIN RESULTS: Feeding tubes were placed a medium of 1 day (interquartile range, 0.8 d) after presentation. Median procedure-time in the group PPF was 26.5 min (11.5 min), including 8.5 min (2.7 min) for the E-tube and 17.5 min (5.0 min) for the J-tube. J-tubes were used for 5 days (1.5 d) with oral displacement observed in 3/10 dogs. The goal to administer 130% resting energy requirement within 3 days was reached in 13/20 dogs (8/10 PPF, 5/10 E). Despite nutritional support, dogs lost 1.2% body weight per day (1.1%), with no difference between the groups. The metabolic improvement was comparable between the groups, despite marked differences in the diet composition. CONCLUSIONS: This new feeding technique can be recommended as an efficient method for early nutritional support in dogs with severe AKI. The protein-energy wasting observed despite active nutritional support suggests, however, the necessity for increased feeding targets or qualitative changes in diet composition for dogs with AKI.

Acute kidney injury due to Leptospira interrogans in 4 foals and use of renal replacement therapy with intermittent hemodiafiltration in 1 foal.

Four 2-month-old foals were presented to an equine hospital with acute kidney injury caused by Leptospira interrogans infection. Clinical signs were nonspecific and included lethargy, fever, and unwillingness to nurse. The most important hematologic and clinicopathologic findings were azotemia, anemia, thrombocytopenia, hyponatremia, and hypochloremia. The diagnosis was based on urinary real-time PCR, serology using a microscopic agglutination test, or both. The most important serovars involved were L. interrogans serogroup australis serovar Bratislava and Australis. Treatment consisted of IV fluid therapy and antimicrobial treatment. Renal replacement therapy with hemodiafiltration was performed in 1 of the foals. All foals survived to discharge. This report highlights the importance of early diagnosis and treatment in foals with acute kidney injury caused by L. interrogans infection.

Membrane-based therapeutic plasma exchange in dogs: Prescription, anticoagulation, and metabolic response.

BACKGROUND: Therapeutic plasma exchange (TPE) is used increasingly in small animals to remove circulating large molecular products such as antibodies, pathogenic proteins, and protein-bound toxins. Specific, efficient, and safe protocols need to be developed. HYPOTHESIS/OBJECTIVES: To describe the technique of membrane-based TPE, the resulting physiological and metabolic changes, and to define an adequate regional citrate anticoagulation protocol. ANIMALS: Thirty-four dogs treated with TPE (2011-2017). METHODS: Retrospective review of all TPE treatments performed at the Vetsuisse Faculty, University of Bern, identified through a search of the institutional database for extracorporeal treatments. RESULTS: Sixty-four treatments were performed, resulting in 1.0 plasma volume exchange (range, 0.4-1.1). Replacement fluids included fresh frozen plasma (12%-100% volume), colloids (0%-52%), human albumin (0%-41%), and saline (0%-70%). Anticoagulation was performed with regional citrate (n = 24), systemic heparinization (n = 2), or combined (n = 38). Main relevant laboratory changes included a 24.7% decrease in total proteins (interquartile range, 16.7-31.4; P < .001), 53% in fibrinogen (-30 to 63; P = .009), 36% in bilirubin (13-43, P = .02), 9.0% in urea (0.7-15.7; P < .001), and 4.5% in creatinine (-6.6 to 10.6; P = .006). Citrate accumulation was evidenced in all dogs, more pronounced in those with renal but not with hepatic impairment. Maximal tolerable citrate rates were estimated as 5.5 and 9.0 µmol/kg/min for treatments in dogs with and without renal impairment, respectively. Complications were observed in 22 treatments (34%) and were fatal in 2 dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Therapeutic plasma exchange causes metabolic and biochemical alterations. Understanding these effects makes possible to anticipate most complications and to improve safety of the procedure.

Canine leptospirosis in Switzerland-A prospective cross-sectional study examining seroprevalence, risk factors and urinary shedding of pathogenic leptospires.

Leptospirosis is an important worldwide zoonosis. While human leptospirosis remains rare in Switzerland, the incidence of canine leptospirosis is unusually high compared to other European countries. The aims of this cross-sectional study were to determine the exposure of asymtomatic dogs to pathogenic Leptospira in Switzerland, to characterise risk factors associated with seropositivity and to determine the prevalence of urinary shedding. Sampling was stratified to cover the whole of Switzerland. Sera were tested by microscopic agglutination test for antibodies against a panel of 12 serovars. Urine was tested for pathogenic Leptospira using a LipL32 real-time PCR. Of 377 sera, 55.7% (95%CI 51.2-60.7) showed a reciprocal MAT titre of ≥1:40 and 24.9% (95%CI 20.7-29.4) of ≥1:100 to at least one serovar. Seropositivity (MAT ≥1:100) was most common to serovars Australis (14.9%; 95% CI 11.4-18.6) and Bratislava (8.8%; 95%CI 6.1-11.7), followed by Copenhageni (6.1%; 95%CI 3.7-8.5), Canicola (5%; 95%CI 2.9-7.4), Grippotyphosa (4.5%; 95%CI 2.7-6.9), Pomona (4%; 95%CI 2.1-6.1), Autumnalis (2.7%; 95%CI 1.3-4.2) and Icterohaemorrhagiae (1.6%; 95%CI 0.5-2.9). In unvaccinated dogs (n=84) the prevalence of a MAT titre ≥100 was 17.9% (95%CI 10.7-26.2), with a similar distribution of reactive serovars. Variables associated with seropositivity (≥1:40) to any serovar included age (OR 1.29/year; 95%CI: 1.1-1.5) and bioregion with higher risks in the regions Northern Alps (OR 14.5; 95%CI 2.2-292.7), Central Plateau (OR 12.3; 95%CI 2.0-244.1) and Jura (OR 11.2; 95%CI 1.7-226.7) compared to Southern Central Alps. Dogs living with horses were significantly more likely to have antibodies to serovar Bratislava (OR 4.68;95%CI 1.2-17.2). Hunting was a significant risk factor for seropositivtiy to serovar Grippotyphosa (OR 8.03; 95%CI 1.6-30.8). Urine qPCR positivity was uncommon (1/408 dogs; 0.2%; 95% CI0-0.7). These results demonstrate that dogs in Switzerland are commonly exposed to pathogenic Leptospira; however, the risk of dogs contributing to the spread of Leptospira in the environment appears low.

Assessment of the expression of biomarkers of uremic inflammation in dogs with renal disease.

OBJECTIVE: To assess the expression of inflammatory cytokines and enzymes in venous whole blood of dogs with impaired renal function attributable to various causes. ANIMALS: 46 dogs with acute kidney injury (AKI), 8 dogs with chronic kidney disease (CKD), and 10 healthy dogs. PROCEDURES: Dogs with AKI and CKD were prospectively enrolled during 2010 if they met inclusion criteria. Demographic and laboratory characteristics were evaluated for each dog, and expression of inflammatory cytokines (interleukin [IL]-1α, IL-1ß, IL-8, tumor necrosis factor [TNF]-α, IL-10, and transforming growth factor [TGF]-ß) and enzymes (inducible nitric oxide synthase [iNOS] and 5-lipoxygenase [5-LO]) was measured in venous whole blood obtained at initial evaluation. RESULTS: Dogs with impaired renal function had markedly higher expression of the cytokines IL-1α, IL-1ß, and TGF-ß and the enzyme 5-LO, compared with expression in healthy dogs. Additionally, 17 of 46 AKI dogs (but none of the CKD dogs) had higher IL-8 mRNA expression and 3 of 8 CKD dogs (but only 2/46 AKI dogs) had higher TNF-α expression, compared with results for healthy dogs. No significant difference between renal disease groups was detected for inflammatory markers and laboratory variables, degree of azotemia, or cause of impaired renal function. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, expression of the cytokines IL-1α, IL-1ß, and TGF-ß and the enzyme 5-LO was clearly increased in dogs with renal disease, which suggested that these markers were part of an inflammatory response in animals with AKI or CKD.

Expression Profile of Cytokines and Enzymes mRNA in Blood Leukocytes of Dogs with Leptospirosis and Its Associated Pulmonary Hemorrhage Syndrome.

BACKGROUND: Dogs with leptospirosis show similar organ manifestations and disease course as human patients, including acute kidney injury and pulmonary hemorrhage, making this naturally-occurring infection a good animal model for human leptospirosis. Expression patterns of cytokines and enzymes have been correlated with disease manifestations and clinical outcome in humans and animals. The aim of this study was to describe mRNA expression of pro- and anti-inflammatory mediators in canine leptospirosis and to compare it with other renal diseases to identify patterns characterizing the disease and especially its pulmonary form. METHODOLOGY AND PRINCIPAL FINDINGS: The mRNA abundance of cytokines (IL-1α, IL-1ß, IL-8, IL-10, TNF-α, TGF-ß) and enzymes (5-LO, iNOS) was measured prospectively in blood leukocytes from 34 dogs with severe leptospirosis and acute kidney injury, including 22 dogs with leptospirosis-associated pulmonary hemorrhages. Dogs with leptospirosis were compared to 14 dogs with acute kidney injury of other origin than leptospirosis, 8 dogs with chronic kidney disease, and 10 healthy control dogs. Canine leptospirosis was characterized by high 5-LO and low TNF-α expression compared to other causes of acute kidney injury, although the decreased TNF-α expression was also seen in chronic kidney disease. Leptospirosis-associated pulmonary hemorrhage was not characterized by a specific pattern, with only mild changes noted, including increased IL-10 and decreased 5-LO expression on some days in affected dogs. Fatal outcome from pulmonary hemorrhages was associated with low TNF-α, high IL-1ß, and high iNOS expression, a pattern possibly expressed also in dogs with other forms of acute kidney injury. CONCLUSION: The patterns of cytokine and enzyme expression observed in the present study indicate a complex pro- and anti-inflammatory response to the infection with leptospires. The recognition of these signatures may be of diagnostic and prognostic relevance for affected individuals and they may indicate options for newer therapies targeting the identified pathways.

Comparison of the in vitro effects of saline, hypertonic hydroxyethyl starch, hypertonic saline, and two forms of hydroxyethyl starch on whole blood coagulation and platelet function in dogs.

OBJECTIVE: To compare the in vitro effects of hypertonic solutions and colloids to saline on coagulation in dogs. DESIGN: In vitro experimental study. SETTING: Veterinary teaching hospital. ANIMALS: Twenty-one adult dogs. INTERVENTIONS: Blood samples were diluted with saline, 7.2% hypertonic saline solution with 6% hydroxyethylstarch with an average molecular weight of 200 kDa and a molar substitution of 0.4 (HH), 7.2% hypertonic saline (HTS), hydroxyethyl starch (HES) 130/0.4 or hydroxyethyl starch 600/0.75 at ratios of 1:22 and 1:9, and with saline and HES at a ratio of 1:3. MEASUREMENTS AND MAIN RESULTS: Whole blood coagulation was analyzed using rotational thromboelastometry (extrinsic thromboelastometry-cloting time (ExTEM-CT), maximal clot firmness (MCF) and clot formation time (CFT) and fibrinogen function TEM-CT (FibTEM-CT) and MCF) and platelet function was analyzed using a platelet function analyzer (closure time, CTPFA ). All parameters measured were impaired by saline dilution. The CTPFA was prolonged by 7.2% hypertonic saline solution with 6% hydroxyethylstarch with an average molecular weight of 200 kDa and a molar substitution of 0.4 (HH) and HTS but not by HES solutions. At clinical dilutions equivalent to those generally administered for shock (saline 1:3, HES 1:9, and hypertonic solutions 1:22), CTPFA was more prolonged by HH and HTS than other solutions but more by saline than HES. No difference was found between the HES solutions or the hypertonic solutions. ExTEM-CFT and MCF were impaired by HH and HTS but only mildly by HES solutions. At clinically relevant dilutions, no difference was found in ExTEM-CFT between HTS and saline or in ExTEM-MCF between HH and saline. No consistent difference was found between the 2 HES solutions but HH impaired ExTEM-CFT and MCF more than HTS. At high dilutions, FibTEM-CT and -MCF and ExTEM-CT were impaired by HES. CONCLUSIONS: Hypertonic solutions affect platelet function and whole blood coagulation to a greater extent than saline and HES. At clinically relevant dilutions, only CTPFA was markedly more affected by hypertonic solutions than by saline. At high dilutions, HES significantly affects coagulation but to no greater extent than saline at clinically relevant dilutions.

Immunohistochemical detection of IgM and IgG in lung tissue of dogs with leptospiral pulmonary haemorrhage syndrome (LPHS).

Leptospiral pulmonary haemorrhage syndrome (LPHS) is a severe form of leptospirosis. Pathogenic mechanisms are poorly understood. Lung tissues from 26 dogs with LPHS, 5 dogs with pulmonary haemorrhage due to other causes and 6 healthy lungs were labelled for IgG (n=26), IgM (n=25) and leptospiral antigens (n=26). Three general staining patterns for IgG/IgM were observed in lungs of dogs with LPHS with most tissues showing more than one staining pattern: (1) alveolar septal wall staining, (2) staining favouring alveolar surfaces and (3) staining of intra-alveolar fluid. Healthy control lung showed no staining, whereas haemorrhagic lung from dogs not infected with Leptospira showed staining of intra-alveolar fluid and occasionally alveolar septa. Leptospiral antigens were not detected. We conclude that deposition of IgG/IgM is demonstrable in the majority of canine lungs with naturally occurring LPHS, similar to what has been described in other species. Our findings suggest involvement of the host humoral immunity in the pathogenesis of LPHS and provide further evidence to support the dog as a natural disease model for human LPHS.

Increasing incidence of canine leptospirosis in Switzerland.

A marked increase in canine leptospirosis was observed in Switzerland over 10 years with a peak incidence of 28.1 diagnosed cases/100,000 dogs/year in the most affected canton. With 95% affected dogs living at altitudes <800 m, the disease presented a seasonal pattern associated with temperature (r2 0.73) and rainfall (r2 0.39), >90% cases being diagnosed between May and October. The increasing yearly incidence however was only weakly correlated with climatic data including number of summer (r2 0.25) or rainy days (r2 0.38). Serovars Australis and Bratislava showed the highest seropositivity rates with 70.5% and 69.1%, respectively. Main clinical manifestations included renal (99.6%), pulmonary (76.7%), hepatic (26.0%), and hemorrhagic syndromes (18.2%), leading to a high mortality rate (43.3%). Similar to the human disease, liver involvement had the strongest association with negative outcome (OR 16.3). Based on these data, canine leptospirosis presents similar features and severity as the human infection for which it therefore can be considered a model. Its re-emergence in a temperate country with very high incidence rates in canines should thus be viewed as a warning and emphasize the need for increased awareness in other species.

Evaluation of the diagnostic value of serologic microagglutination testing and a polymerase chain reaction assay for diagnosis of acute leptospirosis in dogs in a referral center.

OBJECTIVE: To determine the diagnostic value of a serologic microagglutination test (MAT) and a PCR assay on urine and blood for the diagnosis of leptospirosis in dogs with acute kidney injury (AKI). DESIGN: Cross-sectional study. Animals-76 dogs with AKI in a referral hospital (2008 to 2009). PROCEDURES: Dogs' leptospirosis status was defined with a paired serologic MAT against a panel of 11 Leptospira serovars as leptospirosis-associated (n = 30) or nonleptospirosis-associated AKI (12). In 34 dogs, convalescent serologic testing was not possible, and leptospirosis status was classified as undetermined. The diagnostic value of the MAT single acute or convalescent blood sample was determined in dogs in which leptospirosis status could be classified. The diagnostic value of a commercially available genus-specific PCR assay was evaluated by use of 36 blood samples and 20 urine samples. RESULTS: Serologic acute testing of an acute blood sample had a specificity of 100% (95% CI, 76% to 100%), a sensitivity of 50% (33% to 67%), and an accuracy of 64% (49% to 77%). Serologic testing of a convalescent blood sample had a specificity of 92% (65% to 99%), a sensitivity of 100% (87% to 100%), and an accuracy of 98% (88% to 100%). Results of the Leptospira PCR assay were negative for all samples from dogs for which leptospirosis status could be classified. CONCLUSIONS AND CLINICAL RELEVANCE: Serologic MAT results were highly accurate for diagnosis of leptospirosis in dogs, despite a low sensitivity for early diagnosis. In this referral setting of dogs pretreated with antimicrobials, testing of blood and urine samples with a commercially available genus-specific PCR assay did not improve early diagnosis.